U.S. patent application number 12/615104 was filed with the patent office on 2010-03-18 for nucleic acid molecules and other molecules associated with the methionine synthesis and degradation pathways.
Invention is credited to Stefan A. Bledig, Joseph R. Byrum, Jingdong Liu.
Application Number | 20100071097 12/615104 |
Document ID | / |
Family ID | 46328254 |
Filed Date | 2010-03-18 |
United States Patent
Application |
20100071097 |
Kind Code |
A1 |
Bledig; Stefan A. ; et
al. |
March 18, 2010 |
Nucleic Acid Molecules and Other Molecules Associated with the
Methionine Synthesis and Degradation Pathways
Abstract
The present invention is in the field of plant biochemistry.
More specifically the invention relates to nucleic acid sequences
from plant cells, in particular, DNA sequences from maize and
soybean plants associated with the methionine pathway. The
invention encompasses nucleic acid molecules that encode proteins
and fragments of proteins. In addition, the invention also
encompasses proteins and fragments of proteins so encoded and
antibodies capable of binding these proteins or fragments. The
invention also relates to methods of using the nucleic acid
molecules, proteins and fragments of proteins and antibodies, for
example for genome mapping, gene identification and analysis, plant
breeding, preparation of constructs for use in plant gene
expression and transgenic plants.
Inventors: |
Bledig; Stefan A.;
(Chesterfield, MO) ; Byrum; Joseph R.; (Des
Moines, IA) ; Liu; Jingdong; (Ballwin, MO) |
Correspondence
Address: |
ARNOLD & PORTER LLP
555 TWELFTH STREET, N.W., ATTN: IP DOCKETING
WASHINGTON
DC
20004
US
|
Family ID: |
46328254 |
Appl. No.: |
12/615104 |
Filed: |
November 9, 2009 |
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Current U.S.
Class: |
800/298 ;
47/58.1SE; 536/23.6 |
Current CPC
Class: |
C12N 9/88 20130101 |
Class at
Publication: |
800/298 ;
536/23.6; 47/58.1SE |
International
Class: |
A01H 5/00 20060101
A01H005/00; A01H 5/10 20060101 A01H005/10; C07H 21/04 20060101
C07H021/04; A01H 3/00 20060101 A01H003/00 |
Claims
1-11. (canceled)
12. A transformed plant comprising a nucleic acid molecule which
comprises: (a) an exogenous promoter region which functions in a
plant cell to cause the production of an mRNA molecule; which is
linked to; (b) a structural nucleic acid molecule, wherein said
structural nucleic acid molecule comprises a nucleic acid sequence,
wherein said nucleic acid sequence shares between 100% and 90%
sequence identity with a nucleic acid sequence selected from the
group consisting of SEQ ID NO: 1 through SEQ ID NO: 3204 and
complements thereof, or which is linked to (c) a 3' non-translated
sequence that functions in said plant cell to cause the termination
of transcription and the addition of polyadenylated ribonucleotides
to said 3' end of said mRNA molecule.
13. The transformed plant according to claim 12, wherein said
nucleic acid sequence is the complement of a nucleic acid sequence
selected from the group consisting of SEQ ID NO: 1 through SEQ ID
NO: 3204.
14. The transformed plant according to claim 12, wherein said
nucleic acid sequence is in the antisense orientation of a nucleic
acid sequence selected from the group consisting of SEQ ID NO: 1
through SEQ ID NO: 3204.
15. The transformed plant according to claim 12, wherein said
nucleic acid sequence shares between 100% and 95% sequence identity
with a nucleic acid sequence selected from the group consisting of
SEQ ID NO: 1 through SEQ ID NO: 3204 and complements thereof.
16. The transformed plant according to claim 15, wherein said
nucleic acid sequence shares between 100% and 98% sequence identity
with a nucleic acid sequence selected from the group consisting of
SEQ ID NO: 1 through SEQ ID NO: 3204 and complements thereof.
17. The transformed plant according to claim 16, wherein said
nucleic acid sequence shares between 100% and 99% sequence identity
with a nucleic acid sequence selected from the group consisting of
SEQ ID NO: 1 through SEQ ID NO: 3204 and complements thereof.
18. The transformed plant according to claim 17, wherein said
nucleic acid sequence shares 100% sequence identity with a nucleic
acid sequence selected from the group consisting of SEQ ID NO: 1
through SEQ ID NO: 3204 and complements thereof.
19. A transformed seed comprising a transformed plant cell
comprising a nucleic acid molecule which comprises: (a) an
exogenous promoter region which functions in said plant cell to
cause the production of an mRNA molecule; which is linked to; (b) a
structural nucleic acid molecule, wherein said structural nucleic
acid molecule comprises a nucleic acid sequence, wherein said
nucleic acid sequence shares between 100% and 90% sequence identity
with a nucleic acid sequence selected from the group consisting of
SEQ ID NO: 1 through SEQ ID NO: 3204 and complements thereof, which
is linked to (c) a 3' non-translated sequence that functions in
said plant cell to cause the termination of transcription and the
addition of polyadenylated ribonucleotides to said 3' end of said
mRNA molecule.
20. The transformed seed according to claim 19, wherein said
nucleic acid sequence is the complement of a nucleic acid sequence
selected from the group consisting of SEQ ID NO: 1 through SEQ ID
NO: 3204.
21. The transformed seed according to claim 20, wherein said
exogenous promoter region functions in a seed cell.
22. The transformed seed according to claim 21, wherein said
nucleic acid sequence shares between 100% and 95% sequence identity
with a nucleic acid sequence selected from the group consisting of
SEQ ID NO: 1 through SEQ ID NO: 3204 and complements thereof.
23. The transformed seed according to claim 22, wherein said
nucleic acid sequence shares between 100% and 98% sequence identity
with a nucleic acid sequence selected from the group consisting of
SEQ ID NO: 1 through SEQ ID NO: 3204 and complements thereof.
24. The transformed seed according to claim 23, wherein said
nucleic acid sequence shares between 100% and 99% sequence identity
with a nucleic acid sequence selected from the group consisting of
SEQ ID NO: 1 through SEQ ID NO: 3204 and complements thereof.
25. The transformed seed according to claim 24, wherein said
nucleic acid sequence shares 100% sequence identity with a nucleic
acid sequence selected from the group consisting of SEQ ID NO: 1
through SEQ ID NO: 3204 and complements thereof.
26. A method of growing a transgenic plant comprising (a) planting
a transformed seed comprising a nucleic acid sequence, wherein said
nucleic acid sequence shares between 100% and 90% sequence identity
with a nucleic acid sequence selected from the group consisting of
SEQ ID NO: 1 through SEQ ID NO: 3204 and complements thereof, and
(b) growing a plant from said seed.
27. A substantially purified nucleic acid molecule comprising a
nucleic acid sequence, wherein said nucleic acid sequence shares
between 100% and 90% sequence identity with a nucleic acid sequence
selected from the group consisting of SEQ ID NO: 1 through SEQ ID
NO: 3204 and complements thereof.
28. The substantially purified nucleic acid molecule of claim 27,
wherein said nucleic acid molecule encodes a maize protein or
fragment thereof.
29. The substantially purified nucleic acid molecule of claim 28,
wherein said nucleic acid molecule encodes a soybean protein or
fragment thereof.
Description
CROSS REFERENCE TO RELATED APPLICATIONS
[0001] This application claims the benefit of applications No.
60/067,000 filed Nov. 24, 1997, No. 60/066,873 filed Nov. 25, 1997,
No. 60/069,472 filed Dec. 9, 1997, No. 60/074,201 filed Feb. 10,
1998, No. 60/074,282 filed Feb. 10, 1998, No. 60/074,280 filed Feb.
10, 1998, No. 60/074,281 filed Feb. 10, 1998, No. 60/074,566 filed
Feb. 12, 1998, No. 60/074,567 filed Feb. 12, 1998, No. 60/074,565
filed Feb. 12, 1998, No. 60/075,462 filed Feb. 19, 1998, No.
60/074,789 filed Feb. 19, 1998, No. 60/075,459 filed Feb. 19, 1998,
No. 60/075,461 filed Feb. 19, 1998, No. 60/075,464 filed Feb. 19,
1998, No. 60/075,460 filed Feb. 19, 1998, No. 60/075,463 filed Feb.
19, 1998, No. 60/077,231 filed Mar. 9, 1998, No. 60/077,229 filed
Mar. 9, 1998, No. 60/077,230 filed Mar. 9, 1998, No. 60/078,031
filed Mar. 16, 1998, No. 60/078,368 filed Mar. 18, 1998, No.
60/080,844 filed Apr. 7, 1998, No. 60/083,067 filed Apr. 27, 1998,
"Nucleic Acid Molecules and Other Molecules Associated with Plants"
docket No. 38-21(15348)A filed Apr. 29, 1998, No. 60/083,387 filed
Apr. 29, 1998, No. 60/083,388 filed Apr. 29, 1998, No. 60/083,389
filed Apr. 29, 1998, "Nucleic Acid Molecules and Other Molecules
Associated with the Ethylene Biosynthetic Pathway" docket No.
04983.0018/38-21(15097)A filed May 8, 1998, No. 60/085,245 filed
May 13, 1998, No. 60/085,224 filed May 13, 1998, No. 60/085,223
filed May 13, 1998, No. 60/085,222 filed May 13, 1998, No.
60/086,186 filed May 21, 1998, No. 60/086,339 filed May 21, 1998,
No. 60/086,187 filed May 21, 1998, No. 60/086,185 filed May 21,
1998, No. 60/086,184 filed May 21, 1998, No. 60/086,183 filed May
21, 1998, No. 60/086,188 filed May 21, 1998, No. 60/089,524 filed
Jun. 16, 1998, No. 60/089,810 filed Jun. 18, 1998, No. 60/089,814
filed Jun. 18, 1998, "Nucleic acid molecules and other molecules
associated with the Plant Sugar and Nitrogen Transporters Pathway"
docket No. 04983.0043/38-21(15412)A filed Jun. 30, 1998, No.
60/092,036 filed Jul. 8, 1998, No. 60/099,667 filed Sep. 9, 1998,
No. 60/099,668 filed Sep. 9, 1998, No. 60/099,670 filed Sep. 9,
1998, No. 60/099,697 filed Sep. 9, 1998, No. 60/100,674 filed Sep.
16, 1998, No. 60/100,673 filed Sep. 16, 1998, No. 60/100,672 filed
Sep. 16, 1998, No. 60/101,132 filed Sep. 21, 1998, No. 60/101,130
filed Sep. 21, 1998, "Nucleic acid molecules and other molecules
associated with Plants" docket No. 38-21(15459)A filed Sep. 21,
1998, No. 60/101,344 filed Sep. 22, 1998, No. 60/101,347 filed Sep.
22, 1998, No. 60/101,343 filed Sep. 22, 1998, No. 60/104,126 filed
Oct. 13, 1998, No. 60/104,128 filed Oct. 13, 1998, No. 60/104,127
filed Oct. 13, 1998, No. 60/104,124 filed Oct. 13, 1998, "Nucleic
Acid Molecules and Other Molecules Associated with Plants" docket
No. 38-21(15445)A filed Nov. 18, 1998 and "Nucleic Acid Molecules
and other Molecules associated with Plants" docket No. 38-21(15592)
filed Nov. 18, 1998 hereby incorporated by reference herein in
their entirety.
FIELD OF THE INVENTION
[0002] The present invention is in the field of plant biochemistry.
More specifically the invention relates to nucleic acid sequences
from plant cells, in particular, DNA sequences from maize and
soybean plants associated with the methionine pathway. The
invention encompasses nucleic acid molecules that encode proteins
and fragments of proteins. In addition, the invention also
encompasses proteins and fragments of proteins so encoded and
antibodies capable of binding these proteins or fragments. The
invention also relates to methods of using the nucleic acid
molecules, proteins and fragments of proteins and antibodies, for
example for genome mapping, gene identification and analysis, plant
breeding, preparation of constructs for use in plant gene
expression and transgenic plants.
BACKGROUND OF THE INVENTION
I. Methoinine Synthesis Pathway
[0003] The amino acid, L-methionine, is synthesized in higher
plants via a pathway that starts with L-aspartate. This pathway has
been studied (Azevedo et al., Phytochemistry 46:395-419 (1997), the
entirety of which is herein incorporated by reference).
L-methionine is one of four so-called aspartate-derived amino acids
(along with L-lysine, L-threonine and L-isoleucine)(Miflin et al.,
In: Nitrogen Assimilation in Plants, Hewitt et al., (eds.),
Academic Press, New York, 335 (1997); Bryan, In: The Biochemistry
of Plants, Miflin (ed.), Academic Press, New York, 403 (1980); Lea
et al., In: The Chemistry and Biochemistry of Amino Acids, Barrett
et al., (eds.), London, 5:197 (1985); Bryan, In: The Biochemistry
of Plants, Miflin et al., (eds.), Academic Press, San Diego, 16:161
(1990), all of which are herein incorporated by reference in their
entirety).
[0004] The methionine-specific part of the aspartate pathway
includes the following enzymes: aspartate kinase (EC 2.7.2.4),
aspartate-semialdehyde dehydrogenase (EC 1.2.1.11), homoserine
dehydrogenase (EC 1.1.1.3), homoserine kinase (EC 2.7.1.39),
cystathionine .gamma.-synthase (EC 4.2.99.9), cystathionine
.beta.-lyase (EC 4.4.1.8) and methionine synthase (EC
2.1.1.14).
[0005] Aspartate kinase catalyzes the first reaction of the pathway
in which aspartate is converted to .beta.-aspartyl phosphate. This
enzyme has been isolated and characterized from plant sources
including maize, barley, carrot, pea and soybean. These studies
have revealed that there are multiple isoenzymes of aspartate
kinase and the isoenzymes differ with respect to both feedback
inhibition sensitivity and expression profile (tissue and
developmental stage). Feedback inhibition is mediated by lysine and
threonine. Transgenic plants which express an unregulated aspartate
kinase have demonstrated increased flux through the aspartate
pathway. Pathway regulation is reported to be exerted, at least in
part, via control of this enzyme's activity.
[0006] Aspartate semialdehyde dehydrogenase catalyses the second
pathway reaction and converts .beta.-aspartyl phosphate to
aspartate semialdehyde via an NADPH-dependent reaction. Gengenbach
et al., Crop Science 18:472-476 (1978), the entirety of which is
herein incorporated by reference, report the isolation of aspartate
semialdehyde dehydrogenase from maize suspension culture cells.
These suspension cultures did not exhibit feedback inhibition of
the enzyme in the presence of aspartate-derived amino acids, with
the exception of methionine, for which some feedback sensitivity
was observed. Aspartate semialdehyde dehydrogenase enzyme activity
has been reported in maize shoot, maize root and maize kernel
(Gengenbach et al., Crop Science 18:472-476 (1978)).
[0007] Homoserine dehydrogenase catalyzes the next step of the
pathway in which homoserine is generated from aspartate
semialdehyde in a reaction requiring NADH or NADPH. Homoserine
dehydrogenase enzyme has been studied in higher plants and multiple
isoenzyme forms have been reported (Bryan et al., Biochemistry and
Biophysics Research Communications 41:1211-1217 (1970); Gengenbach
et al., Crop Science 18:472-476 (1978); Dotson et al., Plant
Physiology 91:1602-1608 (1989); Dotson et al., Plant Physiology
93:98-104 (1989); Azevedo et al., Phytochemistry 31:3725-3730
(1992); Azevedo et al., Phytochemistry 31:3731-3734 (1992);
Brennecke et al., Phytochemistry 41:707 (1996); Aarnes, Plant
Science Letters 9:137-145 (1977); Bright et al., Biochemical
Genetics 200:229-243 (1982); Aruda et al., Plant Physiology
76:442-446 (1984); Lea et al., In: Barley: Genetics, Molecular
Biology and Biotechnology Shewrey (ed.), CAB International, Oxford
181 (1992); Davies et al., Plant Science Letters 9:323-332 (1977);
Davies et al., Plant Physiology 62:536-541 (1978); Matthews et al.,
Zeitschrifi fur Naturforschung, Section Bioscience 34:1177-1185
(1979); Relton et al., Biochimica et Biophysica Acta 953:48-60
(1988); Aarnes et al., Phytochemistry 13:2717-2724 (1974); Lea et
al., FEBS Letters 98:165-168 (1979); Matthews et al., Canadian
Journal of Botany 57:299-304 (1979), all of which references are
incorporated herein in their entirety). The isoenzymes have been
found to differ with respect to sensitivity to threonine-mediated
feedback inhibition, with both sensitive and insensitive forms
being isolated from maize suspension cultures and seedlings (Miflin
et al., In: Nitrogen Assimilation of Plants, Hewitt et al., (eds.),
Academic Press, New York, 335 (1997); Bryan, In: The Biochemistry
of Plants, Miflin (ed.), Academic Press, New York, 5:403
(1980)).
[0008] There is evidence that plants also possess a bifunctional
enzyme with both aspartate kinase and homoserine dehydrogenase
activities (Lea et al., In: The Chemistry and Biochemistry of Amino
Acids, Barrett et al. (eds), London, 5:197 (1985), the entirety of
which is herein incorporated by reference; Bryan, In: The
Biochemistry of Plants, Miflin (ed.), Academic Press, New York,
5:161 (1990), the entirety of which is herein incorporated by
reference). Clones of these bifunctional enzymes have been isolated
from Arabidopsis thaliana (Giovanelli et al., In: The Biochemistry
of Plants, Miflin (ed.), Academic Press, New York 453 (1990), the
entirety of which is herein incorporated by reference) carrot
(Giovanelli et al., Plant Physiology 90:1584-1599 (1989), the
entirety of which is herein incorporated by reference), maize
(Singh et al., Amino Acids 7:165-168 (1994), the entirety of which
is herein incorporated by reference) and soybean (Matthews et al.,
In: Biosynthesis and Molecular Regulation of Amino Acids in Plants,
p 294, Singh et al. (eds.), American Society of Plant
Physiologists, Rockville, Md. (1992), the entirety of which is
herein incorporated by reference).
[0009] The next reported enzymatic step leading to methionine
biosynthesis in higher plants is the final common reaction shared
by other amino acid end products (threonine and isoleucine). The
reaction is catalyzed by homoserine kinase and it generates
O-phosphohomoserine from homoserine, with ATP serving as the
phosphate donor. Exceptions are Pisum sativum and Lathyrus sitivus
which synthesize O-acetylhomoserine and O-oxalylhomoserine,
respectively (Thomas and Surdin-Kerjan, Microbiol. Mol. Biol. Rev.
61:503-532 (1997), the entirety of which is herein incorporated by
reference). Enteric bacteria use O-succinylhomoserine, while
several gram-positive bacteria, yeasts and fungi use
O-acetylhomoserine (formed using homoserine O-acetyltransferase (EC
2.3.1.31) (Thomas and Surdin-Kerjan, Microbiol. Mol. Biol. Rev.
61:503-532 (1997)). Homoserine kinase has been reported from
multiple higher plant sources (Galili, The Plant Cell 7:899-906
(1995), the entirety of which is herein incorporated by reference;
Rees et al., Biochemical Journal 309:999-1107 (1995), the entirety
of which is herein incorporated by reference; Bryan et al.,
Biochemistry and Biophysics Research Communications 41:1211-1217
(1970), the entirety of which is herein incorporated by reference;
Gengenbach et al., Crop Science 18:472-476 (1978), Dotson et al.,
Plant Physiology 91:1602-1608 (1989), the entirety of which is
herein incorporated by reference; Dotson et al., Plant Physiology
93:98-104 (1989), the entirety of which is herein incorporated by
reference). Homoserine kinase isolated from barley and wheat has
not been reported to exhibit aspartate-derived amino acid feedback
inhibition (Gengenbach et al., Crop Science 18:472-476 (1978);
Dotson et al, Plant Physiology 93:98-104 (1989)). It has been
reported that homoserine kinase exhibits feedback regulation in the
dicots, pea (Rees et al., Biochemical Journal 309:999-1007 (1995),
the entirety of which is herein incorporated by reference) and
radish (Bryan et al., Biochemistry and Biophysics Research
Communications 41:1211-1217 (1970)). Bacterial and yeast homologues
have been reported (Azevedo et al., Phytochemistry 31:3725-3730
(1992); Azevedo et al., Phytochemistry 31:3731-3734 (1992);
Brennecke et al., Phytochemistry 41:707 (1996); Aarnes, Plant
Science Letters 9:137-145 (1977)).
[0010] Sulfur, in yeast, is incorporated into O-acetylhomoserine
resulting in homocysteine. This reaction is catalyzed by the
O-acetylhomoserine sulfhydrylase (EC 4.2.99.10) (also known as
O-acethomoserine (thiol)-lyase). O-acetylhomoserine sulfhydrylase
has been reported to be a homotetramer with a molecular weight of
200,000. O-acetylhomoserine sulfhydrylase has also been reported to
bind four molecules of pyridoxal phosphate (Thomas and
Surdin-Kerjan, Microbiol. Mol. Biol. Rev. 61:503-532 (1997)).
[0011] In higher plants, the sulfur atom from cysteine and the
carbon backbone derived from aspartate used to synthesize
methionine are reported to be catalyzed by pyridoxal 5'-phophate
(PLP) dependent enzymes (Ravanel et al., Proc. Natl. Acad. Sci.
(U.S.A.) 95:7805-7812 (1998), the entirety of which is herein
incorporated by reference). The amino acid composition of the
O-acetylhomoserine sulfhydrylase has also been reported to share
sequence similarities to the E. coli cystathionine .gamma.-synthase
and cystathionine .beta.-lyase and cystathionine .gamma.-lyase from
Saccharomyces cervisiae and rats. All of these enzymes thus appear
to belong to one protein family, whose members have evolved from an
ancestral pyridoxal phosphate enzyme (Thomas and Surdin-Kerjan,
Microbiol. Mol. Biol. Rev. 61:503-532 (1997)).
[0012] In yeast, the synthesis of cysteine from homocysteine has
been reported to require two successive steps, .beta. addition and
.gamma. elimination. Cystathionine .beta.-synthase (EC 4.2.1.22)
has been reported to catalyze the first reaction where homocysteine
and serine yield cystathionine. In S. cervisiae, cystathionine
.beta.-synthase is encoded by STR4. STR4 encodes a polypeptide of
506 residues which shows extensive sequence similarity to its
functional analog in rats. The rat analog has been reported to
contain an additional amino-terminal extension of 60 residues.
Moreover, the two enzymes have been reported to be closely related
to the cyteine synthase from enteric bacteria and plants (Thomas
and Surdin-Kerjan, Microbiol. Mol. Biol. Rev. 61:503-532
(1997)).
[0013] Cystathionine .gamma.-lyase (EC 4.4.1.1) catalyzes the
.gamma. cleavage of cystationine in yeast, the second reported step
of the biosynthesis of cysteine from homocysteine. Cystathionine
.gamma.-lyase has been reported to have a molecular weight of about
194,000 kd. In S. cerviseae, cystathionine .gamma.-lyase is encoded
by STR1. A mutation in the S. cerviseae cystathionine .gamma.-lyase
gene leads to a nutritional requirement for cysteine or
glutathione. The yeast cystathionine .gamma.-lyase belongs to a
protein family which includes a functional analog in rats, a Met25p
from yeast and cystathionin .beta.-lyase and cystathionin
.gamma.-synthase from E. coli (Thomas and Surdin-Kerjan, Microbiol.
Mol. Biol. Rev. 61:503-532 (1997)).
[0014] Cystathionine .gamma.-synthase (also known as
O-succinylhomoserine (thio)-lyase, E.C. 4.2.99.9) catalyzes the
first reported reaction which is unique to methionine biosynthesis,
thereby committing aspartate pathway flux toward this amino acid.
In this reaction, 0-phosphohomoserine and cysteine serve as
substrates for the production of cystathionine. Cystathionine
.gamma.-synthase has not been reported to be regulated by
aspartate-derived amino acids feedback inhibition (Bright et al.,
Biochemical Genetics 20:229-243 (1982); Arruda et al., Plant
Physiology 76:442-446 (1984)). Cystathionine .gamma.-synthase has
however, been reported to be sensitive to product inhibition by
orthophosphate (Lea et al., Barley: Genetics, Molecular Biology and
Biotechnology, Shewrey (ed.), CAB International, Oxford, 181
(1992); Davies et al., Plant Science Letters 9:323-332 (1977)).
Cloned cystathionine .gamma.-synthase have been reported from
Arabidopsis thaliana (Davies et al., Plant Physiology 62:536-541
(1978)). It has been reported that methionine levels are modulated
via regulation of cystathionine-synthase (Matthews et al.,
Zeitschrift fur Naturforschung, Section Bioscience 34:1177-1185
(1979-2724 (1974); Lea et al., FEBS Letters 98:165 (1979), all of
which references are incorporated herein in their entirety).
[0015] Cystathionine .beta.-lyase catalyzes the next reaction in
the biosynthesis of methionine. This reaction generates
homocysteine, pyruvate and ammonia from the enzymatic decomposition
of cystathionine. Evidence for isoenzymes which differ with respect
to cellular localization have been reported for barley (Matthews et
al., Canadian Journal of Botany 57:299-304 (1979)) and spinach
(Rognes et al., Nature 287:357-359 (1980), the entirety of which is
herein incorporated by reference).
[0016] De novo synthesis of methionine from homocysteine uses a
methyl group which originates from single-carbon metabolism. In
this metabolism, derivatives of tetrahydrofolate transfer
one-carbon groups at the oxidation levels of methanol, fomaldehyde
and formate to acceptor molecules. Single-carbon derivatives of
tetrahydrofolate are required for the biosynthesis of methionine,
purine nucleotides and thymidylate as well as for the synthesis of
N-formylmethionine in the mitochondrion. S. cerevisiae possesses
two complete sets of folate interconversion enzymes, one located in
the cytosol (methionyl-tRNA synthetase, EC 6.1.1.10) and the other
located in the mitochondrion (methionyl t-RNA synthetase, EC
6.1.1.10) (Thomas and Surdin-Kerjan, Microbiol. Mol. Biol. Rev.
61:503-532 (1997)) and in plants including the chloroplast (Menand
et al., Proc. Natl. Acad. Sci. (U.S.A.), 95:11014-11019 (1998), the
entirety of which is herein incorporated by reference).
[0017] Methionine synthase generates methionine from homocysteine
by a methylation reaction and thus represents the final step of the
methionine biosynthetic pathway. Methionine synthase is also
sometimes referred to as
5-methyltetrahydropteroyltriglutamate-homocysteine-S-methyltransferase-
. N-methyltetrahydrofolate serves as the methyl donor in this
reaction, which occurs in the absence of cobalamin (Giovanelli et
al., Plant Physiology 90:1577-1583 (1989), the entirety of which is
herein incorporated by reference; Green et al., Crop Science
14:827-830 (1974), the entirety of which is herein incorporated by
reference).
II. Methoinine Degradation Pathway
[0018] Plants contain a pathway for the degradation of
L-methionine. This degradation pathway includes the following
enzymes: methionine adenosyltransferase (EC 2.5.1.6), methionine
S-methyltransferase (EC 2.1.1.12), adenosylmethionine hydrolase (EC
3.3.1.2), homocysteine S-methyltransferase (EC 2.1.1.10) and
S-adenosyl-methionine decarboxylase (EC 4.1.1.50).
[0019] The reported first step in the catabolism of methionine is
the ATP-dependent conversion to S-adenosylmethionine (AdoMet),
which is catalyzed by the enzyme methionine adenosyltransferase,
also known as S-adenosylmethionine synthetase. Methionine
adenosyltransferase enzyme has been characterized from several
plant sources (Aames, Plant Science Letters 10:381 (1977), the
entirety of which is herein incorporated by reference; Mathur et
al., Biochimia and Biophysica Acta 1078:161-170 (1991), the
entirety of which is herein incorporated by reference; Kim et al.,
Journal of Biochemical and Molecular Biology 28:100 (1995), the
entirety of which is herein incorporated by reference) and nucleic
acid molecules (genomic and cDNA) have also been obtained from a
variety of sources (Izhaki et al., Plant Physiology 108:841-842
(1995), the entirety of which is herein incorporated by reference;
Espartero et al., Molecular Biology Plant 25:217-237 (1994), the
entirety of which is herein incorporated by reference). Regulation
of methionine adenosyltransferase activity has been observed for
the enzyme from Glycine max (soybean). In Glycine max, methionine
adenosyltransferase was reportedly inhibited by
S-adenosylmethionine (Kim et al., Journal of Biochemical and
Molecular Biology 28:100 (1995). Studies have also reported that
the levels of methionine adenosyltransferase appear to fluctuate in
response to hormonal or environmental conditions such as
gibberellic acid (Mathur et al., Biochimica and Biophysica
Acta/162:289-290 (1993), the entirety of which is herein
incorporated by reference; Mathur et al., Biochimica and Biophysica
Acta 1137:338-348 (1992), the entirety of which is herein
incorporated by reference), salt stress (Espartero et al.,
Molecular Biology Plant 25:217-227 (1994) the entirety of which is
herein incorporated by reference) and wounding (Kim et al., Plant
Cell Reports 13:340 (1994), the entirety of which is herein
incorporated by reference). It has also been reported that
methionine adenosyltransferase may play a role in the lignification
process (Peleman et al., Plant Cell 1:81 (1989), the entirety of
which is herein incorporated by reference).
[0020] AdoMet is further catabolized by several enzymes and has
been reported to serve a variety of metabolic functions including
that of a methyl donor (Cossins, The Biochemistry of Plants 11:317
Devis (ed.), Academic Press, San Diego (1987), the entirety of
which is herein incorporated by reference) that of a precursor for
polyamine biosynthesis (Tiburico et al., The Biochemistry of Plants
16:283 (1990), the entirety of which is herein incorporated by
reference) and that of a precursor for ethylene biosynthesis
(Kende, Plant Physiology 91:1-4 (1989), the entirety of which is
herein incorporated by reference; Flurh et al., Critical Review of
Plant Science 15:479 (1996), the entirety of which is herein
incorporated by reference). In each case, enzymes are present to
regenerate methionine from the sulfur-containing backbone resulting
in no net loss of methionine.
[0021] An enzyme involved in AdoMet catabolism is
adenosylmethionine hydrolase (EC 3.3.1.2) which converts AdoMet to
methylthioadenosine and L-homoserine. L-homoserine is further
metabolized during the biosynthesis of polyamines and ethylene and
methylthioadenosine is recycled to methionine. In yeast, a form of
adenosylmethionine hydrolase (EC 3.1.1.1) has been reported
(http://www.ncbi.nlm.nih.gov/htbin-post/Entrez/query (1998)).
[0022] Another enzyme for which AdoMet is a substrate for is
homocysteine S-methyltransferase. Homocysteine S-methyltransferase
catalyzes the combination of AdoMet, with L-homocysteine to produce
both S-adenosyl-L-homocysteine and L-methionine. Another enzyme has
been described which generates S-adenosyl-L-homocysteine from
AdoMet. This enzyme is called methionine S-methyltransferase and it
catalyzes the reaction in which S-adenosyl-L-homocysteine reacts
with L-methionine to generate S-adenosyl-L-homocysteine and
S-methyl-L-methionine. AdoMet can also be decarboxylated by
adenosyl methionine decarboxylase, which generates
(5-deoxy-5-adenosyl) (3-aminopropyl) methylsulfonium salt.
III. Expressed Sequence Tag Nucleic Acid Molecules
[0023] Expressed sequence tags, or ESTs are randomly sequenced
members of a cDNA library (or complementary DNA)(McCombie et al.,
Nature Genetics 1:124-130 (1992); Kurata et al., Nature Genetics
8:365-372 (1994); Okubo et al., Nature Genetics 2:173-179 (1992),
all of which references are incorporated herein in their entirety).
The randomly selected clones comprise insets that can represent a
copy of up to the full length of a mRNA transcript.
[0024] Using conventional methodologies, cDNA libraries can be
constructed from the mRNA (messenger RNA) of a given tissue or
organism using poly dT primers and reverse transcriptase
(Efstratiadis et al., Cell 7:279-3680 (1976), the entirety of which
is herein incorporated by reference; Higuchi et al., Proc. Natl.
Acad. Sci. (U.S.A.) 73:3146-3150 (1976), the entirety of which is
herein incorporated by reference; Maniatis et al., Cell 8:163-182
(1976) the entirety of which is herein incorporated by reference;
Land et al., Nucleic Acids Res. 9:2251-2266 (1981), the entirety of
which is herein incorporated by reference; Okayama et al., Mol.
Cell. Biol. 2:161-170 (1982), the entirety of which is herein
incorporated by reference; Gubler et al., Gene 25:263-269 (1983),
the entirety of which is herein incorporated by reference).
[0025] Several methods may be employed to obtain full-length cDNA
constructs. For example, terminal transferase can be used to add
homopolymeric tails of dC residues to the free 3' hydroxyl groups
(Land et al., Nucleic Acids Res. 9:2251-2266 (1981), the entirety
of which is herein incorporated by reference). This tail can then
be hybridized by a poly dG oligo which can act as a primer for the
synthesis of full length second strand cDNA. Okayama and Berg, Mol.
Cell. Biol. 2:161-170 (1982), the entirety of which is herein
incorporated by reference, report a method for obtaining full
length cDNA constructs. This method has been simplified by using
synthetic primer-adapters that have both homopolymeric tails for
priming the synthesis of the first and second strands and
restriction sites for cloning into plasmids (Coleclough et al.,
Gene 34:305-314 (1985), the entirety of which is herein
incorporated by reference) and bacteriophage vectors (Krawinkel et
al., Nucleic Acids Res. 14:1913 (1986), the entirety of which is
herein incorporated by reference; Han et al., Nucleic Acids Res.
15:6304 (1987), the entirety of which is herein incorporated by
reference).
[0026] These strategies have been coupled with additional
strategies for isolating rare mRNA populations. For example, a
typical mammalian cell contains between 10,000 and 30,000 different
mRNA sequences (Davidson, Gene Activity in Early Development, 2nd
ed., Academic Press, New York (1976), the entirety of which is
herein incorporated by reference). The number of clones required to
achieve a given probability that a low-abundance mRNA will be
present in a cDNA library is N=(ln(1-P))/(ln(1-1/n)) where N is the
number of clones required, P is the probability desired and 1/n is
the fractional proportion of the total mRNA that is represented by
a single rare mRNA (Sambrook et al., Molecular Cloning: A
Laboratory Manual, 2nd ed., Cold Spring Harbor Laboratory Press
(1989), the entirety of which is herein incorporated by
reference).
[0027] A method to enrich preparations of mRNA for sequences of
interest is to fractionate by size. One such method is to
fractionate by electrophoresis through an agarose gel (Pennica et
al., Nature 301:214-221 (1983), the entirety of which is herein
incorporated by reference). Another such method employs sucrose
gradient centrifugation in the presence of an agent, such as
methylmercuric hydroxide, that denatures secondary structure in RNA
(Schweinfest et al., Proc. Natl. Acad. Sci. (U.S.A.) 79:4997-5000
(1982), the entirety of which is herein incorporated by
reference).
[0028] A frequently adopted method is to construct equalized or
normalized cDNA libraries (Ko, Nucleic Acids Res. 18:5705-5711
(1990), the entirety of which is herein incorporated by reference;
Patanjali et al., Proc. Natl. Acad. Sci. (U.S.A.) 88:1943-1947
(1991), the entirety of which is herein incorporated by reference).
Typically, the cDNA population is normalized by subtractive
hybridization (Schmid et al., J. Neurochem. 48:307-312 (1987), the
entirety of which is herein incorporated by reference; Fargnoli et
al., Anal. Biochem. 187:364-373 (1990), the entirety of which is
herein incorporated by reference; Travis et al., Proc. Natl. Acad.
Sci. (U.S.A.) 85:1696-1700 (1988), the entirety of which is herein
incorporated by reference; Kato, Eur. J. Neurosci. 2:704-711
(1990); and Schweinfest et al., Genet. Anal. Tech. Appl. 7:64-70
(1990), the entirety of which is herein incorporated by reference).
Subtraction represents another method for reducing the population
of certain sequences in the cDNA library (Swaroop et al., Nucleic
Acids Res. 19:1954 (1991), the entirety of which is herein
incorporated by reference).
[0029] ESTs can be sequenced by a number of methods. Two basic
methods may be used for DNA sequencing, the chain termination
method of Sanger et al., Proc. Natl. Acad. Sci. (U.S.A.)
74:5463-5467 (1977), the entirety of which is herein incorporated
by reference and the chemical degradation method of Maxam and
Gilbert, Proc. Nat. Acad. Sci. (U.S.A.) 74:560-564 (1977), the
entirety of which is herein incorporated by reference. Automation
and advances in technology such as the replacement of radioisotopes
with fluorescence-based sequencing have reduced the effort required
to sequence DNA (Craxton, Methods 2:20-26 (1991), the entirety of
which is herein incorporated by reference; Ju et al., Proc. Natl.
Acad. Sci. (U.S.A.) 92:4347-4351 (1995), the entirety of which is
herein incorporated by reference; Tabor and Richardson, Proc. Natl.
Acad. Sci. (U.S.A.) 92:6339-6343 (1995), the entirety of which is
herein incorporated by reference). Automated sequencers are
available from, for example, Pharmacia Biotech, Inc., Piscataway,
N.J. (Pharmacia ALF), LI-COR, Inc., Lincoln, Nebr. (LI-COR 4,000)
and Millipore, Bedford, Mass. (Millipore BaseStation).
[0030] In addition, advances in capillary gel electrophoresis have
also reduced the effort required to sequence DNA and such advances
provide a rapid high resolution approach for sequencing DNA samples
(Swerdlow and Gesteland, Nucleic Acids Res. 18:1415-1419 (1990);
Smith, Nature 349:812-813 (1991); Luckey et al., Methods Enzymol.
218:154-172 (1993); Lu et al., J. Chromatog. A. 680:497-501 (1994);
Carson et al., Anal. Chem. 65:3219-3226 (1993); Huang et al., Anal.
Chem. 64:2149-2154 (1992); Kheterpal et al., Electrophoresis
17:1852-1859 (1996); Quesada and Zhang, Electrophoresis
17:1841-1851 (1996); Baba, Yakugaku Zasshi 117:265-281 (1997), all
of which are herein incorporated by reference in their
entirety).
[0031] ESTs longer than 150 nucleotides have been found to be
useful for similarity searches and mapping (Adams et al., Science
252:1651-1656 (1991), herein incorporated by reference). ESTs,
which can represent copies of up to the full length transcript, may
be partially or completely sequenced. Between 150-450 nucleotides
of sequence information is usually generated as this is the length
of sequence information that is routinely and reliably produced
using single run sequence data. Typically, only single run sequence
data is obtained from the cDNA library (Adams et al., Science
252:1651-1656 (1991). Automated single run sequencing typically
results in an approximately 2-3% error or base ambiguity rate
(Boguski et al., Nature Genetics 4:332-333 (1993), the entirety of
which is herein incorporated by reference).
[0032] EST databases have been constructed or partially constructed
from, for example, C. elegans (McCombrie et al., Nature Genetics
1:124-131 (1992)), human liver cell line HepG2 (Okubo et al.,
Nature Genetics 2:173-179 (1992)), human brain RNA (Adams et al.,
Science 252:1651-1656 (1991); Adams et al., Nature 355:632-635
(1992)), Arabidopsis, (Newman et al., Plant Physiol. 106:1241-1255
(1994)); and rice (Kurata et al., Nature Genetics 8:365-372
(1994)).
IV. Sequence Comparisons
[0033] A characteristic feature of a DNA sequence is that it can be
compared with other DNA sequences. Sequence comparisons can be
undertaken by determining the similarity of the test or query
sequence with sequences in publicly available or proprietary
databases ("similarity analysis") or by searching for certain
motifs ("intrinsic sequence analysis")(e.g. cis elements)(Coulson,
Trends in Biotechnology 12:76-80 (1994), the entirety of which is
herein incorporated by reference); Birren et al., Genome Analysis
1: Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y.
543-559 (1997), the entirety of which is herein incorporated by
reference).
[0034] Similarity analysis includes database search and alignment.
Examples of public databases include the DNA Database of Japan
(DDBJ)(http://www.ddbj.nig.ac.jp/); Genebank
(http://www.ncbi.nlm.nih.gov/Web/Search/Index.htlm); and the
European Molecular Biology Laboratory Nucleic Acid Sequence
Database (EMBL)
(http://www.ebi.ac.uk/ebi_docs/embl_db/embl-db.html). Other
appropriate databases include dbEST
(http://www.ncbi.nlm.nih.gov/dbEST/index.html), SwissProt
(http://www.ebi.ac.uk/ebi_docs/swisprot_db/swisshome.html), PIR
(http://www-nbrt.georgetown.edu/pir/) and The Institute for Genome
Research (http://www.tigr.org/tdb/tdb.html)
[0035] A number of different search algorithms have been developed,
one example of which are the suite of programs referred to as BLAST
programs. There are five implementations of BLAST, three designed
for nucleotide sequences queries (BLASTN, BLASTX and TBLASTX) and
two designed for protein sequence queries (BLASTP and TBLASTN)
(Coulson, Trends in Biotechnology 12:76-80 (1994); Birren et al.,
Genome Analysis 1, Cold Spring Harbor Laboratory Press, Cold Spring
Harbor, N.Y. 543-559 (1997)).
[0036] BLASTN takes a nucleotide sequence (the query sequence) and
its reverse complement and searches them against a nucleotide
sequence database. BLASTN was designed for speed, not maximum
sensitivity and may not find distantly related coding sequences.
BLASTX takes a nucleotide sequence, translates it in three forward
reading frames and three reverse complement reading frames and then
compares the six translations against a protein sequence database.
BLASTX is useful for sensitive analysis of preliminary
(single-pass) sequence data and is tolerant of sequencing errors
(Gish and States, Nature Genetics 3:266-272 (1993), the entirety of
which is herein incorporated by reference). BLASTN and BLASTX may
be used in concert for analyzing EST data (Coulson, Trends in
Biotechnology 12:76-80 (1994); Birren et al., Genome Analysis
1:543-559 (1997)).
[0037] Given a coding nucleotide sequence and the protein it
encodes, it is often preferable to use the protein as the query
sequence to search a database because of the greatly increased
sensitivity to detect more subtle relationships. This is due to the
larger alphabet of proteins (20 amino acids) compared with the
alphabet of nucleic acid sequences (4 bases), where it is far
easier to obtain a match by chance. In addition, with nucleotide
alignments, only a match (positive score) or a mismatch (negative
score) is obtained, but with proteins, the presence of conservative
amino acid substitutions can be taken into account. Here, a
mismatch may yield a positive score if the non-identical residue
has physical/chemical properties similar to the one it replaced.
Various scoring matrices are used to supply the substitution scores
of all possible amino acid pairs. A general purpose scoring system
is the BLOSUM62 matrix (Henikoff and Henikoff, Proteins 17:49-61
(1993), the entirety of which is herein incorporated by reference),
which is currently the default choice for BLAST programs. BLOSUM62
is tailored for alignments of moderately diverged sequences and
thus may not yield the best results under all conditions. Altschul,
J. Mol. Biol. 36:290-300 (1993), the entirety of which is herein
incorporated by reference, describes a combination of three
matrices to cover all contingencies. This may improve sensitivity,
but at the expense of slower searches. In practice, a single
BLOSUM62 matrix is often used but others (PAM40 and PAM250) may be
attempted when additional analysis is necessary. Low PAM matrices
are directed at detecting very strong but localized sequence
similarities, whereas high PAM matrices are directed at detecting
long but weak alignments between very distantly related
sequences.
[0038] Homologues in other organisms are available that can be used
for comparative sequence analysis. Multiple alignments are
performed to study similarities and differences in a group of
related sequences. CLUSTAL W is a multiple sequence alignment
package that performs progressive multiple sequence alignments
based on the method of Feng and Doolittle, J. Mol. Evol. 25:351-360
(1987), the entirety of which is herein incorporated by reference.
Each pair of sequences is aligned and the distance between each
pair is calculated; from this distance matrix, a guide tree is
calculated and all of the sequences are progressively aligned based
on this tree. A feature of the program is its sensitivity to the
effect of gaps on the alignment; gap penalties are varied to
encourage the insertion of gaps in probable loop regions instead of
in the middle of structured regions. Users can specify gap
penalties, choose between a number of scoring matrices, or supply
their own scoring matrix for both pairwise alignments and multiple
alignments. CLUSTAL W for UNIX and VMS systems is available at:
ftp.ebi.ac.uk. Another program is MACAW (Schuler et al., Proteins
Struct. Func. Genet. 9:180-190 (1991), the entirety of which is
herein incorporated by reference, for which both Macintosh and
Microsoft Windows versions are available. MACAW uses a graphical
interface, provides a choice of several alignment algorithms and is
available by anonymous ftp at: ncbi.nlm.nih.gov
(directory/pub/macaw).
[0039] Sequence motifs are derived from multiple alignments and can
be used to examine individual sequences or an entire database for
subtle patterns. With motifs, it is sometimes possible to detect
distant relationships that may not be demonstrable based on
comparisons of primary sequences alone. Currently, the largest
collection of sequence motifs in the world is PROSITE (Bairoch and
Bucher, Nucleic Acid Research 22:3583-3589 (1994), the entirety of
which is herein incorporated by reference). PROSITE may be accessed
via either the ExPASy server on the World Wide Web or anonymous ftp
site. Many commercial sequence analysis packages also provide
search programs that use PROSITE data.
[0040] A resource for searching protein motifs is the BLOCKS E-mail
server developed by Henikoff, Trends Biochem Sci. 18:267-268
(1993), the entirety of which is herein incorporated by reference;
Henikoff and Henikoff, Nucleic Acid Research 19:6565-6572 (1991),
the entirety of which is herein incorporated by reference; Henikoff
and Henikoff, Proteins 17:49-61 (1993). BLOCKS searches a protein
or nucleotide sequence against a database of protein motifs or
"blocks." Blocks are defined as short, =gapped multiple alignments
that represent highly conserved protein patterns. The blocks
themselves are derived from entries in PROSITE as well as other
sources. Either a protein query or a nucleotide query can be
submitted to the BLOCKS server; if a nucleotide sequence is
submitted, the sequence is translated in all six reading frames and
motifs are sought for these conceptual translations. Once the
search is completed, the server will return a ranked list of
significant matches, along with an alignment of the query sequence
to the matched BLOCKS entries.
[0041] Conserved protein domains can be represented by
two-dimensional matrices, which measure either the frequency or
probability of the occurrences of each amino acid residue and
deletions or insertions in each position of the domain. This type
of model, when used to search against protein databases, is
sensitive and usually yields more accurate results than simple
motif searches. Two popular implementations of this approach are
profile searches such as GCG program ProfileSearch and Hidden
Markov Models (HMMs)(Krough et al., J. Mol. Biol. 235:1501-1531,
(1994); Eddy, Current Opinion in Structural Biology 6:361-365,
(1996), both of which are herein incorporated by reference in their
entirety). In both cases, a large number of common protein domains
have been converted into profiles, as present in the PROSITE
library, or HHM models, as in the Pfam protein domain library
(Sonnhammer et al., Proteins 28:405-420 (1997), the entirety of
which is herein incorporated by reference). Pfam contains more than
500 HMM models for enzymes, transcription factors, signal
transduction molecules and structural proteins. Protein databases
can be queried with these profiles or HMM models, which will
identify proteins containing the domain of interest. For example,
HMMSW or HMMFS, two programs in a public domain package called
HMMER (Sonnhammer et al., Proteins 28:405-420 (1997)) can be
used.
[0042] PROSITE and BLOCKS represent collected families of protein
motifs. Thus, searching these databases entails submitting a single
sequence to determine whether or not that sequence is similar to
the members of an established family. Programs working in the
opposite direction compare a collection of sequences with
individual entries in the protein databases. An example of such a
program is the Motif Search Tool, or MoST (Tatusov et al., Proc.
Natl. Acad. Sci. (U.S.A.) 91:12091-12095 (1994), the entirety of
which is herein incorporated by reference). On the basis of an
aligned set of input sequences, a weight matrix is calculated by
using one of four methods (selected by the user). A weight matrix
is simply a representation, position by position of how likely a
particular amino acid will appear. The calculated weight matrix is
then used to search the databases. To increase sensitivity, newly
found sequences are added to the original data set, the weight
matrix is recalculated and the search is performed again. This
procedure continues until no new sequences are found.
SUMMARY OF THE INVENTION
[0043] The present invention provides a substantially purified
nucleic acid molecule that encodes a maize or soybean enzyme or
fragment thereof, wherein said maize or soybean enzyme is selected
from the group consisting of: (a) methionine adenosyltransferase,
(b) S-adenosyl-methionine decarboxylase, (c) aspartate kinase, (d)
aspartate-semialdehyde dehydrogenase, (e) cystathionine
gamma-synthase, (f) cystathionine beta-lysase, and (g)
5-methyltetrahydropteroyltriglutamate-homocysteine-S-methyltransferase.
[0044] The present invention also provides a substantially purified
nucleic acid molecule that encodes a plant methionine pathway
enzyme or fragment thereof, wherein the nucleic acid molecule is
selected from the group consisting of a nucleic acid molecule that
encodes a maize or a soybean methionine adenosyltransferase enzyme
or fragment thereof, a nucleic acid molecule that encodes a maize
or a soybean S-adenosylmethionine decarboxylase enzyme or fragment
thereof, a nucleic acid molecule that encodes a maize or a soybean
aspartate kinase enzyme or fragment thereof, a nucleic acid
molecule that encodes a maize or a soybean aspartate-semialdehyde
dehydrogenase enzyme or fragment thereof, a nucleic acid molecule
that encodes a maize or a soybean O-succinylhomoserine
(thiol)-lyase enzyme or fragment thereof, a nucleic acid molecule
that encodes a maize or a soybean cystathionine .beta.-lyase enzyme
or fragment thereof, a nucleic acid molecule that encodes a maize
or a soybean
5-methyltetrahydropteroyltriglutamate-homocysteine-S-methyltransferase
enzyme or fragment thereof, a nucleic acid molecule that encodes a
maize or a soybean adenosylhomocysteinase enzyme or fragment
thereof, a nucleic acid molecule that encodes a maize or a soybean
cystathionine .beta.-synthase enzyme or fragment thereof, a nucleic
acid molecule that encodes a maize or a soybean cystathionine
.gamma.-lyase enzyme or fragment thereof and a nucleic acid
molecule that encodes a maize or a soybean O-acetylhomoserine
(thiol)-lyase enzyme or fragment thereof.
[0045] A substantially purified maize or soybean enzyme or fragment
thereof, wherein said maize or soybean enzyme is selected from the
group consisting of (a) methionine adenosyltransferase or fragment
thereof; (b) S-adenosyl-methionine decarboxylase or fragment
thereof; (c) aspartate kinase or fragment thereof; (d)
aspartate-semialdehyde dehydrogenase or fragment thereof; (e)
cystathionine gamma-synthase or fragment thereof; (f) cystathionine
beta-lysase or fragment thereof; and (e)
5-methyltetrahydropteroyltriglutamate-homocysteine-S-methyltransferase
or fragment thereof.
[0046] The present invention also provides a substantially purified
maize or soybean methionine pathway protein or fragment thereof
encoded by a first nucleic acid molecule which specifically
hybridizes to a second nucleic acid molecule, the second nucleic
acid molecule having a nucleic acid sequence selected from the
group consisting of a complement of SEQ ID NO: 1 through SEQ ID NO:
3204.
[0047] The present invention also provides a substantially purified
maize or soybean methionine adenosyltransferase enzyme or fragment
thereof encoded by a first nucleic acid molecule which specifically
hybridizes to a second nucleic acid molecule, the second nucleic
acid molecule having a nucleic acid sequence selected from the
group consisting of a complement of SEQ ID NO: 1 through SEQ ID NO:
429 and SEQ ID NO: 1635 through SEQ ID NO: 2479.
[0048] The present invention also provides a substantially purified
maize or soybean methionine adenosyltransferase enzyme or fragment
thereof encoded by a nucleic acid sequence selected from the group
consisting of SEQ ID NO: 1 through SEQ ID NO: 429 and SEQ ID NO:
1635 through SEQ ID NO: 2479.
[0049] The present invention also provides a substantially purified
maize or soybean S-adenosylmethionine decarboxylase enzyme or
fragment thereof encoded by a first nucleic acid molecule which
specifically hybridizes to a second nucleic acid molecule, the
second nucleic acid molecule having a nucleic acid sequence
selected from the group consisting of a complement of SEQ ID NO:
430 through SEQ ID NO: 857 and SEQ ID NO: 2480 through SEQ ID NO:
2623.
[0050] The present invention also provides a substantially purified
maize or soybean S-adenosylmethionine decarboxylase enzyme or
fragment thereof encoded by a nucleic acid sequence selected from
the group consisting of SEQ ID NO: 430 through SEQ ID NO: 857 and
SEQ ID NO: 2480 through SEQ ID NO: 2623.
[0051] The present invention also provides a substantially purified
maize or soybean aspartate kinase enzyme or fragment thereof
encoded by a first nucleic acid molecule which specifically
hybridizes to a second nucleic acid molecule, the second nucleic
acid molecule having a nucleic acid sequence selected from the
group consisting of a complement of SEQ ID NO: 858 through SEQ ID
NO: 900 and SEQ ID NO: 2624 through SEQ ID NO: 2648.
[0052] The present invention also provides a substantially purified
maize or soybean aspartate kinase enzyme or fragment thereof
encoded by a nucleic acid sequence selected from the group
consisting of SEQ ID NO: 858 through SEQ ID NO: 900 and SEQ ID NO:
2624 through SEQ ID NO: 2648.
[0053] The present invention also provides a substantially purified
maize or soybean aspartate-semialdehyde dehydrogenase enzyme or
fragment thereof encoded by a first nucleic acid molecule which
specifically hybridizes to a second nucleic acid molecule, the
second nucleic acid molecule having a nucleic acid sequence
selected from the group consisting of a complement of SEQ ID NO:
901 through SEQ ID NO: 904 and SEQ ID NO: 2649 through SEQ ID NO:
2654.
[0054] The present invention also provides a substantially purified
maize or soybean aspartate-semialdehyde dehydrogenase enzyme or
fragment thereof encoded by a nucleic acid sequence selected from
the group consisting of SEQ ID NO: 901 through SEQ ID NO: 904 and
SEQ ID NO: 2649 through SEQ ID NO: 2654.
[0055] The present invention also provides a substantially purified
maize or soybean O-succinylhomoserine (thiol)-lyase enzyme or
fragment thereof encoded by a first nucleic acid molecule which
specifically hybridizes to a second nucleic acid molecule, the
second nucleic acid molecule having a nucleic acid sequence
selected from the group consisting of a complement of SEQ ID NO:
905 through SEQ ID NO: 953 and SEQ ID NO: 2655 through SEQ ID NO:
2660.
[0056] The present invention also provides a substantially purified
maize or soybean O-succinylhomoserine (thiol)-lyase enzyme or
fragment thereof encoded by a nucleic acid sequence selected from
the group consisting of SEQ ID NO: 905 through SEQ ID NO: 953 and
SEQ ID NO: 2655 through SEQ ID NO: 2660.
[0057] The present invention also provides a substantially purified
maize or soybean cystathionine .beta.-lyase enzyme or fragment
thereof encoded by a first nucleic acid molecule which specifically
hybridizes to a second nucleic acid molecule, the second nucleic
acid molecule having a nucleic acid sequence selected from the
group consisting of a complement of SEQ ID NO: 954 through SEQ ID
NO: 963 and SEQ ID NO: 2661 through SEQ ID NO: 2665.
[0058] The present invention also provides a substantially purified
maize or soybean cystathionine .beta.-lyase enzyme or fragment
thereof encoded by a nucleic acid sequence selected from the group
consisting of SEQ ID NO: 954 through SEQ ID NO: 963 and SEQ ID NO:
2661 through SEQ ID NO: 2665.
[0059] The present invention also provides a substantially purified
maize or soybean
5-methyltetrahydropteroyltriglutamate-homocysteine-S-methyltransferase
enzyme or fragment thereof encoded by a first nucleic acid molecule
which specifically hybridizes to a second nucleic acid molecule,
the second nucleic acid molecule having a nucleic acid sequence
selected from the group consisting of a complement of SEQ ID NO:
964 through SEQ ID NO: 1353 and SEQ ID NO: 2666 through SEQ ID NO:
2992.
[0060] The present invention also provides a substantially purified
maize or soybean
5-methyltetrahydropteroyltriglutamate-homocysteine-S-methyltransferase
enzyme or fragment thereof encoded by a nucleic acid sequence
selected from the group consisting of SEQ ID NO: 964 through SEQ ID
NO: 1353 and SEQ ID NO: 2666 through SEQ ID NO: 2992.
[0061] The present invention also provides a substantially purified
maize or adenosylhomocysteinase enzyme or fragment thereof encoded
by a first nucleic acid molecule which specifically hybridizes to a
second nucleic acid molecule, the second nucleic acid molecule
having a nucleic acid sequence selected from the group consisting
of a complement of SEQ ID NO: 1354 through SEQ ID NO: 1630 and SEQ
ID NO: 2993 through SEQ ID NO: 3199.
[0062] The present invention also provides a substantially purified
maize or adenosylhomocysteinase enzyme or fragment thereof encoded
by a nucleic acid sequence selected from the group consisting of
SEQ ID NO: 1354 through SEQ ID NO: 1630 and SEQ ID NO: 2993 through
SEQ ID NO: 3199.
[0063] The present invention also provides a substantially purified
maize or cystathionine (3-synthase enzyme or fragment thereof
encoded by a first nucleic acid molecule which specifically
hybridizes to a second nucleic acid molecule, the second nucleic
acid molecule having a nucleic acid sequence selected from the
group consisting of a complement of SEQ ID NO: 1631 through SEQ ID
NO: 1632.
[0064] The present invention also provides a substantially purified
maize or cystathionine (3-synthase enzyme or fragment thereof
encoded by a nucleic acid sequence selected from the group
consisting of SEQ ID NO: 1631 through SEQ ID NO: 1632.
[0065] The present invention also provides a substantially purified
maize or cystathionine .gamma.-lyase enzyme or fragment thereof
encoded by a first nucleic acid molecule which specifically
hybridizes to a second nucleic acid molecule, the second nucleic
acid molecule having a nucleic acid sequence selected from the
group consisting of a complement of SEQ ID NO: 1633 through SEQ ID
NO: 1634 and SEQ ID NO: 3203 through SEQ ID NO: 3204.
[0066] The present invention also provides a substantially purified
maize or cystathionine .gamma.-lyase enzyme or fragment thereof
encoded by a nucleic acid sequence selected from the group
consisting of SEQ ID NO: 1633 through SEQ ID NO: 1634 and SEQ ID
NO: 3203 through SEQ ID NO: 3204.
[0067] The present invention also provides a substantially purified
maize or O-acetylhomoserine (thiol)-lyase enzyme or fragment
thereof encoded by a first nucleic acid molecule which specifically
hybridizes to a second nucleic acid molecule, the second nucleic
acid molecule having a nucleic acid sequence selected from the
group consisting of a complement of SEQ ID NO: 3200 through SEQ ID
NO: 3202.
[0068] The present invention also provides a substantially purified
maize or O-acetylhomoserine (thiol)-lyase enzyme or fragment
thereof encoded by a nucleic acid sequence selected from the group
consisting of SEQ ID NO: 3200 through SEQ ID NO: 3202.
[0069] The present invention also provides a purified antibody or
fragment thereof which is capable of specifically binding to a
specific maize or soybean enzyme or fragment thereof, wherein said
maize or soybean enzyme or fragment thereof is encoded by a nucleic
acid molecule comprising a nucleic acid sequence selected from the
group consisting of consisting of SEQ ID NO: 1 through SEQ ID NO:
3204.
[0070] The present invention also provides a substantially purified
antibody or fragment thereof, the antibody or fragment thereof
capable of specifically binding to a maize or a soybean methionine
adenosyltransferase enzyme or fragment thereof encoded by a first
nucleic acid molecule which specifically hybridizes to a second
nucleic acid molecule, the second nucleic acid molecule having a
nucleic acid sequence selected from the group consisting of a
complement of SEQ ID NO: 1 through SEQ ID NO: 429 and SEQ ID NO:
1635 through SEQ ID NO: 2479 or a substantially purified maize or
soybean methionine adenosyltransferase enzyme or fragment thereof
encoded by a nucleic acid sequence selected from the group
consisting of SEQ ID NO: 1 through SEQ ID NO: 429 and SEQ ID NO:
1635 through SEQ ID NO: 2479.
[0071] The present invention also provides a substantially purified
antibody or fragment thereof, the antibody or fragment thereof
capable of specifically binding to a maize or a soybean
S-adenosylmethionine decarboxylase enzyme or fragment thereof
encoded by a first nucleic acid molecule which specifically
hybridizes to a second nucleic acid molecule, the second nucleic
acid molecule having a nucleic acid sequence selected from the
group consisting of a complement of SEQ ID NO: 430 through SEQ ID
NO: 857 and SEQ ID NO: 2480 through SEQ ID NO: 2623 or a
substantially purified maize or soybean S-adenosylmethionine
decarboxylase enzyme or fragment thereof encoded by a nucleic acid
sequence selected from the group consisting of SEQ ID NO: 430
through SEQ ID NO: 857 and SEQ ID NO: 2480 through SEQ ID NO:
2623.
[0072] The present invention also provides a substantially purified
antibody or fragment thereof, the antibody or fragment thereof
capable of specifically binding to a maize or a soybean aspartate
kinase enzyme or fragment thereof encoded by a first nucleic acid
molecule which specifically hybridizes to a second nucleic acid
molecule, the second nucleic acid molecule having a nucleic acid
sequence selected from the group consisting of a complement of SEQ
ID NO: 858 through SEQ ID NO: 900 and SEQ ID NO: 2624 through SEQ
ID NO: 2648 or a substantially purified maize or soybean aspartate
kinase enzyme or fragment thereof encoded by a nucleic acid
sequence selected from the group consisting of SEQ ID NO: 858
through SEQ ID NO: 900 and SEQ ID NO: 2624 through SEQ ID NO:
2648.
[0073] The present invention also provides a substantially purified
antibody or fragment thereof, the antibody or fragment thereof
capable of specifically binding to a maize or a soybean
aspartate-semialdehyde dehydrogenase enzyme or fragment thereof
encoded by a first nucleic acid molecule which specifically
hybridizes to a second nucleic acid molecule, the second nucleic
acid molecule having a nucleic acid sequence selected from the
group consisting of a complement of SEQ ID NO: 901 through SEQ ID
NO: 904 and SEQ ID NO: 2649 through SEQ ID NO: 2654 or a
substantially purified maize or soybean enzyme
aspartate-semialdehyde dehydrogenase or fragment thereof encoded by
a nucleic acid sequence selected from the group consisting of SEQ
ID NO: 901 through SEQ ID NO: 904 and SEQ ID NO: 2649 through SEQ
ID NO: 2654.
[0074] The present invention also provides a substantially purified
antibody or fragment thereof, the antibody or fragment thereof
capable of specifically binding to a maize or a soybean
O-succinylhomoserine (thiol)-lyase enzyme or fragment thereof
encoded by a first nucleic acid molecule which specifically
hybridizes to a second nucleic acid molecule, the second nucleic
acid molecule having a nucleic acid sequence selected from the
group consisting of a complement of SEQ ID NO: 905 through SEQ ID
NO: 953 and SEQ ID NO: 2655 through SEQ ID NO: 2660 or a
substantially purified maize or soybean O-succinylhomoserine
(thiol)-lyase enzyme or fragment thereof encoded by a nucleic acid
sequence selected from the group consisting of SEQ ID NO: 905
through SEQ ID NO: 953 and SEQ ID NO: 2655 through SEQ ID NO:
2660.
[0075] The present invention also provides a substantially purified
antibody or fragment thereof, the antibody or fragment thereof
capable of specifically binding to a maize or a soybean
cystathionine .beta.-lyase enzyme or fragment thereof encoded by a
first nucleic acid molecule which specifically hybridizes to a
second nucleic acid molecule, the second nucleic acid molecule
having a nucleic acid sequence selected from the group consisting
of a complement of SEQ ID NO: 954 through SEQ ID NO: 963 and SEQ ID
NO: 2661 through SEQ ID NO: 2665 or a substantially purified maize
or soybean cystathionine .beta.-lyase enzyme or fragment thereof
encoded by a nucleic acid sequence selected from the group
consisting of SEQ ID NO: 954 through SEQ ID NO: 963 and SEQ ID NO:
2661 through SEQ ID NO: 2665.
[0076] The present invention also provides a substantially purified
antibody or fragment thereof, the antibody or fragment thereof
capable of specifically binding to a maize or a soybean
5-methyltetrahydropteroyltriglutamate-homocysteine-S-methyltransferase
enzyme or fragment thereof encoded by a first nucleic acid molecule
which specifically hybridizes to a second nucleic acid molecule,
the second nucleic acid molecule having a nucleic acid sequence
selected from the group consisting of a complement of SEQ ID NO:
964 through SEQ ID NO: 1353 and SEQ ID NO: 2666 through SEQ ID NO:
2992 or a substantially purified maize or soybean
5-methyltetrahydropteroyltriglutamate-homocysteine-S-methyltransferase
enzyme or fragment thereof encoded by a nucleic acid sequence
selected from the group consisting of SEQ ID NO: 964 through SEQ ID
NO: 1353 and SEQ ID NO: 2666 through SEQ ID NO: 2992.
[0077] The present invention also provides a substantially purified
antibody or fragment thereof, the antibody or fragment thereof
capable of specifically binding to a maize or a soybean
adenosylhomocyteinase enzyme or fragment thereof encoded by a first
nucleic acid molecule which specifically hybridizes to a second
nucleic acid molecule, the second nucleic acid molecule having a
nucleic acid sequence selected from the group consisting of a
complement of SEQ ID NO: 1354 through SEQ ID NO: 1630 and SEQ ID
NO: 2993 through SEQ ID NO: 3199 or a substantially purified maize
or soybean adenosylhomocyteinase enzyme or fragment thereof encoded
by a nucleic acid sequence selected from the group consisting of
SEQ ID NO: 1354 through SEQ ID NO: 1630 and SEQ ID NO: 2993 through
SEQ ID NO: 3199.
[0078] The present invention also provides a substantially purified
antibody or fragment thereof, the antibody or fragment thereof
capable of specifically binding to a maize or a soybean
cystathionine .beta.-synthase enzyme or fragment thereof encoded by
a first nucleic acid molecule which specifically hybridizes to a
second nucleic acid molecule, the second nucleic acid molecule
having a nucleic acid sequence selected from the group consisting
of a complement of SEQ ID NO: 1631 through SEQ ID NO: 1632 or a
substantially purified maize or soybean cystathionine
.beta.-synthase enzyme or fragment thereof encoded by a nucleic
acid sequence selected from the group consisting of SEQ ID NO: 1631
through SEQ ID NO: 1632.
[0079] The present invention also provides a substantially purified
antibody or fragment thereof, the antibody or fragment thereof
capable of specifically binding to a maize or a soybean
cystathionine .gamma.-lyase enzyme or fragment thereof encoded by a
first nucleic acid molecule which specifically hybridizes to a
second nucleic acid molecule, the second nucleic acid molecule
having a nucleic acid sequence selected from the group consisting
of a complement of SEQ ID NO: 1633 through SEQ ID NO: 1634 and SEQ
ID NO: 3203 through SEQ ID NO: 3204 or a substantially purified
maize or soybean cystathionine .gamma.-lyase enzyme or fragment
thereof encoded by a nucleic acid sequence selected from the group
consisting of SEQ ID NO: 1633 through SEQ ID NO: 1634 and SEQ ID
NO: 3203 through SEQ ID NO: 3204.
[0080] The present invention also provides a substantially purified
antibody or fragment thereof, the antibody or fragment thereof
capable of specifically binding to a maize or a soybean
O-acetylhomoserine enzyme or fragment thereof encoded by a first
nucleic acid molecule which specifically hybridizes to a second
nucleic acid molecule, the second nucleic acid molecule having a
nucleic acid sequence selected from the group consisting of a
complement of SEQ ID NO: 3200 through SEQ ID NO: 3202 or a
substantially purified maize or soybean O-acetylhomoserine enzyme
or fragment thereof encoded by a nucleic acid sequence selected
from the group consisting of SEQ ID NO: 3200 through SEQ ID NO:
3202.
[0081] The present invention also provides a transformed plant
having a nucleic acid molecule which comprises: (A) an exogenous
promoter region which functions in a plant cell to cause the
production of a mRNA molecule; (B) a structural nucleic acid
molecule comprising a nucleic acid sequence selected from the group
consisting of (a) a nucleic acid sequence which encodes for
methionine adenosyltransferase or fragment thereof; (b) a nucleic
acid sequence which encodes for S-adenosyl-methionine decarboxylase
or fragment thereof; (c) a nucleic acid sequence which encodes for
aspartate kinase or fragment thereof; (d) a nucleic acid sequence
which encodes for aspartate-semialdehyde dehydrogenase or fragment
thereof; (e) a nucleic acid sequence which encodes for
cystathionine gamma-synthase or a fragment thereof; (f) a nucleic
acid sequence which encodes for cystathionine beta-lysase or a
fragment thereof; (g) a nucleic acid sequence which encodes for
5-methyltetrahydropteroyl-triglutamate-homocysteine-S-methyltransferase
or a fragment thereof; and (h) a nucleic acid sequence which is
complementary to any of the nucleic acid sequences of (a) through
(g); and (C) a 3' non-translated sequence that functions in said
plant cell to cause termination of transcription and addition of
polyadenylated ribonucleotides to a 3' end of said mRNA
molecule.
[0082] The present invention also provides a transformed plant
having a nucleic acid molecule which comprises: (A) an exogenous
promoter region which functions in a plant cell to cause the
production of a mRNA molecule; which is linked to (B) a structural
nucleic acid molecule, wherein the structural nucleic acid molecule
encodes a plant methionine pathway enzyme or fragment thereof, the
structural nucleic acid molecule comprising a nucleic acid sequence
selected from the group consisting of SEQ ID NO: 1 through SEQ ID
NO: 3204 or fragment thereof; which is linked to (C) a 3'
non-translated sequence that functions in the plant cell to cause
termination of transcription and addition of polyadenylated
ribonucleotides to a 3' end of the mRNA molecule.
[0083] The present invention also provides a transformed plant
having a nucleic acid molecule which comprises: (A) an exogenous
promoter region which functions in a plant cell to cause the
production of a mRNA molecule; which is linked to (B) a structural
nucleic acid molecule, wherein the structural nucleic acid molecule
is selected from the group consisting of a nucleic acid molecule
that encodes a maize or a soybean methionine adenosyltransferase
enzyme or fragment thereof, a nucleic acid molecule that encodes a
maize or a soybean S-adenosylmethionine decarboxylase enzyme or
fragment thereof, a nucleic acid molecule that encodes a maize or
soybean aspartate kinase enzyme or fragment thereof, a nucleic acid
molecule that encodes a maize or a soybean aspartate-semialdehyde
dehydrogenase enzyme or fragment thereof, a nucleic acid molecule
that encodes a maize or a soybean O-succinylhomoserine
(thiol)-lyase enzyme or fragment thereof, a nucleic acid molecule
that encodes a maize or a soybean cystathionine .beta.-lyase enzyme
or fragment thereof, a nucleic acid molecule that encodes a maize
or a soybean
5-methyltetrahydropteroyltriglutamate-homocysteine-S-methyltransferase
enzyme or fragment thereof, a nucleic acid molecule that encodes a
maize or a soybean adenosylhomocysteinase enzyme or fragment
thereof, a nucleic acid molecule that encodes a maize or a soybean
cystathionine .beta.-synthase enzyme or fragment thereof, a nucleic
acid molecule that encodes a maize or a soybean cystathionine
.gamma.-lyase enzyme or fragment thereof and a nucleic acid
molecule that encodes a maize or a soybean O-acetylhomoserine
(thiol)-lyase enzyme or fragment thereof; which is linked to (C) a
3' non-translated sequence that functions in the plant cell to
cause termination of transcription and addition of polyadenylated
ribonucleotides to a 3' end of the mRNA molecule.
[0084] The present invention also provides a transformed plant
having a nucleic acid molecule which comprises: (A) an exogenous
promoter region which functions in a plant cell to cause the
production of a mRNA molecule; which is linked to (B) a transcribed
nucleic acid molecule with a transcribed strand and a
non-transcribed strand, wherein the transcribed strand is
complementary to a nucleic acid molecule comprising a nucleic acid
sequence selected from the group consisting of SEQ ID NO: 1 through
SEQ ID NO: 3204 or fragment thereof; which is linked to (C) a 3'
non-translated sequence that functions in plant cells to cause
termination of transcription and addition of polyadenylated
ribonucleotides to a 3' end of the mRNA molecule.
[0085] The present invention also provides a transformed plant
having a nucleic acid molecule which comprises: (A) an exogenous
promoter region which functions in a plant cell to cause the
production of a mRNA molecule; which is linked to: (B) a
transcribed nucleic acid molecule with a transcribed strand and a
non-transcribed strand, wherein a transcribed mRNA of the
transcribed strand is complementary to an endogenous mRNA molecule
having a nucleic acid sequence selected from the group consisting
of an endogenous mRNA molecule that encodes a maize or a soybean
methionine adenosyltransferase enzyme or fragment thereof, an
endogenous mRNA molecule that encodes a maize or a soybean
S-adenosylmethionine decarboxylase enzyme or fragment thereof, an
endogenous mRNA molecule that encodes a maize or a soybean
aspartate kinase enzyme or fragment thereof, an endogenous mRNA
molecule that encodes a maize or a soybean aspartate-semialdehyde
dehydrogenase enzyme or fragment thereof, an endogenous mRNA
molecule that encodes a maize or a soybean O-succinylhomoserine
(thiol)-lyase enzyme or fragment thereof, an endogenous mRNA
molecule that encodes a maize or a soybean cystathionine
.beta.-lyase enzyme or fragment thereof, an endogenous mRNA
molecule that encodes a maize or a soybean
5-methyltetrahydropteroyltriglutamate-homocysteine-S-methyltransferase
enzyme or fragment thereof, an endogenous mRNA molecule that
encodes a maize or a soybean adenosylhomocysteinase enzyme or
fragment thereof, an endogenous mRNA molecule that encodes a maize
or a soybean cystathionine .beta.-synthase enzyme or fragment
thereof, an endogenous mRNA molecule that encodes a maize or a
soybean cystathionine .gamma.-lyase enzyme or fragment thereof and
an endogenous mRNA molecule that encodes a maize or a soybean
O-acetylhomoserine (thiol)-lyase enzyme or fragment thereof; which
is linked to (C) a 3' non-translated sequence that functions in the
plant cell to cause termination of transcription and addition of
polyadenylated ribonucleotides to a 3' end of the mRNA
molecule.
[0086] The present invention also provides a method for determining
a level or pattern in a plant cell of an enzyme in a plant
metabolic pathway comprising: (A) incubating, under conditions
permitting nucleic acid hybridization, a marker nucleic acid
molecule, said marker nucleic acid molecule selected from the group
of marker nucleic acid molecules which specifically hybridize to a
nucleic acid molecule having the nucleic acid sequence of SEQ ID
NO: 1 through SEQ ID NO: 3204 or compliments thereof, with a
complementary nucleic acid molecule obtained from said plant cell
or plant tissue, wherein nucleic acid hybridization between said
marker nucleic acid molecule and said complementary nucleic acid
molecule obtained from said plant cell or plant tissue permits the
detection of an mRNA for said enzyme; (B) permitting hybridization
between said marker nucleic acid molecule and said complementary
nucleic acid molecule obtained from said plant cell or plant
tissue; and (C) detecting the level or pattern of said
complementary nucleic acid, wherein the detection of said
complementary nucleic acid is predictive of the level or pattern of
said enzyme in said plant metabolic pathway.
[0087] The present invention also provides a method for determining
a level or pattern of a plant methionine pathway enzyme in a plant
cell or plant tissue comprising: (A) incubating, under conditions
permitting nucleic acid hybridization, a marker nucleic acid
molecule, the marker nucleic acid molecule having a nucleic acid
sequence selected from the group consisting of SEQ ID NO: 1 through
SEQ ID NO: 3204 or complement thereof or fragment of either, with a
complementary nucleic acid molecule obtained from the plant cell or
plant tissue, wherein nucleic acid hybridization between the marker
nucleic acid molecule and the complementary nucleic acid molecule
obtained from the plant cell or plant tissue permits the detection
of the plant methionine pathway enzyme; (B) permitting
hybridization between the marker nucleic acid molecule and the
complementary nucleic acid molecule obtained from the plant cell or
plant tissue; and (C) detecting the level or pattern of the
complementary nucleic acid, wherein the detection of the
complementary nucleic acid is predictive of the level or pattern of
the plant methionine pathway enzyme.
[0088] The present invention also provides a method for determining
a level or pattern of a plant methionine pathway enzyme in a plant
cell or plant tissue comprising: (A) incubating, under conditions
permitting nucleic acid hybridization, a marker nucleic acid
molecule, the marker nucleic acid molecule comprising a nucleic
acid molecule that encodes a maize or a soybean methionine
adenosyltransferase enzyme or complement thereof or fragment of
either, a nucleic acid molecule that encodes a maize or a soybean
S-adenosylmethionine decarboxylase enzyme or complement thereof or
fragment of either, a nucleic acid molecule that encodes a maize or
a soybean aspartate kinase enzyme or complement thereof or fragment
of either, a nucleic acid molecule that encodes a maize or a
soybean aspartate-semialdehyde dehydrogenase enzyme or complement
thereof or fragment of either, a nucleic acid molecule that encodes
a maize or a soybean O-succinylhomoserine (thiol)-lyase enzyme or
complement thereof or fragment of either, a nucleic acid molecule
that encodes a maize or a soybean cystathionine .beta.-lyase enzyme
or complement thereof or fragment of either, a nucleic acid
molecule that encodes a maize or a soybean
5-methyltetrahydropteroyltriglutamate-homocysteine-S-methyltransferase
enzyme or complement thereof or fragment of either, a nucleic acid
molecule that encodes a maize or a soybean adenosylhomocysteinase
enzyme or complement thereof or fragment of either, a nucleic acid
molecule that encodes a maize or a soybean cystathionine
.beta.-synthase enzyme or complement thereof or fragment of either,
a nucleic acid molecule that encodes a maize or a soybean
cytsathionine .gamma.-lyase enzyme or complement thereof or
fragment of either and a nucleic acid molecule that encodes a maize
or a soybean O-acetylhomoserine (thiol)-lyase enzyme or complement
thereof or fragment of either, with a complementary nucleic acid
molecule obtained from the plant cell or plant tissue, wherein
nucleic acid hybridization between the marker nucleic acid molecule
and the complementary nucleic acid molecule obtained from the plant
cell or plant tissue permits the detection of the plant methionine
pathway enzyme; (B) permitting hybridization between the marker
nucleic acid molecule and the complementary nucleic acid molecule
obtained from the plant cell or plant tissue; and (C) detecting the
level or pattern of the complementary nucleic acid, wherein the
detection of the complementary nucleic acid is predictive of the
level or pattern of the plant methionine pathway enzyme.
[0089] The present invention also provides a method for determining
a level or pattern of a plant methionine pathway enzyme in a plant
cell or plant tissue under evaluation which comprises assaying the
concentration of a molecule, whose concentration is dependent upon
the expression of a gene, the gene specifically hybridizes to a
nucleic acid molecule having a nucleic acid sequence selected from
the group consisting of SEQ ID NO: 1 through SEQ ID NO: 3204 or
complements thereof, in comparison to the concentration of that
molecule present in a reference plant cell or a reference plant
tissue with a known level or pattern of the plant methionine
pathway enzyme, wherein the assayed concentration of the molecule
is compared to the assayed concentration of the molecule in the
reference plant cell or reference plant tissue with the known level
or pattern of the plant methionine pathway enzyme.
[0090] The present invention also provides a method for determining
a level or pattern of a plant methionine pathway enzyme in a plant
cell or plant tissue under evaluation which comprises assaying the
concentration of a molecule, whose concentration is dependent upon
the expression of a gene, the gene specifically hybridizes to a
nucleic acid molecule selected from the group consisting of a
nucleic acid molecule that encodes a maize or a soybean methionine
adenosyltransferase enzyme or complement thereof, a nucleic acid
molecule that encodes a maize or a soybean S-adenosylmethionine
decarboxylase enzyme or complement thereof, a nucleic acid molecule
that encodes a maize or a soybean aspartate kinase enzyme or
complement thereof, a nucleic acid molecule that encodes a maize or
a soybean aspartate-semialdehyde dehydrogenase enzyme or complement
thereof, a nucleic acid molecule that encodes a maize or a soybean
O-succinylhomoserine (thiol)-lyase enzyme or complement thereof, a
nucleic acid molecule that encodes a maize or a soybean
cystathionine .beta.-lyase enzyme or complement thereof, a nucleic
acid molecule that encodes a maize or a soybean
5-methyltetrahydropteroyltriglutamate-homocysteine-S-methyltransferase
enzyme or complement thereof, a nucleic acid molecule that encodes
a maize or a soybean adenosylhomocyteinase enzyme or complement
thereof, a nucleic acid molecule that encodes a maize or a soybean
cystathionine .beta.-synthase enzyme or complement thereof, a
nucleic acid molecule that encodes a maize or a soybean
cystathionine .gamma.-lyase enzyme or complement thereof and a
nucleic acid molecule that encodes a maize or a soybean
O-acetylhomoserine (thiol)-lyase enzyme or complement thereof, in
comparison to the concentration of that molecule present in a
reference plant cell or a reference plant tissue with a known level
or pattern of the plant methionine pathway enzyme, wherein the
assayed concentration of the molecule is compared to the assayed
concentration of the molecule in the reference plant cell or the
reference plant tissue with the known level or pattern of the plant
methionine pathway enzyme.
[0091] A method of determining a mutation in a plant whose presence
is predictive of a mutation affecting a level or pattern of a
protein comprising the steps: (A) incubating, under conditions
permitting nucleic acid hybridization, a marker nucleic acid, said
marker nucleic acid selected from the group of marker nucleic acid
molecules which specifically hybridize to a nucleic acid molecule
having a nucleic acid sequence selected from the group of SEQ ID
NO: 1 through SEQ ID NO: 3204 or complements thereof and a
complementary nucleic acid molecule obtained from said plant,
wherein nucleic acid hybridization between said marker nucleic acid
molecule and said complementary nucleic acid molecule obtained from
said plant permits the detection of a polymorphism whose presence
is predictive of a mutation affecting said level or pattern of said
plant methionine pathway enzyme in said plant; (B) permitting
hybridization between said marker nucleic acid molecule and said
complementary nucleic acid molecule obtained from said plant; and
(C) detecting the presence of said polymorphism, wherein the
detection of said polymorphism is predictive of said mutation.
[0092] The present invention also provides a method for determining
a mutation in a plant whose presence is predictive of a mutation
affecting the level or pattern of a plant methionine pathway enzyme
comprising the steps: (A) incubating, under conditions permitting
nucleic acid hybridization, a marker nucleic acid molecule, the
marker nucleic acid molecule comprising a nucleic acid molecule
that is linked to a gene, the gene specifically hybridizes to a
nucleic acid molecule having a nucleic acid sequence selected from
the group consisting of SEQ ID NO: 1 through SEQ ID NO: 3204 or
complements thereof and a complementary nucleic acid molecule
obtained from the plant, wherein nucleic acid hybridization between
the marker nucleic acid molecule and the complementary nucleic acid
molecule obtained from the plant permits the detection of a
polymorphism whose presence is predictive of a mutation affecting
the level or pattern of the plant methionine pathway enzyme in the
plant; (B) permitting hybridization between the marker nucleic acid
molecule and the complementary nucleic acid molecule obtained from
the plant; and (C) detecting the presence of the polymorphism,
wherein the detection of the polymorphism is predictive of the
mutation.
[0093] The present invention also provides a method for determining
a mutation in a plant whose presence is predictive of a mutation
affecting the level or pattern of a plant methionine pathway enzyme
comprising the steps: (A) incubating, under conditions permitting
nucleic acid hybridization, a marker nucleic acid molecule, the
marker nucleic acid molecule comprising a nucleic acid molecule
that is linked to a gene, the gene specifically hybridizes to a
nucleic acid molecule selected from the group consisting of a
nucleic acid molecule that encodes a maize or a soybean methionine
adenosyltransferase enzyme or complement thereof, a nucleic acid
molecule that encodes a maize or a soybean S-adenosylmethionine
decarboxylase enzyme or complement thereof, a nucleic acid molecule
that encodes a maize or a soybean aspartate kinase enzyme or
complement thereof, a nucleic acid molecule that encodes a maize or
a soybean aspartate-semialdehyde dehydrogenase enzyme or complement
thereof, a nucleic acid molecule that encodes a maize or a soybean
O-succinylhomoserine (thiol)-lyase enzyme or complement thereof, a
nucleic acid molecule that encodes a maize or a soybean
cystathionine .beta.-lyase enzyme or complement thereof, a nucleic
acid molecule that encodes a maize or a soybean
5-methyltetrahydropteroyltriglutamate-homocysteine-S-methyltransferase
enzyme or complement thereof, a nucleic acid molecule that encodes
a maize or a soybean adenosylhomocyteinase enzyme or complement
thereof, a nucleic acid molecule that encodes a maize or a soybean
cystathionine .beta.-synthase enzyme or complement thereof, a
nucleic acid molecule that encodes a maize or a soybean
cystathionine .gamma.-lyase enzyme or complement thereof and a
nucleic acid molecule that encodes a maize or a soybean
O-acetylhomoserine (thiol)-lyase enzyme or complement thereof and a
complementary nucleic acid molecule obtained from the plant,
wherein nucleic acid hybridization between the marker nucleic acid
molecule and the complementary nucleic acid molecule obtained from
the plant permits the detection of a polymorphism whose presence is
predictive of a mutation affecting the level or pattern of the
plant methionine pathway enzyme in the plant; (B) permitting
hybridization between the marker nucleic acid molecule and the
complementary nucleic acid molecule obtained from the plant; and
(C) detecting the presence of the polymorphism, wherein the
detection of the polymorphism is predictive of the mutation.
[0094] A method of producing a plant containing an overexpressed
protein comprising: (A) transforming said plant with a functional
nucleic acid molecule, wherein the functional nucleic acid molecule
comprises a promoter region, wherein said promoter region is linked
to a structural region, wherein said structural region has a
nucleic acid sequence selected from group consisting of SEQ ID NO:
1 through SEQ ID NO: 3204 wherein said structural region is linked
to a 3' non-translated sequence that functions in the plant to
cause termination of transcription and addition of polyadenylated
ribonucleotides to a 3' end of a mRNA molecule; and wherein said
functional nucleic acid molecule results in overexpression of the
protein; and (B) growing said transformed plant.
[0095] The present invention also provides a method of producing a
plant containing an overexpressed plant methionine enzyme
comprising: (A) transforming the plant with a functional nucleic
acid molecule, wherein the functional nucleic acid molecule
comprises a promoter region, wherein the promoter region is linked
to a structural region, wherein the structural region comprises a
nucleic acid molecule having a nucleic acid sequence selected from
the group consisting of SEQ ID NO: 1 through SEQ ID NO: 3204 or
fragment thereof; wherein the structural region is linked to a 3'
non-translated sequence that functions in the plant to cause
termination of transcription and addition of polyadenylated
ribonucleotides to a 3' end of a mRNA molecule; and wherein the
functional nucleic acid molecule results in overexpression of the
plant methionine pathway enzyme; and (B) growing the transformed
plant.
[0096] The present invention also provides a method of producing a
plant containing an overexpressed plant methionine pathway enzyme
comprising: (A) transforming the plant with a functional nucleic
acid molecule, wherein the functional nucleic acid molecule
comprises a promoter region, wherein the promoter region is linked
to a structural region, wherein the structural region comprises a
nucleic acid molecule selected from the group consisting of a
nucleic acid molecule that encodes a maize or a soybean methionine
adenosyltransferase enzyme or fragment thereof, a nucleic acid
molecule that encodes a maize or a soybean S-adenosylmethionine
decarboxylase enzyme or fragment thereof, a nucleic acid molecule
that encodes a maize or a soybean aspartate kinase enzyme or
fragment thereof, a nucleic acid molecule that encodes a maize or a
soybean aspartate-semialdehyde dehydrogenase enzyme or fragment
thereof, a nucleic acid molecule that encodes a maize or a soybean
O-succinylhomoserine (thiol)-lyase enzyme or fragment thereof, a
nucleic acid molecule that encodes a maize or a soybean
cystathionine .beta.-lyase enzyme or fragment thereof, a nucleic
acid molecule that encodes a maize or a soybean
5-methyltetrahydropteroyltriglutamate-homocysteine-S-methyltransferase
enzyme or fragment thereof, a nucleic acid molecule that encodes a
maize or a soybean adenosylhomocysteinase enzyme or fragment
thereof, a nucleic acid molecule that encodes a maize or a soybean
cystathionine .beta.-synthase enzyme or fragment thereof, a nucleic
acid molecule that encodes a maize or a soybean cystathionine
.gamma.-lyase enzyme or fragment thereof and a nucleic acid
molecule that encodes a maize or a soybean O-acetylhomoserine
(thiol)-lyase enzyme or fragment thereof; wherein the structural
region is linked to a 3' non-translated sequence that functions in
the plant to cause termination of transcription and addition of
polyadenylated ribonucleotides to a 3' end of a mRNA molecule; and
wherein the functional nucleic acid molecule results in
overexpression of the plant methionine pathway enzyme protein; and
(B) growing the transformed plant.
[0097] The present invention also provides a method of producing a
plant containing reduced levels of a plant methionine pathway
enzyme comprising: (A) transforming the plant with a functional
nucleic acid molecule, wherein the functional nucleic acid molecule
comprises a promoter region, wherein the promoter region is linked
to a structural region, wherein the structural region comprises a
nucleic acid molecule having a nucleic acid sequence selected from
the group consisting of SEQ ID NO: 1 through SEQ ID NO: 3204;
wherein the structural region is linked to a 3' non-translated
sequence that functions in the plant to cause termination of
transcription and addition of polyadenylated ribonucleotides to a
3' end of a mRNA molecule; and wherein the functional nucleic acid
molecule results in co-suppression of the plant methionine pathway
enzyme protein; and (B) growing the transformed plant.
[0098] The present invention also provides a method of producing a
plant containing reduced levels of a plant methionine pathway
enzyme comprising: (A) transforming the plant with a functional
nucleic acid molecule, wherein the functional nucleic acid molecule
comprises a promoter region, wherein the promoter region is linked
to a structural region, wherein the structural region comprises a
nucleic acid molecule having a nucleic acid sequence selected from
the group consisting of a nucleic acid molecule that encodes a
maize or a soybean methionine adenosyltransferase enzyme or
fragment thereof, a nucleic acid molecule that encodes a maize or a
soybean S-adenosylmethionine decarboxylase enzyme or fragment
thereof, a nucleic acid molecule that encodes a maize or a soybean
aspartate kinase enzyme or fragment thereof, a nucleic acid
molecule that encodes a maize or a soybean aspartate-semialdehyde
dehydrogenase enzyme or fragment thereof, a nucleic acid molecule
that encodes a maize or a soybean O-succinylhomoserine
(thiol)-lyase enzyme or fragment thereof, a nucleic acid molecule
that encodes a maize or a soybean cystathionine .beta.-lyase enzyme
or fragment thereof, a nucleic acid molecule that encodes a maize
or a soybean
5-methyltetrahythopteroyltriglutamate-homocysteine-S-methyltransferase
enzyme or fragment thereof, a nucleic acid molecule that encodes a
maize or a soybean adenosylhomocysteinase enzyme or fragment
thereof, a nucleic acid molecule that encodes a maize or a soybean
cystathionine .beta.-synthase enzyme or fragment thereof, a nucleic
acid molecule that encodes a maize or a soybean cystathionine
.gamma.-lyase enzyme or fragment thereof and a nucleic acid
molecule that encodes a maize or a soybean O-acetylhomoserine
(thiol)-lyase enzyme or fragment thereof; wherein the structural
region is linked to a 3' non-translated sequence that functions in
the plant to cause termination of transcription and addition of
polyadenylated ribonucleotides to a 3' end of a mRNA molecule; and
wherein the functional nucleic acid molecule results in
co-suppression of the plant methionine pathway enzyme; and (B)
growing the transformed plant.
[0099] The present invention also provides a method for reducing
expression of a plant methionine pathway enzyme in a plant
comprising: (A) transforming the plant with a nucleic acid
molecule, the nucleic acid molecule having an exogenous promoter
region which functions in a plant cell to cause the production of a
mRNA molecule, wherein the exogenous promoter region is linked to a
transcribed nucleic acid molecule having a transcribed strand and a
non-transcribed strand, wherein the transcribed strand is
complementary to a nucleic acid molecule having a nucleic acid
sequence selected from the group consisting of SEQ ID NO: 1 through
SEQ ID NO: 3204 or complements thereof or fragments of either and
the transcribed strand is complementary to an endogenous mRNA
molecule; and wherein the transcribed nucleic acid molecule is
linked to a 3' non-translated sequence that functions in the plant
cell to cause termination of transcription and addition of
polyadenylated ribonucleotides to a 3' end of a mRNA molecule; and
(B) growing the transformed plant.
[0100] The present invention also provides a method for reducing
expression of a plant methionine pathway enzyme in a plant
comprising: (A) transforming the plant with a nucleic acid
molecule, the nucleic acid molecule having an exogenous promoter
region which functions in a plant cell to cause the production of a
mRNA molecule, wherein the exogenous promoter region is linked to a
transcribed nucleic acid molecule having a transcribed strand and a
non-transcribed strand, wherein a transcribed mRNA of the
transcribed strand is complementary to a nucleic acid molecule
selected from the group consisting of an endogenous mRNA molecule
that encodes a maize or a soybean methionine adenosyltransferase
enzyme or fragment thereof, an endogenous mRNA molecule that
encodes a maize or a soybean S-adenosylmethionine decarboxylase
enzyme or fragment thereof, an endogenous mRNA molecule that
encodes a maize or a soybean aspartate kinase enzyme or fragment
thereof, an endogenous mRNA molecule that encodes a maize or a
soybean aspartate-semialdehyde dehydrogenase enzyme or fragment
thereof, an endogenous mRNA molecule that encodes a maize or a
soybean O-succinylhomoserine (thiol)-lyase enzyme or fragment
thereof, an endogenous mRNA molecule that encodes a maize or a
soybean cystathionine .beta.-lyase enzyme or fragment thereof, an
endogenous mRNA molecule that encodes a maize or a soybean
5-methyltetrahydropteroyltriglutamate-homocysteine-S-methyltransferase
enzyme or fragment thereof, an endogenous mRNA molecule that
encodes a maize or a soybean adenosylhomocysteinase enzyme or
fragment thereof, an endogenous mRNA molecule that encodes a maize
or a soybean cystathionine f-.beta.-synthase enzyme or fragment
thereof, an endogenous mRNA molecule that encodes a maize or a
soybean cystathionine .gamma.-lyase enzyme or fragment thereof and
an endogenous mRNA molecule that encodes a maize or a soybean
O-acetylhomoserine (thiol)-lyase enzyme or fragment thereof; and
wherein the transcribed nucleic acid molecule is linked to a 3'
non-translated sequence that functions in the plant cell to cause
termination of transcription and addition of polyadenylated
ribonucleotides to a 3' end of a mRNA molecule; and (B) growing the
transformed plant.
[0101] The present invention also provides a method of determining
an association between a polymorphism and a plant trait comprising:
(A) hybridizing a nucleic acid molecule specific for the
polymorphism to genetic material of a plant, wherein the nucleic
acid molecule has a nucleic acid sequence selected from the group
consisting of SEQ ID NO: 1 through SEQ ID NO: 3204 or complements
thereof or fragment thereof; and (B) calculating the degree of
association between the polymorphism and the plant trait.
[0102] The present invention also provides a method of determining
an association between a polymorphism and a plant trait comprising:
(A) hybridizing a nucleic acid molecule specific for the
polymorphism to genetic material of a plant, wherein the nucleic
acid molecule is selected from the group consisting of a nucleic
acid molecule that encodes a maize or a soybean methionine
adenosyltransferase enzyme complement thereof or fragment of
either, a nucleic acid molecule that encodes a maize or a soybean
S-adenosylmethionine decarboxylase enzyme complement thereof or
fragment of either, a nucleic acid molecule that encodes a maize or
a soybean aspartate kinase enzyme complement thereof or fragment of
either, a nucleic acid molecule that encodes a maize or a soybean
aspartate-semialdehyde dehydrogenase enzyme complement thereof or
fragment of either, a nucleic acid molecule that encodes a maize or
a soybean O-succinylhomoserine (thiol)-lyase enzyme complement
thereof or fragment of either, a nucleic acid molecule that encodes
a maize or a soybean cystathionine .beta.-lyase enzyme complement
thereof or fragment of either, a nucleic acid molecule that encodes
a maize or a soybean
5-methyltetrahydropteroyltriglutamate-homocysteine-S-methyltransferase
enzyme or complement thereof or fragment of either; a nucleic acid
molecule that encodes a maize or a soybean adenosylhomocysteinase
enzyme or complement thereof or fragment of either, a nucleic acid
molecule that encodes a maize or a soybean cystathionine
.beta.-synthase enzyme or complement thereof or fragment of either,
a nucleic acid molecule that encodes a maize or a soybean
cytsathionine .gamma.-lyase enzyme or complement thereof or
fragment of either and a nucleic acid molecule that encodes a maize
or a soybean O-acetylhomoserine (thiol)-lyase enzyme or complement
thereof or fragment of either and (B) calculating the degree of
association between the polymorphism and the plant trait.
[0103] The present invention also provides a method of isolating a
nucleic acid that encodes a plant methionine pathway enzyme or
fragment thereof comprising: (A) incubating under conditions
permitting nucleic acid hybridization, a first nucleic acid
molecule comprising a nucleic acid sequence selected from the group
consisting of SEQ ID NO: 1 through SEQ ID NO: 3204 or complements
thereof or fragment of either with a complementary second nucleic
acid molecule obtained from a plant cell or plant tissue; (B)
permitting hybridization between the first nucleic acid molecule
and the second nucleic acid molecule obtained from the plant cell
or plant tissue; and (C) isolating the second nucleic acid
molecule.
[0104] The present invention also provides a method of isolating a
nucleic acid molecule that encodes a plant methionine pathway
enzyme or fragment thereof comprising: (A) incubating under
conditions permitting nucleic acid hybridization, a first nucleic
acid molecule selected from the group consisting of a nucleic acid
molecule that encodes a maize or a soybean methionine
adenosyltransferase enzyme complement thereof or fragment of
either, a nucleic acid molecule that encodes a maize or a soybean
S-adenosylmethionine decarboxylase enzyme or complement thereof or
fragment of either, a nucleic acid molecule that encodes a maize or
a soybean aspartate kinase enzyme complement thereof or fragment of
either, a nucleic acid molecule that encodes a maize or a soybean
aspartate-semialdehyde dehydrogenase enzyme or complement thereof
or fragment of either, a nucleic acid molecule that encodes a maize
or a soybean O-succinylhomoserine (thiol)-lyase enzyme or
complement thereof or fragment of either and a nucleic acid
molecule that encodes a maize or a soybean cystathionine
.beta.-lyase enzyme or complement thereof or fragment of either, a
nucleic acid molecule that encodes a maize or a soybean
5-methyltetrahydropteroyltriglutamate-homocysteine-S-methyltransf-
erase enzyme or complement thereof or fragment of either, a nucleic
acid molecule that encodes a maize or a soybean
adenosylhomocysteinase enzyme or complement thereof or fragment of
either, a nucleic acid molecule that encodes a maize or a soybean
cystathionine .beta.-synthase enzyme or complement thereof or
fragment of either, a nucleic acid molecule that encodes a maize or
a soybean cytsathionine .gamma.-lyase enzyme or complement thereof
or fragment of either and a nucleic acid molecule that encodes a
maize or a soybean O-acetylhomoserine (thiol)-lyase enzyme or
complement thereof or fragment of either, with a complementary
second nucleic acid molecule obtained from a plant cell or plant
tissue; (B) permitting hybridization between the plant methionine
pathway nucleic acid molecule and the complementary nucleic acid
molecule obtained from the plant cell or plant tissue; and (C)
isolating the second nucleic acid molecule.
DETAILED DESCRIPTION OF THE INVENTION
Definitions and Agents of the Present Invention
Definitions
[0105] As used herein, a methionine pathway enzyme is any enzyme
that is associated with the synthesis or degradation of
methionine.
[0106] As used herein, a methionine synthesis enzyme is any enzyme
that is associated with the synthesis of methionine.
[0107] As used herein, a methionine degradation enzyme is any
enzyme that is associated with the degradation of methionine.
[0108] As used herein, methionine adenosyltransferase is any enzyme
that catalyzes the conversion of methionine to
S-adenosylmethionine.
[0109] As used herein, S-adenosylmethionine decarboxylase is any
enzyme that catalyzes the reaction that converts
S-adenosylmethionine to (5-deoxy-5-adenosyl)(3-aminopropyl)
methylsulfonuim salt.
[0110] As used herein, aspartate kinase is any enzyme that
catalyzes the conversion of aspartate to .beta.-aspartyl
phosphate.
[0111] As used herein, aspartate semialdehyde dehydrogenase is any
enzyme that catalyzes the conversion of .beta.-aspartyl phosphate
to aspartate-semialdehyde via an NADPH-dependent reaction.
[0112] As used herein, O-succinylhomoserine (thiol)-lyase refers to
any enzyme that catalyzes the conversion of O-phosphohomoserine to
and cysteine to cystathionine.
[0113] As used herein, cystathionine .beta.-lyase is any enzyme
that catalyzes the conversion of cystathionine to homocysteine,
pyruvate and ammonia.
[0114] As used herein,
5-methyltetrhydropterolytriglutamate-homocysteine
S-methyltransferase refers to any enzyme which catalyzes the
conversion of homocysteine via methylation to methionine.
[0115] As used herein, adenosylhomocysteinase refers to any enzyme
that catalyzes the ATP-dependent conversion of S-adenosylmethionine
(AdoMet) to methylthioadenosine and L-homoserine.
[0116] As used herein, cystathionine .beta.-synthase refers to any
enzyme that catalyzes the conversion of homocysteine and serine to
cystathionine.
[0117] As used herein, cystathionine .gamma.-lyase refers to any
enzyme that catalyzes the y cleavage of cystationine.
[0118] As used herein, O-acetyhomoserine (thiol)-lyase refers to
any enzyme that catalyzes the conversion of O-acetylhomoserine and
sulfur to homocysteine.
Agents
[0119] (a) Nucleic Acid Molecules
[0120] Agents of the present invention include plant nucleic acid
molecules and more specifically include maize and soybean nucleic
acid molecules and more specifically include nucleic acid molecules
of the maize genotypes B73 (Illinois Foundation Seeds, Champaign,
Ill. U.S.A.), B73.times.Mo17 (Illinois Foundation Seeds, Champaign,
Ill. U.S.A.), DK604 (Dekalb Genetics, Dekalb, Ill. U.S.A.), H99
(Illinois Foundation Seeds, Champaign, Ill. U.S.A.), RX601 (Asgrow
Seed Company, Des Moines, Iowa), Mo17 (Illinois Foundation Seeds,
Champaign, Ill. U.S.A.), and soybean types Asgrow 3244 (Asgrow Seed
Company, Des Moines, Iowa), C1944 (United States Department of
Agriculture (USDA) Soybean Germplasm Collection, Urbana, Ill.
U.S.A.), Cristalina (USDA Soybean Germplasm Collection, Urbana,
Ill. U.S.A.), FT108 (Monsoy, Brazil), Hartwig (USDA Soybean
Germplasm Collection, Urbana, Ill. U.S.A.), BW211S Null (Tohoku
University, Morioka, Japan), PI507354 (USDA Soybean Germplasm
Collection, Urbana, Ill. U.S.A.), Asgrow A4922 (Asgrow Seed
Company, Des Moines, Iowa U.S.A.), PI227687 (USDA Soybean Germplasm
Collection, Urbana, Ill. U.S.A.), PI229358 (USDA Soybean Germplasm
Collection, Urbana, Ill. U.S.A.) and Asgrow A3237 (Asgrow Seed
Company, Des Moines, Iowa U.S.A.).
[0121] A subset of the nucleic acid molecules of the present
invention includes nucleic acid molecules that are marker
molecules. Another subset of the nucleic acid molecules of the
present invention include nucleic acid molecules that encode a
protein or fragment thereof. Another subset of the nucleic acid
molecules of the present invention are EST molecules.
[0122] Fragment nucleic acid molecules may encode significant
portion(s) of, or indeed most of, these nucleic acid molecules.
Alternatively, the fragments may comprise smaller oligonucleotides
(having from about 15 to about 250 nucleotide residues and more
preferably, about 15 to about 30 nucleotide residues).
[0123] As used herein, an agent, be it a naturally occurring
molecule or otherwise may be "substantially purified," if desired,
such that one or more molecules that is or may be present in a
naturally occurring preparation containing that molecule will have
been removed or will be present at a lower concentration than that
at which it would normally be found.
[0124] The agents of the present invention will preferably be
"biologically active" with respect to either a structural
attribute, such as the capacity of a nucleic acid to hybridize to
another nucleic acid molecule, or the ability of a protein to be
bound by an antibody (or to compete with another molecule for such
binding). Alternatively, such an attribute may be catalytic and
thus involve the capacity of the agent to mediate a chemical
reaction or response.
[0125] The agents of the present invention may also be recombinant.
As used herein, the term recombinant means any agent (e.g. DNA,
peptide etc.), that is, or results, however indirect, from human
manipulation of a nucleic acid molecule.
[0126] It is understood that the agents of the present invention
may be labeled with reagents that facilitate detection of the agent
(e.g. fluorescent labels, Prober et al., Science 238:336-340
(1987); Albarella et al., EP 144914; chemical labels, Sheldon et
al., U.S. Pat. No. 4,582,789; Albarella et al., U.S. Pat. No.
4,563,417; modified bases, Miyoshi et al., EP 119448, all of which
are hereby incorporated by reference in their entirety).
[0127] It is further understood, that the present invention
provides recombinant bacterial, mammalian, microbial, insect,
fungal and plant cells and viral constructs comprising the agents
of the present invention. (See, for example, Uses of the Agents of
the Invention, Section (a) Plant Constructs and Plant
Transformants; Section (b) Fungal Constructs and Fungal
Transformants; Section (c) Mammalian Constructs and Transformed
Mammalian Cells; Section (d) Insect Constructs and Transformed
Insect Cells; and Section (e) Bacterial Constructs and Transformed
Bacterial Cells)
[0128] Nucleic acid molecules or fragments thereof of the present
invention are capable of specifically hybridizing to other nucleic
acid molecules under certain circumstances. As used herein, two
nucleic acid molecules are said to be capable of specifically
hybridizing to one another if the two molecules are capable of
forming an anti-parallel, double-stranded nucleic acid structure. A
nucleic acid molecule is said to be the "complement" of another
nucleic acid molecule if they exhibit complete complementarity. As
used herein, molecules are said to exhibit "complete
complementarity" when every nucleotide of one of the molecules is
complementary to a nucleotide of the other. Two molecules are said
to be "minimally complementary" if they can hybridize to one
another with sufficient stability to permit them to remain annealed
to one another under at least conventional "low-stringency"
conditions. Similarly, the molecules are said to be "complementary"
if they can hybridize to one another with sufficient stability to
permit them to remain annealed to one another under conventional
"high-stringency" conditions. Conventional stringency conditions
are described by Sambrook et al., Molecular Cloning, A Laboratory
Manual, 2nd Ed., Cold Spring Harbor Press, Cold Spring Harbor, N.Y.
(1989) and by Haymes et al., Nucleic Acid Hybridization, A
Practical Approach, IRL Press, Washington, D.C. (1985), the
entirety of which is herein incorporated by reference. Departures
from complete complementarity are therefore permissible, as long as
such departures do not completely preclude the capacity of the
molecules to form a double-stranded structure. Thus, in order for a
nucleic acid molecule to serve as a primer or probe it need only be
sufficiently complementary in sequence to be able to form a stable
double-stranded structure under the particular solvent and salt
concentrations employed.
[0129] Appropriate stringency conditions which promote DNA
hybridization, for example, 6.0.times. sodium chloride/sodium
citrate (SSC) at about 45.degree. C., followed by a wash of
2.0.times.SSC at 50.degree. C., are known to those skilled in the
art or can be found in Current Protocols in Molecular Biology, John
Wiley & Sons, N.Y. (1989), 6.3.1-6.3.6. For example, the salt
concentration in the wash step can be selected from a low
stringency of about 2.0.times.SSC at 50.degree. C. to a high
stringency of about 0.2.times.SSC at 50.degree. C. In addition, the
temperature in the wash step can be increased from low stringency
conditions at room temperature, about 22.degree. C., to high
stringency conditions at about 65.degree. C. Both temperature and
salt may be varied, or either the temperature or the salt
concentration may be held constant while the other variable is
changed.
[0130] In a preferred embodiment, a nucleic acid of the present
invention will specifically hybridize to one or more of the nucleic
acid molecules set forth in SEQ ID NO: 1 through SEQ ID NO: 3204 or
complements thereof under moderately stringent conditions, for
example at about 2.0.times.SSC and about 65.degree. C.
[0131] In a particularly preferred embodiment, a nucleic acid of
the present invention will include those nucleic acid molecules
that specifically hybridize to one or more of the nucleic acid
molecules set forth in SEQ ID NO: 1 through SEQ ID NO: 3204 or
complements thereof under high stringency conditions such as
0.2.times.SSC and about 65.degree. C.
[0132] In one aspect of the present invention, the nucleic acid
molecules of the present invention have one or more of the nucleic
acid sequences set forth in SEQ ID NO: 1 through SEQ ID NO: 3204 or
complements thereof. In another aspect of the present invention,
one or more of the nucleic acid molecules of the present invention
share between 100% and 90% sequence identity with one or more of
the nucleic acid sequences set forth in SEQ ID NO: 1 through SEQ ID
NO: 3204 or complements thereof. In a further aspect of the present
invention, one or more of the nucleic acid molecules of the present
invention share between 100% and 95% sequence identity with one or
more of the nucleic acid sequences set forth in SEQ ID NO: 1
through SEQ ID NO: 3204 or complements thereof. In a more preferred
aspect of the present invention, one or more of the nucleic acid
molecules of the present invention share between 100% and 98%
sequence identity with one or more of the nucleic acid sequences
set forth in SEQ ID NO: 1 through SEQ ID NO: 3204 or complements
thereof. In an even more preferred aspect of the present invention,
one or more of the nucleic acid molecules of the present invention
share between 100% and 99% sequence identity with one or more of
the sequences set forth in SEQ ID NO: 1 through SEQ ID NO: 3204 or
complements thereof.
[0133] In a further more preferred aspect of the present invention,
one or more of the nucleic acid molecules of the present invention
exhibit 100% sequence identity with a nucleic acid molecule present
within MONN01, SATMON001 through SATMON031, SATMON033, SATMON034,
SATMON.about.001, SATMONN01, SATMONN04 through SATMONN006, CMz029
through CMz031, CMz033, CMz035 through CMz037, CMz039 through
CMz042, CMz044 through CMz045, CMz047 through CMz050, SOYMON001
through SOYMON038, Soy51 through Soy56, Soy58 through Soy62, Soy65
through Soy66, Soy 68 through Soy73 and Soy76 through Soy77, Lib9,
Lib22 through Lib25, Lib35, Lib80 through Lib81, Lib 144, Lib146,
Lib147, Lib190, Lib3032 through Lib3036 and Lib3099 (Monsanto
Company, St. Loius, Mo. U.S.A.).
[0134] (i) Nucleic Acid Molecules Encoding Proteins or Fragments
Thereof
[0135] Nucleic acid molecules of the present invention can comprise
sequences that encode a methionine pathway protein or fragment
thereof. Such proteins or fragments thereof include homologues of
known proteins in other organisms.
[0136] In a preferred embodiment of the present invention, a maize
or soybean protein homologue or fragment thereof of the present
invention is a homologue of another plant protein. In another
preferred embodiment of the present invention, a maize or soybean
protein homologue or fragment thereof of the present invention is a
homologue of a fungal protein. In another preferred embodiment of
the present invention, a maize or soybean protein homologue of the
present invention is a homologue of mammalian protein. In another
preferred embodiment of the present invention, a maize or soybean
protein homologue or fragment thereof of the present invention is a
homologue of a bacterial protein. In another preferred embodiment
of the present invention, a soybean protein homologue or fragment
thereof of the present invention is a homologue of a maize protein.
In another preferred embodiment of the present invention, a maize
protein homologue or fragment thereof of the present invention is a
homologue of a soybean protein.
[0137] In a preferred embodiment of the present invention, the
nucleic molecule of the present invention encodes a maize or
soybean homologue protein or fragment thereof where a maize or
soybean homologue protein exhibits a BLAST probability score of
greater than 1E-12, preferably a BLAST probability score of between
about 1E-30 and about 1E-12, even more preferably a BLAST
probability score of greater than 1E-30 with its homologue.
[0138] In another preferred embodiment of the present invention,
the nucleic acid molecule encoding a maize or soybean protein
homologue or fragment thereof or fragment thereof exhibits a %
identity with its homologue of between about 25% and about 40%,
more preferably of between about 40 and about 70%, even more
preferably of between about 70% and about 90% and even more
preferably between about 90% and 99%. In another preferred
embodiment, of the present invention, a maize or soybean protein
homologue or fragments thereof exhibits a % identity with its
homologue of 100%.
[0139] In a preferred embodiment of the present invention, the
nucleic molecule of the present invention encodes a maize or
soybean homologue protein or fragment thereof where a maize or
soybean homologue protein exhibits a BLAST score of greater than
120, preferably a BLAST score of between about 1450 and about 120,
even more preferably a BLAST score of greater than 1450 with its
homologue.
[0140] Nucleic acid molecules of the present invention also include
non-maize, non-soybean homologues. Preferred non-homologues are
selected from the group consisting of alfalfa, Arabidopsis, barley,
Brassica, broccoli, cabbage, citrus, cotton, garlic, oat, oilseed
rape, onion, canola, flax, an ornamental plant, pea, peanut,
pepper, potato, rice, rye, sorghum, strawberry, sugarcane,
sugarbeet, tomato, wheat, poplar, pine, fir, eucalyptus, apple,
lettuce, lentils, grape, banana, tea, turf grasses, sunflower, oil
palm and Phaseolus.
[0141] In a preferred embodiment, nucleic acid molecules having SEQ
ID NO: 1 through SEQ ID NO: 3204 or complements and fragments of
either can be utilized to obtain such homologues.
[0142] The degeneracy of the genetic code, which allows different
nucleic acid sequences to code for the same protein or peptide, is
known in the literature. (U.S. Pat. No. 4,757,006, the entirety of
which is herein incorporated by reference).
[0143] In an aspect of the present invention, one or more of the
nucleic acid molecules of the present invention differ in nucleic
acid sequence from those encoding maize or soybean homologue or
fragment thereof in SEQ ID NO: 1 through SEQ ID NO: 3204 due to the
degeneracy in the genetic code in that they encode the same protein
but differ in nucleic acid sequence.
[0144] In another further aspect of the present invention, one or
more of the nucleic acid molecules of the present invention differ
in nucleic acid sequence from those encoding maize or soybean
homologue or fragment thereof in SEQ ID NO: 1 through SEQ ID NO:
3204 due to fact that the different nucleic acid sequence encodes a
protein having one or more conservative amino acid residue.
Examples of conservative substitutions are set forth in Table 1. It
is understood that codons capable of coding for such conservative
substitutions are known in the art.
TABLE-US-00001 TABLE 1 Original Residue Conservative Substitutions
Ala Ser Arg Lys Asn Gln; His Asp Glu Cys Ser; Ala Gln Asn Glu Asp
Gly Pro His Asn; Gln Ile Leu; Val Leu Ile; Val Lys Arg; Gln; Glu
Met Leu; Ile Phe Met; Leu; Tyr Ser Thr Thr Ser Trp Tyr Tyr Trp; Phe
Val Ile; Leu
[0145] In a further aspect of the present invention, one or more of
the nucleic acid molecules of the present invention differ in
nucleic acid sequence from those encoding a maize or soybean
homologue or fragment thereof set forth in SEQ ID NO: 1 through SEQ
ID NO: 3204 or fragment thereof due to the fact that one or more
codons encoding an amino acid has been substituted for a codon that
encodes a nonessential substitution of the amino acid originally
encoded.
[0146] Agents of the present invention include nucleic acid
molecules that encode a maize or soybean methionine pathway protein
or fragment thereof and particularly substantially purified nucleic
acid molecules selected from the group consisting of a nucleic acid
molecule that encodes a maize or soybean methionine
adenosyltransferase protein or fragment thereof, a nucleic acid
molecule that encodes a maize or soybean S-adenosylmethionine
decarboxylase protein or fragment thereof, a nucleic acid molecule
that encodes a maize or soybean aspartate kinase protein or
fragment thereof, a nucleic acid molecule that encode a maize or
soybean aspartate-semialdehyde dehydrogenase protein or fragment
thereof, a nucleic acid molecule that encodes a maize or soybean
O-succinylhomoserine (thiol)-lyase protein or fragment thereof, a
nucleic acid molecule that encodes a maize or soybean cystathionine
.beta.-lyase protein or fragment thereof, a nucleic acid molecule
that encodes a maize or soybean
5-methyltetrahydropteroyltriglutamate-homocysteine-S-methyltransferase
protein or fragment thereof, a nucleic acid molecule that encodes a
maize or soybean adenosylhomocysteine protein or fragment thereof,
a nucleic acid molecule that encodes a maize or soybean
cystathionine .beta.-synthase protein or fragment thereof, a
nucleic acid molecule that encodes a maize or soybean cystathionine
.gamma.-lyase protein or fragment thereof, and a nucleic acid
molecule that encodes a maize or soybean O-acetylhomoserine
(thiol)-lyase protein or fragment thereof.
[0147] Non-limiting examples of such nucleic acid molecules of the
present invention are nucleic acid molecules comprising: SEQ ID NO:
1 through SEQ ID NO: 3204 or fragment thereof that encode for a
methionine pathway protein or fragment thereof, SEQ ID NO: 1
through SEQ ID NO: 429 and SEQ ID NO: 1635 through SEQ ID NO: 2479
or fragment thereof that encode for a methionine
adenosyltransferase protein or fragment thereof, SEQ ID NO: 430
through SEQ ID NO: 857 and SEQ ID NO: 2480 through SEQ ID NO: 2623
or fragment thereof that encode for a S-adenosylmethionine
decarboxylase protein or fragment thereof, SEQ ID NO: 858 through
SEQ ID NO: 900 and SEQ ID NO: 2624 through SEQ ID NO: 2648 or
fragment thereof that encode for a aspartate kinase protein or
fragment thereof, SEQ ID NO: 901 through SEQ ID NO: 904 and SEQ ID
NO: 2649 through SEQ ID NO: 2654 or fragment thereof that encode
for a aspartate-semialdehyde dehydrogenase protein or fragment
thereof, SEQ ID NO: 905 through SEQ ID NO: 953 and SEQ ID NO: 2655
through SEQ ID NO: 2660 or fragment thereof that encode for a
O-succinylhomoserine (thiol)-lyase protein or fragment thereof, SEQ
ID NO: 954 through SEQ ID NO: 963 and SEQ ID NO: 2655 through SEQ
ID NO: 2660 or fragment thereof that encode for a cystathionine
.beta.-lyase protein or fragment thereof, SEQ ID NO: 964 through
SEQ ID NO: 1353 and SEQ ID NO: 2666 through SEQ ID NO: 2992 or
fragment thereof that encode for a
5-methyltetrahydropteroyltriglutamate-homocysteine-S-methyltransfer-
ase protein or fragment thereof, SEQ ID NO: 1354 through SEQ ID NO:
1630 and SEQ ID NO: 2993 through SEQ ID NO: 3199 or fragment
thereof that encode for an adenosylhomocysteinase protein or
fragment thereof, SEQ ID NO: 1631 through SEQ ID NO: 1632 or
fragment thereof that encode for a cystathionine .beta.-synthase
protein or fragment thereof, SEQ ID NO: 1633 through SEQ ID NO:
1634 and SEQ ID NO: 3203 through SEQ ID NO: 3204 or fragment
thereof that encode for a cystathionine .gamma.-lyase protein or
fragment thereof, and SEQ ID NO: 3200 through SEQ ID NO: 3202 or
fragment thereof that encode for an O-acetylhomoserine
(thiol)-lyase protein or fragment thereof.
[0148] A nucleic acid molecule of the present invention can also
encode an homologue of a maize or soybean methionine
adenosyltransferase or fragment thereof, a maize or soybean
S-adenosylmethionine decarboxylase or fragment thereof, a maize or
soybean aspartate kinase or fragment thereof, a maize or soybean
aspartate-semialdehyde dehydrogenase or fragment thereof, a maize
or soybean O-succinylhomoserine (thiol)-lyase or fragment thereof,
a maize or soybean cystathionine .beta.-lyase or fragment thereof,
a maize or soybean
5-methyltetrahydropteroyltriglutamate-homocysteine-S-methyltransferase
or fragment thereof, a maize or soybean adenosylhomocysteinease or
fragment thereof, a maize or soybean cystathionine .beta.-synthase
or fragment thereof, a maize or soybean cystathionine .gamma.-lyase
or fragment thereof or a maize or soybean O-acetylhomoserine
(thiol)-lyase or fragment thereof. As used herein a homologue
protein molecule or fragment thereof is a counterpart protein
molecule or fragment thereof in a second species (e.g., maize
methionine adenosyltransferase protein is a homologue of
Arabidopsis' methionine adenosyltransferase protein).
[0149] (ii) Nucleic Acid Molecule Markers and Probes
[0150] One aspect of the present invention concerns markers that
include nucleic acid molecules SEQ ID NO: 1 through SEQ ID NO: 3204
or complements thereof or fragments of either that can act as
markers. Genetic markers of the present invention include
"dominant" or "codominant" markers. "Codominant markers" reveal the
presence of two or more alleles (two per diploid individual) at a
locus. "Dominant markers" reveal the presence of only a single
allele per locus. The presence of the dominant marker phenotype
(e.g., a band of DNA) is an indication that one allele is present
in either the homozygous or heterozygous condition. The absence of
the dominant marker phenotype (e.g. absence of a DNA band) is
merely evidence that "some other" undefined allele is present. In
the case of populations where individuals are predominantly
homozygous and loci are predominately dimorphic, dominant and
codominant markers can be equally valuable. As populations become
more heterozygous and multi-allelic, codominant markers often
become more informative of the genotype than dominant markers.
Marker molecules can be, for example, capable of detecting
polymorphisms such as single nucleotide polymorphisms (SNPs).
[0151] SNPs are single base changes in genomic DNA sequence. They
occur at greater frequency and are spaced with a greater uniformly
throughout a genome than other reported forms of polymorphism. The
greater frequency and uniformity of SNPs means that there is
greater probability that such a polymorphism will be found near or
in a genetic locus of interest than would be the case for other
polymorphisms. SNPs are located in protein-coding regions and
noncoding regions of a genome. Some of these SNPs may result in
defective or variant protein expression (e.g., as a results of
mutations or defective splicing). Analysis (genotyping) of
characterized SNPs can require only a plus/minus assay rather than
a lengthy measurement, permitting easier automation.
[0152] SNPs can be characterized using any of a variety of methods.
Such methods include the direct or indirect sequencing of the site,
the use of restriction enzymes (Botstein et al., Am. J. Hum. Genet.
32:314-331 (1980), the entirety of which is herein incorporated
reference; Konieczny and Ausubel, Plant J. 4:403-410 (1993), the
entirety of which is herein incorporated by reference), enzymatic
and chemical mismatch assays (Myers et al., Nature 313:495-498
(1985), the entirety of which is herein incorporated by reference),
allele-specific PCR (Newton et al., Nucl. Acids Res. 17:2503-2516
(1989), the entirety of which is herein incorporated by reference;
Wu et al., Proc. Natl. Acad. Sci. (U.S.A.) 86:2757-2760 (1989), the
entirety of which is herein incorporated by reference), ligase
chain reaction (Barany, Proc. Natl. Acad. Sci. (U.S.A.) 88:189-193
(1991), the entirety of which is herein incorporated by reference),
single-strand conformation polymorphism analysis (Labrune et al.,
Am. J. Hum. Genet. 48: 1115-1120 (1991), the entirety of which is
herein incorporated by reference), primer-directed nucleotide
incorporation assays (Kuppuswami et al., Proc. Natl. Acad. Sci. USA
88:1143-1147 (1991), the entirety of which is herein incorporated
by reference), dideoxy fingerprinting (Sarkar et al., Genomics
13:441-443 (1992), the entirety of which is herein incorporated by
reference), solid-phase ELISA-based oligonucleotide ligation assays
(Nikiforov et al., Nucl. Acids Res. 22:4167-4175 (1994), the
entirety of which is herein incorporated by reference),
oligonucleotide fluorescence-quenching assays (Livak et al., PCR
Methods Appl. 4:357-362 (1995), the entirety of which is herein
incorporated by reference), 5'-nuclease allele-specific
hybridization TaqMan assay (Livak et al., Nature Genet. 9:341-342
(1995), the entirety of which is herein incorporated by reference),
template-directed dye-terminator incorporation (TDI) assay (Chen
and Kwok, Nucl. Acids Res. 25:347-353 (1997), the entirety of which
is herein incorporated by reference), allele-specific molecular
beacon assay (Tyagi et al., Nature Biotech. 16: 49-53 (1998), the
entirety of which is herein incorporated by reference), PinPoint
assay (Haff and Smirnov, Genome Res. 7: 378-388 (1997), the
entirety of which is herein incorporated by reference) and dCAPS
analysis (Neff et al., Plant 114:387-392 (1998), the entirety of
which is herein incorporated by reference).
[0153] Additional markers, such as AFLP markers, RFLP markers and
RAPD markers, can be utilized (Walton, Seed World 22-29 (July,
1993), the entirety of which is herein incorporated by reference;
Burow and Blake, Molecular Dissection of Complex Traits, 13-29,
Paterson (ed.), CRC Press, New York (1988), the entirety of which
is herein incorporated by reference). DNA markers can be developed
from nucleic acid molecules using restriction endonucleases, the
PCR and/or DNA sequence information. RFLP markers result from
single base changes or insertions/deletions. These codominant
markers are highly abundant in plant genomes, have a medium level
of polymorphism and are developed by a combination of restriction
endonuclease digestion and Southern blotting hybridization. CAPS
are similarly developed from restriction nuclease digestion but
only of specific PCR products. These markers are also codominant,
have a medium level of polymorphism and are highly abundant in the
genome. The CAPS result from single base changes and
insertions/deletions.
[0154] Another marker type, RAPDs, are developed from DNA
amplification with random primers and result from single base
changes and insertions/deletions in plant genomes. They are
dominant markers with a medium level of polymorphisms and are
highly abundant. AFLP markers require using the PCR on a subset of
restriction fragments from extended adapter primers. These markers
are both dominant and codominant are highly abundant in genomes and
exhibit a medium level of polymorphism.
[0155] SSRs require DNA sequence information. These codominant
markers result from repeat length changes, are highly polymorphic
and do not exhibit as high a degree of abundance in the genome as
CAPS, AFLPs and RAPDs SNPs also require DNA sequence information.
These codominant markers result from single base substitutions.
They are highly abundant and exhibit a medium of polymorphism
(Rafalski et al., In: Nonmammalian Genomic Analysis, Birren and Lai
(ed.), Academic Press, San Diego, Calif., pp. 75-134 (1996), the
entirety of which is herein incorporated by reference). It is
understood that a nucleic acid molecule of the present invention
may be used as a marker.
[0156] A PCR probe is a nucleic acid molecule capable of initiating
a polymerase activity while in a double-stranded structure to with
another nucleic acid. Various methods for determining the structure
of PCR probes and PCR techniques exist in the art. Computer
generated searches using programs such as Primer3
(www-genome.wi.mit.edu/cgi-bin/primer/primer3.cgi), STSPipeline
(www-genome.wi.mit.edu/cgi-bin/www-STS_Pipeline), or GeneUp (Pesole
et al., BioTechniques 25:112-123 (1998) the entirety of which is
herein incorporated by reference), for example, can be used to
identify potential PCR primers.
[0157] It is understood that a fragment of one or more of the
nucleic acid molecules of the present invention may be a probe and
specifically a PCR probe.
[0158] (b) Protein and Peptide Molecules
[0159] A class of agents comprises one or more of the protein or
fragments thereof or peptide molecules encoded by SEQ ID NO: 1
through SEQ ID NO: 3204 or one or more of the protein or fragment
thereof and peptide molecules encoded by other nucleic acid agents
of the present invention. As used herein, the term "protein
molecule" or "peptide molecule" includes any molecule that
comprises five or more amino acids. It is well known in the art
that proteins may undergo modification, including
post-translational modifications, such as, but not limited to,
disulfide bond formation, glycosylation, phosphorylation, or
oligomerization. Thus, as used herein, the term "protein molecule"
or "peptide molecule" includes any protein molecule that is
modified by any biological or non-biological process. The terms
"amino acid" and "amino acids" refer to all naturally occurring
L-amino acids. This definition is meant to include norleucine,
ornithine, homocysteine and homoserine.
[0160] Non-limiting examples of the protein or fragment thereof of
the present invention include a maize or soybean methionine pathway
protein or fragment thereof; a maize or soybean methionine
adenosyltransferase or fragment thereof, a maize or soybean
S-adenosylmethionine decarboxylase or fragment thereof, a maize or
soybean aspartate kinase or fragment thereof, a maize or soybean
aspartate-semialdehyde dehydrogenase or fragment thereof, a maize
or soybean O-succinylhomoserine (thiol)-lyase or fragment thereof,
a maize or soybean cystathionine .beta.-lyase or fragment thereof,
a maize or soybean
5-methyltetrahydropteroyltriglutamate-homocysteine-S-methyltransferase
or fragment thereof, a maize or soybean adenosylhomocysteinase or
fragment thereof, a maize or soybean cystathionine .beta.-synthase
or fragment thereof, a maize or soybean cystathionine .gamma.-lyase
or fragment thereof or a maize or soybean O-acetylhomoserine
(thiol)-lyase or fragment thereof.
[0161] Non-limiting examples of the protein or fragment molecules
of the present invention are an methionine pathway protein or
fragment thereof encoded by: SEQ ID NO: 1 through SEQ ID NO: 3204
or fragment thereof that encode for a methionine pathway protein or
fragment thereof, SEQ ID NO: 1 through SEQ ID NO: 429 and SEQ ID
NO: 1635 through SEQ ID NO: 2479 or fragment thereof that encode
for a methionine adenosyltransferase protein or fragment thereof,
SEQ ID NO: 430 through SEQ ID NO: 857 and SEQ ID NO: 2480 through
SEQ ID NO: 2623 or fragment thereof that encode for a
S-adenosylmethionine decarboxylase protein or fragment thereof, SEQ
ID NO: 858 through SEQ ID NO: 900 and SEQ ID NO: 2624 through SEQ
ID NO: 2648 or fragment thereof that encode for a aspartate kinase
protein or fragment thereof, SEQ ID NO: 901 through SEQ ID NO: 904
and SEQ ID NO: 2649 through SEQ ID NO: 2654 or fragment thereof
that encode for a aspartate-semialdehyde dehydrogenase protein or
fragment thereof, SEQ ID NO: 905 through SEQ ID NO: 953 and SEQ ID
NO: 2655 through SEQ ID NO: 2660 or fragment thereof that encode
for a O-succinylhomoserine (thiol)-lyase protein or fragment
thereof, SEQ ID NO: 954 through SEQ ID NO: 963 and SEQ ID NO: 2655
through SEQ ID NO: 2660 or fragment thereof that encode for a
cystathionine .beta.-lyase or fragment thereof, SEQ ID NO: 964
through SEQ ID NO: 1353 and SEQ ID NO: 2666 through SEQ ID NO: 2992
or fragment thereof that encode for a
5-methyltetrahydropteroyltriglutamate-homocysteine-S-methyltransferase
protein or fragment thereof, SEQ ID NO: 1354 through SEQ ID NO:
1630 and SEQ ID NO: 2993 through SEQ ID NO: 3199 or fragment
thereof that encode for an adenosylhomocysteinase protein or
fragment thereof, SEQ ID NO: 1631 through SEQ ID NO: 1632 or
fragment thereof that encode for a cystathionine fl-synthase
protein or fragment thereof, SEQ ID NO: 1633 through SEQ ID NO:
1634 and SEQ ID NO: 3203 through SEQ ID NO: 3204 or fragment
thereof that encode for a cystathionine .gamma.-lyase protein or
fragment thereof, and SEQ ID NO: 3200 through SEQ ID NO: 3202 or
fragment thereof that encode for an O-acetylhomoserine
(thiol)-lyase protein or fragment thereof.
[0162] One or more of the protein or fragment of peptide molecules
may be produced via chemical synthesis, or more preferably, by
expressing in a suitable bacterial or eucaryotic host. Suitable
methods for expression are described by Sambrook et al., (In:
Molecular Cloning, A Laboratory Manual, 2nd Edition, Cold Spring
Harbor Press, Cold Spring Harbor, N.Y. (1989)), or similar texts.
For example, the protein may be expressed in, for example, Uses of
the Agents of the Invention, Section (a) Plant Constructs and Plant
Transformants; Section (b) Fungal Constructs and Fungal
Transformants; Section (c) Mammalian Constructs and Transformed
Mammalian Cells; Section (d) Insect Constructs and Transformed
Insect Cells; and Section (e) Bacterial Constructs and Transformed
Bacterial Cells.
[0163] A "protein fragment" is a peptide or polypeptide molecule
whose amino acid sequence comprises a subset of the amino acid
sequence of that protein. A protein or fragment thereof that
comprises one or more additional peptide regions not derived from
that protein' is a "fusion" protein. Such molecules may be
derivatized to contain carbohydrate or other moieties (such as
keyhole limpet hemocyanin, etc.). Fusion protein or peptide
molecules of the present invention are preferably produced via
recombinant means.
[0164] Another class of agents comprise protein or peptide
molecules or fragments or fusions thereof encoded by SEQ ID NO: 1
through SEQ ID NO: 3204 or complements thereof in which
conservative, non-essential or non-relevant amino acid residues
have been added, replaced or deleted. Computerized means for
designing modifications in protein structure are known in the art
(Dahiyat and Mayo, Science 278:82-87 (1997), the entirety of which
is herein incorporated by reference).
[0165] The protein molecules of the present invention include plant
homologue proteins. An example of such a homologue is a homologue
protein of a non-maize or non soybean plant species, that include
but not limited to alfalfa, Arabidopsis, barley, Brassica,
broccoli, cabbage, citrus, cotton, garlic, oat, oilseed rape,
onion, canola, flax, an ornamental plant, pea, peanut, pepper,
potato, rice, rye, sorghum, strawberry, sugarcane, sugarbeet,
tomato, wheat, poplar, pine, fir, eucalyptus, apple, lettuce,
lentils, grape, banana, tea, turf grasses, sunflower, oil palm,
Phaseolus etc. Particularly preferred non-maize or non-soybean for
use for the isolation of homologs would include, Arabidopsis,
barley, cotton, oat, oilseed rape, rice, canola, ornamentals,
sugarcane, sugarbeet, tomato, potato, wheat and turf grasses. Such
a homologue can be obtained by any of a variety of methods. Most
preferably, as indicated above, one or more of the disclosed
sequences (SEQ ID NO: 1 through SEQ ID NO: 3204 or complements
thereof) will be used to define a pair of primers that may be used
to isolate the homologue-encoding nucleic acid molecules from any
desired species. Such molecules can be expressed to yield
homologues by recombinant means.
[0166] (c) Antibodies
[0167] One aspect of the present invention concerns antibodies,
single-chain antigen binding molecules, or other proteins that
specifically bind to one or more of the protein or peptide
molecules of the present invention and their homologues, fusions or
fragments. Such antibodies may be used to quantitatively or
qualitatively detect the protein or peptide molecules of the
present invention. As used herein, an antibody or peptide is said
to "specifically bind" to a protein or peptide molecule of the
present invention if such binding is not competitively inhibited by
the presence of non-related molecules.
[0168] Nucleic acid molecules that encode all or part of the
protein of the present invention can be expressed, via recombinant
means, to yield protein or peptides that can in turn be used to
elicit antibodies that are capable of binding the expressed protein
or peptide. Such antibodies may be used in immunoassays for that
protein. Such protein-encoding molecules, or their fragments may be
a "fusion" molecule (i.e., a part of a larger nucleic acid
molecule) such that, upon expression, a fusion protein is produced.
It is understood that any of the nucleic acid molecules of the
present invention may be expressed, via recombinant means, to yield
proteins or peptides encoded by these nucleic acid molecules.
[0169] The antibodies that specifically bind proteins and protein
fragments of the present invention may be polyclonal or monoclonal
and may comprise intact immunoglobulins, or antigen binding
portions of immunoglobulins fragments (such as (F(ab'),
F(ab').sub.2), or single-chain immunoglobulins producible, for
example, via recombinant means. It is understood that practitioners
are familiar with the standard resource materials which describe
specific conditions and procedures for the construction,
manipulation and isolation of antibodies (see, for example, Harlow
and Lane, In: Antibodies: A Laboratory Manual, Cold Spring Harbor
Press, Cold Spring Harbor, N.Y. (1988), the entirety of which is
herein incorporated by reference).
[0170] Murine monoclonal antibodies are particularly preferred.
BALB/c mice are preferred for this purpose, however, equivalent
strains may also be used. The animals are preferably immunized with
approximately 25 .mu.g of purified protein (or fragment thereof)
that has been emulsified in a suitable adjuvant (such as TiterMax
adjuvant (Vaxcel, Norcross, Ga.)). Immunization is preferably
conducted at two intramuscular sites, one intraperitoneal site and
one subcutaneous site at the base of the tail. An additional i.v.
injection of approximately 25 .mu.g of antigen is preferably given
in normal saline three weeks later. After approximately 11 days
following the second injection, the mice may be bled and the blood
screened for the presence of anti-protein or peptide antibodies.
Preferably, a direct binding Enzyme-Linked Immunoassay (ELISA) is
employed for this purpose.
[0171] More preferably, the mouse having the highest antibody titer
is given a third i.v. injection of approximately 25 .mu.g of the
same protein or fragment. The splenic leukocytes from this animal
may be recovered 3 days later and then permitted to fuse, most
preferably, using polyethylene glycol, with cells of a suitable
myeloma cell line (such as, for example, the P3X63Ag8.653 myeloma
cell line). Hybridoma cells are selected by culturing the cells
under "HAT" (hypoxanthine-aminopterin-thymine) selection for about
one week. The resulting clones may then be screened for their
capacity to produce monoclonal antibodies ("mAbs"), preferably by
direct ELISA.
[0172] In one embodiment, anti-protein or peptide monoclonal
antibodies are isolated using a fusion of a protein or peptide of
the present invention, or conjugate of a protein or peptide of the
present invention, as immunogens. Thus, for example, a group of
mice can be immunized using a fusion protein emulsified in Freund's
complete adjuvant (e.g. approximately 50 .mu.g of antigen per
immunization). At three week intervals, an identical amount of
antigen is emulsified in Freund's incomplete adjuvant and used to
immunize the animals. Ten days following the third immunization,
serum samples are taken and evaluated for the presence of antibody.
If antibody titers are too low, a fourth booster can be employed.
Polysera capable of binding the protein or peptide can also be
obtained using this method.
[0173] In a preferred procedure for obtaining monoclonal
antibodies, the spleens of the above-described immunized mice are
removed, disrupted and immune splenocytes are isolated over a
ficoll gradient. The isolated splenocytes are fused, using
polyethylene glycol with BALB/c-derived HGPRT (hypoxanthine guanine
phosphoribosyl transferase) deficient P3x63xAg8.653 plasmacytoma
cells. The fused cells are plated into 96 well microtiter plates
and screened for hybridoma fusion cells by their capacity to grow
in culture medium supplemented with hypothanthine, aminopterin and
thymidine for approximately 2-3 weeks.
[0174] Hybridoma cells that arise from such incubation are
preferably screened for their capacity to produce an immunoglobulin
that binds to a protein of interest. An indirect ELISA may be used
for this purpose. In brief, the supernatants of hybridomas are
incubated in microtiter wells that contain immobilized protein.
After washing, the titer of bound immunoglobulin can be determined
using, for example, a goat anti-mouse antibody conjugated to
horseradish peroxidase. After additional washing, the amount of
immobilized enzyme is determined (for example through the use of a
chromogenic substrate). Such screening is performed as quickly as
possible after the identification of the hybridoma in order to
ensure that a desired clone is not overgrown by non-secreting
neighbor cells. Desirably, the fusion plates are screened several
times since the rates of hybridoma growth vary. In a preferred
sub-embodiment, a different antigenic form may be used to screen
the hybridoma. Thus, for example, the splenocytes may be immunized
with one immunogen, but the resulting hybridomas can be screened
using a different immunogen. It is understood that any of the
protein or peptide molecules of the present invention may be used
to raise antibodies.
[0175] As discussed below, such antibody molecules or their
fragments may be used for diagnostic purposes. Where the antibodies
are intended for diagnostic purposes, it may be desirable to
derivatize them, for example with a ligand group (such as biotin)
or a detectable marker group (such as a fluorescent group, a
radioisotope or an enzyme).
[0176] The ability to produce antibodies that bind the protein or
peptide molecules of the present invention permits the
identification of mimetic compounds of those moleculeS. A "mimetic
compound" is a compound that is not that compound, or a fragment of
that compound, but which nonetheless exhibits an ability to
specifically bind to antibodies directed against that compound.
[0177] It is understood that any of the agents of the present
invention can be substantially purified and/or be biologically
active and/or recombinant.
Uses of the Agents of the Invention
[0178] Nucleic acid molecules and fragments thereof of the present
invention may be employed to obtain other nucleic acid molecules
from the same species (e.g., ESTs or fragment thereof from maize
may be utilized to obtain other nucleic acid molecules from maize).
Such nucleic acid molecules include the nucleic acid molecules that
encode the complete coding sequence of a protein and promoters and
flanking sequences of such molecules. In addition, such nucleic
acid molecules include nucleic acid molecules that encode for other
isozymes or gene family members. Such molecules can be readily
obtained by using the above-described nucleic acid molecules or
fragments thereof to screen cDNA or genomic libraries obtained from
maize or soybean. Methods for forming such libraries are well known
in the art.
[0179] Nucleic acid molecules and fragments thereof of the present
invention may also be employed to obtain nucleic acid homologues.
Such homologues include the nucleic acid molecule of other plants
or other organisms (e.g., alfalfa, Arabidopsis, barley, Brassica,
broccoli, cabbage, citrus, cotton, garlic, oat, oilseed rape,
onion, canola, flax, an ornamental plant, pea, peanut, pepper,
potato, rice, rye, sorghum, strawberry, sugarcane, sugarbeet,
tomato, wheat, poplar, pine, fir, eucalyptus, apple, lettuce,
lentils, grape, banana, tea, turf grasses, sunflower, oil palm,
Phaseolus, etc.) including the nucleic acid molecules that encode,
in whole or in part, protein homologues of other plant species or
other organisms, sequences of genetic elements such as promoters
and transcriptional regulatory elements. Such molecules can be
readily obtained by using the above-described nucleic acid
molecules or fragments thereof to screen cDNA or genomic libraries
obtained from such plant species. Methods for forming such
libraries are well known in the art. Such homologue molecules may
differ in their nucleotide sequences from those found in one or
more of SEQ ID NO: 1 through SEQ ID NO: 3204 or complements thereof
because complete complementarity is not needed for stable
hybridization. The nucleic acid molecules of the present invention
therefore also include molecules that, although capable of
specifically hybridizing with the nucleic acid molecules may lack
"complete complementarity."
[0180] Any of a variety of methods may be used to obtain one or
more of the above-described nucleic acid molecules (Zamechik et al,
Proc. Natl. Acad. Sci. (U.S.A.) 83:4143-4146 (1986), the entirety
of which is herein incorporated by reference; Goodchild et al.,
Proc. Natl. Acad. Sci. (U.S.A.) 85:5507-5511 (1988), the entirety
of which is herein incorporated by reference; Wickstrom et al.,
Proc. Natl. Acad. Sci. (U.S.A.) 85:1028-1032 (1988), the entirety
of which is herein incorporated by reference; Holt et al., Molec.
Cell. Biol. 8:963-973 (1988), the entirety of which is herein
incorporated by reference; Gerwirtz et al., Science 242:1303-1306
(1988), the entirety of which is herein incorporated by reference;
Anfossi et al., Proc. Natl. Acad. Sci. (U.S.A.) 86:3379-3383
(1989), the entirety of which is herein incorporated by reference;
Becker et al., EMBO J. 8:3685-3691 (1989); the entirety of which is
herein incorporated by reference). Automated nucleic acid
synthesizers may be employed for this purpose. In lieu of such
synthesis, the disclosed nucleic acid molecules may be used to
define a pair of primers that can be used with the polymerase chain
reaction (Mullis et al., Cold Spring Harbor Symp. Quant. Biol.
51:263-273 (1986); Erlich et al., European Patent 50,424; European
Patent 84,796; European Patent 258,017; European Patent 237,362;
Mullis, European Patent 201,184; Mullis et al., U.S. Pat. No.
4,683,202; Erlich, U.S. Pat. No. 4,582,788; and Saiki et al., U.S.
Pat. No. 4,683,194, all of which are herein incorporated by
reference in their entirety) to amplify and obtain any desired
nucleic acid molecule or fragment.
[0181] Promoter sequence(s) and other genetic elements, including
but not limited to transcriptional regulatory flanking sequences,
associated with one or more of the disclosed nucleic acid sequences
can also be obtained using the disclosed nucleic acid sequence
provided herein. In one embodiment, such sequences are obtained by
incubating EST nucleic acid molecules or preferably fragments
thereof with members of genomic libraries (e.g. maize and soybean)
and recovering clones that hybridize to the EST nucleic acid
molecule or fragment thereof. In a second embodiment, methods of
"chromosome walking," or inverse PCR may be used to obtain such
sequences (Frohman et al., Proc. Natl. Acad. Sci. (U.S.A.)
85:8998-9002 (1988); Ohara et al., Proc. Natl. Acad. Sci. (U.S.A.)
86:5673-5677 (1989); Pang et al., Biotechniques 22:1046-1048
(1977); Huang et al., Methods Mol. Biol. 69:89-96 (1997); Huang et
al., Method Mol. Biol. 67:287-294 (1997); Benkel et al, Genet.
Anal. 13:123-127 (1996); Hartl et al., Methods Mol. Biol.
58:293-301 (1996), all of which are herein incorporated by
reference in their entirety).
[0182] The nucleic acid molecules of the present invention may be
used to isolate promoters of cell enhanced, cell specific, tissue
enhanced, tissue specific, developmentally or environmentally
regulated expression profiles. Isolation and functional analysis of
the 5' flanking promoter sequences of these genes from genomic
libraries, for example, using genomic screening methods and PCR
techniques would result in the isolation of useful promoters and
transcriptional regulatory elements. These methods are known to
those of skill in the art and have been described (See, for
example, Birren et al., Genome Analysis: Analyzing DNA, 1, (1997),
Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y., the
entirety of which is herein incorporated by reference). Promoters
obtained utilizing the nucleic acid molecules of the present
invention could also be modified to affect their control
characteristics. Examples of such modifications would include but
are not limited to enhanced sequences as reported in Uses of the
Agents of the Invention, Section (a) Plant Constructs and Plant
Transformants. Such genetic elements could be used to enhance gene
expression of new and existing traits for cro.sub.p
improvements.
[0183] In one sub-aspect, such an analysis is conducted by
determining the presence and/or identity of polymorphism(s) by one
or more of the nucleic acid molecules of the present invention and
more preferably one or more of the EST nucleic acid molecule or
fragment thereof which are associated with a phenotype, or a
predisposition to that phenotype.
[0184] Any of a variety of molecules can be used to identify such
polymorphism(s). In one embodiment, one or more of the EST nucleic
acid molecules (or a sub-fragment thereof) may be employed as a
marker nucleic acid molecule to identify such polymorphism(s).
Alternatively, such polymorphisms can be detected through the use
of a marker nucleic acid molecule or a marker protein that is
genetically linked to (i.e., a polynucleotide that co-segregates
with) such polymorphism(s).
[0185] In an alternative embodiment, such polymorphisms can be
detected through the use of a marker nucleic acid molecule that is
physically linked to such polymorphism(s). For this purpose, marker
nucleic acid molecules comprising a nucleotide sequence of a
polynucleotide located within 1 mb of the polymorphism(s) and more
preferably within 100 kb of the polymorphism(s) and most preferably
within 10 kb of the polymorphism(s) can be employed.
[0186] The genomes of animals and plants naturally undergo
spontaneous mutation in the course of their continuing evolution
(Gusella, Ann. Rev. Biochem. 55:831-454 (1986)). A "polymorphism"
is a variation or difference in the sequence of the gene or its
flanking regions that arises in some of the members of a species.
The variant sequence and the "original" sequence co-exist in the
species' population. In some instances, such co-existence is in
stable or quasi-stable equilibrium.
[0187] A polymorphism is thus said to be "allelic," in that, due to
the existence of the polymorphism, some members of a species may
have the original sequence (i.e., the original "allele") whereas
other members may have the variant sequence (i.e., the variant
"allele"). In the simplest case, only one variant sequence may
exist and the polymorphism is thus said to be di-allelic. In other
cases, the species' population may contain multiple alleles and the
polymorphism is termed tri-allelic, etc. A single gene may have
multiple different unrelated polymorphisms. For example, it may
have a di-allelic polymorphism at one site and a multi-allelic
polymorphism at another site.
[0188] The variation that defines the polymorphism may range from a
single nucleotide variation to the insertion or deletion of
extended regions within a gene. In some cases, the DNA sequence
variations are in regions of the genome that are characterized by
short tandem repeats (STRs) that include tandem di- or
tri-nucleotide repeated motifs of nucleotides. Polymorphisms
characterized by such tandem repeats are referred to as "variable
number tandem repeat" ("VNTR") polymorphisms. VNTRs have been used
in identity analysis (Weber, U.S. Pat. No. 5,075,217; Armour et
al., FEBS Lett. 307:113-115 (1992); Jones et al., Eur. J. Haematol.
39:144-147 (1987); Horn et al., PCT Patent Application WO91/14003;
Jeffreys, European Patent Application 370,719; Jeffreys, U.S. Pat.
No. 5,175,082; Jeffreys et al, Amer. J. Hum. Genet. 39:11-24
(1986); Jeffreys et al., Nature 316:76-79 (1985); Gray et al.,
Proc. R. Acad. Soc. Lond. 243:241-253 (1991); Moore et al.,
Genomics 10:654-660 (1991); Jeffreys et al., Anim. Genet. 18:1-15
(1987); Hillel et al., Anim. Genet. 20:145-155 (1989); Hillel et
al., Genet. 124:783-789 (1990), all of which are herein
incorporated by reference in their entirety).
[0189] The detection of polymorphic sites in a sample of DNA may be
facilitated through the use of nucleic acid amplification methods.
Such methods specifically increase the concentration of
polynucleotides that span the polymorphic site, or include that
site and sequences located either distal or proximal to it. Such
amplified molecules can be readily detected by gel electrophoresis
or other means.
[0190] The most preferred method of achieving such amplification
employs the polymerase chain reaction ("PCR") (Mullis et al., Cold
Spring Harbor Symp. Quant. Biol. 51:263-273 (1986); Erlich et al.,
European Patent Appln. 50,424; European Patent Appln. 84,796;
European Patent Application 258,017; European Patent Appln.
237,362; Mullis, European Patent Appln. 201,184; Mullis et al.,
U.S. Pat. No. 4,683,202; Erlich, U.S. Pat. No. 4,582,788; and Saiki
et al., U.S. Pat. No. 4,683,194), using primer pairs that are
capable of hybridizing to the proximal sequences that define a
polymorphism in its double-stranded form.
[0191] In lieu of PCR, alternative methods, such as the "Ligase
Chain Reaction" ("LCR") may be used (Barany, Proc. Natl. Acad. Sci.
(U.S.A.) 88:189-193 (1991), the entirety of which is herein
incorporated by reference). LCR uses two pairs of oligonucleotide
probes to exponentially amplify a specific target. The sequences of
each pair of oligonucleotides is selected to permit the pair to
hybridize to abutting sequences of the same strand of the target.
Such hybridization forms a substrate for a template-dependent
ligase. As with PCR, the resulting products thus serve as a
template in subsequent cycles and an exponential amplification of
the desired sequence is obtained.
[0192] LCR can be performed with oligonucleotides having the
proximal and distal sequences of the same strand of a polymorphic
site. In one embodiment, either oligonucleotide will be designed to
include the actual polymorphic site of the polymorphism. In such an
embodiment, the reaction conditions are selected such that the
oligonucleotides can be ligated together only if the target
molecule either contains or lacks the specific nucleotide that is
complementary to the polymorphic site present on the
oligonucleotide. Alternatively, the oligonucleotides may be
selected such that they do not include the polymorphic site (see,
Segev, PCT Application WO 90/01069, the entirety of which is herein
incorporated by reference).
[0193] The "Oligonucleotide Ligation Assay" ("OLA") may
alternatively be employed (Landegren et al., Science 241:1077-1080
(1988), the entirety of which is herein incorporated by reference).
The OLA protocol uses two oligonucleotides which are designed to be
capable of hybridizing to abutting sequences of a single strand of
a target. OLA, like LCR, is particularly suited for the detection
of point mutations. Unlike LCR, however, OLA results in "linear"
rather than exponential amplification of the target sequence.
[0194] Nickerson et al., have described a nucleic acid detection
assay that combines attributes of PCR and OLA (Nickerson et al.,
Proc. Natl. Acad. Sci. (U.S.A.) 87:8923-8927 (1990), the entirety
of which is herein incorporated by reference). In this method, PCR
is used to achieve the exponential amplification of target DNA,
which is then detected using OLA. In addition to requiring multiple
and separate, processing steps, one problem associated with such
combinations is that they inherit all of the problems associated
with PCR and OLA.
[0195] Schemes based on ligation of two (or more) oligonucleotides
in the presence of nucleic acid having the sequence of the
resulting "di-oligonucleotide", thereby amplifying the
di-oligonucleotide, are also known (Wu et al., Genomics 4:560-569
(1989), the entirety of which is herein incorporated by reference)
and may be readily adapted to the purposes of the present
invention.
[0196] Other known nucleic acid amplification procedures, such as
allele-specific oligomers, branched DNA technology,
transcription-based amplification systems, or isothermal
amplification methods may also be used to amplify and analyze such
polymorphisms (Malek et al., U.S. Pat. No. 5,130,238; Davey et al.,
European Patent Application 329,822; Schuster et al., U.S. Pat. No.
5,169,766; Miller et al., PCT Patent Application WO 89/06700; Kwoh
et al., Proc. Natl. Acad. Sci. (U.S.A.) 86:1173-1177 (1989);
Gingeras et al., PCT Patent Application WO 88/10315; Walker et al,
Proc. Natl. Acad. Sci. (U.S.A.) 89:392-396 (1992), all of which are
herein incorporated by reference in their entirety).
[0197] The identification of a polymorphism can be determined in a
variety of ways. By correlating the presence or absence of it in a
plant with the presence or absence of a phenotype, it is possible
to predict the phenotype of that plant. If a polymorphism creates
or destroys a restriction endonuclease cleavage site, or if it
results in the loss or insertion of DNA (e.g., a VNTR
polymorphism), it will alter the size or profile of the DNA
fragments that are generated by digestion with that restriction
endonuclease. As such, individuals that possess a variant sequence
can be distinguished from those having the original sequence by
restriction fragment analysis. Polymorphisms that can be identified
in this manner are termed "restriction fragment length
polymorphisms" ("RFLPs"). RFLPs have been widely used in human and
plant genetic analyses (Glassberg, UK Patent Application 2135774;
Skolnick et al., Cytogen. Cell Genet. 32:58-67 (1982); Botstein et
al., Ann. J. Hum. Genet. 32:314-331 (1980); Fischer et al., (PCT
Application WO90/13668); Uhlen, PCT Application WO90/11369).
[0198] Polymorphisms can also be identified by Single Strand
Conformation Polymorphism (SSCP) analysis. SSCP is a method capable
of identifying most sequence variations in a single strand of DNA,
typically between 150 and 250 nucleotides in length (Elles, Methods
in Molecular Medicine Molecular Diagnosis of Genetic Diseases,
Humana Press (1996), the entirety of which is herein incorporated
by reference); Orita et al., Genomics 5:874-879 (1989), the
entirety of which is herein incorporated by reference). Under
denaturing conditions a single strand of DNA will adopt a
conformation that is uniquely dependent on its sequence
conformation. This conformation usually will be different, even if
only a single base is changed. Most conformations have been
reported to alter the physical configuration or size sufficiently
to be detectable by electrophoresis. A number of protocols have
been described for SSCP including, but not limited to, Lee et al.,
Anal. Biochem. 205:289-293 (1992), the entirety of which is herein
incorporated by reference; Suzuki et al., Anal. Biochem. 192:82-84
(1991), the entirety of which is herein incorporated by reference;
Lo et al., Nucleic Acids Research 20:1005-1009 (1992), the entirety
of which is herein incorporated by reference; Sarkar et al.,
Genomics 13:441-443 (1992), the entirety of which is herein
incorporated by reference. It is understood that one or more of the
nucleic acids of the present invention, may be utilized as markers
or probes to detect polymorphisms by SSCP analysis.
[0199] Polymorphisms may also be found using a DNA fingerprinting
technique called amplified fragment length polymorphism (AFLP),
which is based on the selective PCR amplification of restriction
fragments from a total digest of genomic DNA to profile that DNA
(Vos et al., Nucleic Acids Res. 23:4407-4414 (1995), the entirety
of which is herein incorporated by reference). This method allows
for the specific co-amplification of high numbers of restriction
fragments, which can be visualized by PCR without knowledge of the
nucleic acid sequence.
[0200] AFLP employs basically three steps. Initially, a sample of
genomic DNA is cut with restriction enzymes and oligonucleotide
adapters are ligated to the restriction fragments of the DNA. The
restriction fragments are then amplified using PCR by using the
adapter and restriction sequence as target sites for primer
annealing. The selective amplification is achieved by the use of
primers that extend into the restriction fragments, amplifying only
those fragments in which the primer extensions match the nucleotide
flanking the restriction sites. These amplified fragments are then
visualized on a denaturing polyacrylamide gel.
[0201] AFLP analysis has been performed on Salix (Beismann et al.,
Mol. Ecol. 6:989-993 (1997), the entirety of which is herein
incorporated by reference), Acinetobacter (Janssen et al., Int. J.
Syst. Bacteriol. 47:1179-1187 (1997), the entirety of which is
herein incorporated by reference), Aeromonas popoffi (Huys et al.,
Int. J. Syst. Bacteriol. 47:1165-1171 (1997), the entirety of which
is herein incorporated by reference), rice (McCouch et al., Plant
Mol. Biol. 35:89-99 (1997), the entirety of which is herein
incorporated by reference; Nandi et al., Mol. Gen. Genet. 255:1-8
(1997), the entirety of which is herein incorporated by reference;
Cho et al., Genome 39:373-378 (1996), the entirety of which is
herein incorporated by reference), barley (Hordeum vulgare)(Simons
et al., Genomics 44:61-70 (1997), the entirety of which is herein
incorporated by reference; Waugh et al., Mol. Gen. Genet.
255:311-321 (1997), the entirety of which is herein incorporated by
reference; Qi et al., Mol. Gen. Genet. 254:330-336 (1997), the
entirety of which is herein incorporated by reference; Becker et
al., Mol. Gen. Genet. 249:65-73 (1995), the entirety of which is
herein incorporated by reference), potato (Van der Voort et al.,
Mol. Gen. Genet. 255:438-447 (1997), the entirety of which is
herein incorporated by reference; Meksem et al., Mol. Gen. Genet.
249:74-81 (1995), the entirety of which is herein incorporated by
reference), Phytophthora infestans (Van der Lee et al., Fungal
Genet. Biol. 21:278-291 (1997), the entirety of which is herein
incorporated by reference), Bacillus anthracis (Keim et al., J.
Bacteriol. 179:818-824 (1997), the entirety of which is herein
incorporated by reference), Astragalus cremnophylax (Travis et al.,
Mol. Ecol. 5:735-745 (1996), the entirety of which is herein
incorporated by reference), Arabidopsis (Cnops et al., Mol. Gen.
Genet. 253:32-41 (1996), the entirety of which is herein
incorporated by reference), Escherichia coli (Lin et al., Nucleic
Acids Res. 24:3649-3650 (1996), the entirety of which is herein
incorporated by reference), Aeromonas (Huys et al., Int. J. Syst.
Bacteriol. 46:572-580 (1996), the entirety of which is herein
incorporated by reference), nematode (Folkertsma et al., Mol.
Plant. Microbe Interact. 9:47-54 (1996), the entirety of which is
herein incorporated by reference), tomato (Thomas et al., Plant J.
8:785-794 (1995), the entirety of which is herein incorporated by
reference) and human (Latorra et al., PCR Methods Appl. 3:351-358
(1994), the entirety of which is herein incorporated by reference).
AFLP analysis has also been used for fingerprinting mRNA (Money et
al., Nucleic Acids Res. 24:2616-2617 (1996), the entirety of which
is herein incorporated by reference; Bachem et al., Plant
19:745-753 (1996), the entirety of which is herein incorporated by
reference). It is understood that one or more of the nucleic acids
of the present invention, may be utilized as markers or probes to
detect polymorphisms by AFLP analysis or for fingerprinting
RNA.
[0202] Polymorphisms may also be found using random amplified
polymorphic DNA (RAPD) (Williams et al., Nucl. Acids Res.
18:6531-6535 (1990), the entirety of which is herein incorporated
by reference) and cleaveable amplified polymorphic sequences (CAPS)
(Lyamichev et al., Science 260:778-783 (1993), the entirety of
which is herein incorporated by reference). It is understood that
one or more of the nucleic acid molecules of the present invention,
may be utilized as markers or probes to detect polymorphisms by
RAPD or CAPS analysis.
[0203] Through genetic mapping, a fine scale linkage map can be
developed using DNA markers and, then, a genomic DNA library of
large-sized fragments can be screened with molecular markers linked
to the desired trait. Molecular markers are advantageous for
agronomic traits that are otherwise difficult to tag, such as
resistance to pathogens, insects and nematodes, tolerance to
abiotic stress, quality parameters and quantitative traits such as
high yield potential.
[0204] The essential requirements for marker-assisted selection in
a plant breeding program are: (1) the marker(s) should co-segregate
or be closely linked with the desired trait; (2) an efficient means
of screening large populations for the molecular marker(s) should
be available; and (3) the screening technique should have high
reproducibility across laboratories and preferably be economical to
use and be user-friendly.
[0205] The genetic linkage of marker molecules can be established
by a gene mapping model such as, without limitation, the flanking
marker model reported by Lander and Botstein, Genetics 121:185-199
(1989) and the interval mapping, based on maximum likelihood
methods described by Lander and Botstein, Genetics 121:185-199
(1989) and implemented in the software package MAPMAKER/QTL
(Lincoln and Lander, Mapping Genes Controlling Quantitative Traits
Using MAPMAKER/QTL, Whitehead Institute for Biomedical Research,
Massachusetts, (1990). Additional software includes Qgene, Version
2.23 (1996), Department of Plant Breeding and Biometry, 266 Emerson
Hall, Cornell University, Ithaca, N.Y., the manual of which is
herein incorporated by reference in its entirety). Use of Qgene
software is a particularly preferred approach.
[0206] A maximum likelihood estimate (MLE) for the presence of a
marker is calculated, together with an MLE assuming no QTL effect,
to avoid false positives. A log.sub.10 of an odds ratio (LOD) is
then calculated as: LOD=log.sub.10, (MLE for the presence of a
QTL/MLE given no linked QTL).
[0207] The LOD score essentially indicates how much more likely the
data are to have arisen assuming the presence of a QTL than in its
absence. The LOD threshold value for avoiding a false positive with
a given confidence, say 95%, depends on the number of markers and
the length of the genome. Graphs indicating LOD thresholds are set
forth in Lander and Botstein, Genetics 121:185-199 (1989) the
entirety of which is herein incorporated by reference and further
described by Ards and Moreno-Gonzalez, Plant Breeding, Hayward et
al., (eds.) Chapman & Hall, London, pp. 314-331 (1993), the
entirety of which is herein incorporated by reference.
[0208] Additional models can be used. Many modifications and
alternative approaches to interval mapping have been reported,
including the use non-parametric methods (Kruglyak and Lander,
Genetics 139:1421-1428 (1995), the entirety of which is herein
incorporated by reference). Multiple regression methods or models
can be also be used, in which the trait is regressed on a large
number of markers (Jansen, Biometrics in Plant Breeding, van Oijen
and Jansen (eds.), Proceedings of the Ninth Meeting of the Eucarpia
Section Biometrics in Plant Breeding, The Netherlands, pp. 116-124
(1994); Weber and Wricke, Advances in Plant Breeding, Blackwell,
Berlin, 16 (1994), both of which is herein incorporated by
reference in their entirety). Procedures combining interval mapping
with regression analysis, whereby the phenotype is regressed onto a
single putative QTL at a given marker interval and at the same time
onto a number of markers that serve as `cofactors,` have been
reported by Jansen and Stam, Genetics 136:1447-1455 (1994), the
entirety of which is herein incorporated by reference and Zeng,
Genetics 136:1457-1468 (1994) the entirety of which is herein
incorporated by reference. Generally, the use of cofactors reduces
the bias and sampling error of the estimated QTL positions (Utz and
Melchinger, Biometrics in Plant Breeding, van Oijen and Jansen
(eds.) Proceedings of the Ninth Meeting of the Eucarpia Section
Biometrics in Plant Breeding, The Netherlands, pp. 195-204 (1994),
the entirety of which is herein incorporated by reference, thereby
improving the precision and efficiency of QTL mapping (Zeng,
Genetics 136:1457-1468 (1994)). These models can be extended to
multi-environment experiments to analyze genotype-environment
interactions (Jansen et al., Theo. Appl. Genet. 91:33-37 (1995),
the entirety of which is herein incorporated by reference).
[0209] Selection of an appropriate mapping populations is important
to map construction. The choice of appropriate mapping population
depends on the type of marker systems employed (Tanksley et al.,
Molecular mapping plant chromosomes. Chromosome structure and
function: Impact of new concepts, Gustafson and Appels (eds.),
Plenum Press, New York, pp 157-173 (1988), the entirety of which is
herein incorporated by reference). Consideration must be given to
the source of parents (adapted vs. exotic) used in the mapping
population. Chromosome pairing and recombination rates can be
severely disturbed (suppressed) in wide crosses
(adapted.times.exotic) and generally yield greatly reduced linkage
distances. Wide crosses will usually provide segregating
populations with a relatively large array of polymorphisms when
compared to progeny in a narrow cross (adapted.times.adapted).
[0210] An F.sub.2 population is the first generation of selfing
after the hybrid seed is produced. Usually a single F.sub.1 plant
is selfed to generate a population segregating for all the genes in
Mendelian (1:2:1) fashion. Maximum genetic information is obtained
from a completely classified F.sub.2 population using a codominant
marker system (Mather, Measurement of Linkage in Heredity, Methuen
and Co., (1938), the entirety of which is herein incorporated by
reference). In the case of dominant markers, progeny tests (e.g.
F.sub.3, BCF.sub.2) are required to identify the heterozygotes,
thus making it equivalent to a completely classified F.sub.2
population. However, this procedure is often prohibitive because of
the cost and time involved in progeny testing. Progeny testing of
F.sub.2 individuals is often used in map construction where
phenotypes do not consistently reflect genotype (e.g. disease
resistance) or where trait expression is controlled by a QTL.
Segregation data from progeny test populations (e.g. F.sub.3 or
BCF.sub.2) can be used in map construction. Marker-assisted
selection can then be applied to cross progeny based on
marker-trait map associations (F.sub.2, F.sub.3), where linkage
groups have not been completely disassociated by recombination
events (i.e., maximum disequillibrium).
[0211] Recombinant inbred lines (RIL) (genetically related lines;
usually >F.sub.5, developed from continuously selfing F.sub.2
lines towards homozygosity) can be used as a mapping population.
Information obtained from dominant markers can be maximized by
using RIL because all loci are homozygous or nearly so. Under
conditions of tight linkage (i.e., about <10% recombination),
dominant and co-dominant markers evaluated in RIL populations
provide more information per individual than either marker type in
backcross populations (Reiter et al., Proc. Natl. Acad. Sci.
(U.S.A.) 89:1477-1481 (1992), the entirety of which is herein
incorporated by reference). However, as the distance between
markers becomes larger (i.e., loci become more independent), the
information in RIL populations decreases dramatically when compared
to codominant markers.
[0212] Backcross populations (e.g., generated from a cross between
a successful variety (recurrent parent) and another variety (donor
parent) carrying a trait not present in the former) can be utilized
as a mapping population. A series of backcrosses to the recurrent
parent can be made to recover most of its desirable traits. Thus a
population is created consisting of individuals nearly like the
recurrent parent but each individual carries varying amounts or
mosaic of genomic regions from the donor parent. Backcross
populations can be useful for mapping dominant markers if all loci
in the recurrent parent are homozygous and the donor and recurrent
parent have contrasting polymorphic marker alleles (Reiter et al.,
Proc. Natl. Acad. Sci. (U.S.A.) 89:1477-1481 (1992)). Information
obtained from backcross populations using either codominant or
dominant markers is less than that obtained from F.sub.2
populations because one, rather than two, recombinant gametes are
sampled per plant. Backcross populations, however, are more
informative (at low marker saturation) when compared to RILs as the
distance between linked loci increases in RIL populations (i.e.
about 15% recombination). Increased recombination can be beneficial
for resolution of tight linkages, but may be undesirable in the
construction of maps with low marker saturation.
[0213] Near-isogenic lines (NIL) created by many backcrosses to
produce an array of individuals that are nearly identical in
genetic composition except for the trait or genomic region under
interrogation can be used as a mapping population. In mapping with
NILs, only a portion of the polymorphic loci are expected to map to
a selected region.
[0214] Bulk segregant analysis (BSA) is a method developed for the
rapid identification of linkage between markers and traits of
interest (Michelmore et al., Proc. Natl. Acad. Sci. (U.S.A.)
88:9828-9832 (1991), the entirety of which is herein incorporated
by reference). In BSA, two bulked DNA samples are drawn from a
segregating population originating from a single cross. These bulks
contain individuals that are identical for a particular trait
(resistant or susceptible to particular disease) or genomic region
but arbitrary at unlinked regions (i.e. heterozygous). Regions
unlinked to the target region will not differ between the bulked
samples of many individuals in BSA.
[0215] It is understood that one or more of the nucleic acid
molecules of the present invention may be used as molecular
markers. It is also understood that one or more of the protein
molecules of the present invention may be used as molecular
markers.
[0216] In accordance with this aspect of the present invention, a
sample nucleic acid is obtained from plants cells or tissues. Any
source of nucleic acid may be used. Preferably, the nucleic acid is
genomic DNA. The nucleic acid is subjected to restriction
endonuclease digestion. For example, one or more nucleic acid
molecule or fragment thereof of the present invention can be used
as a probe in accordance with the above-described polymorphic
methods. The polymorphism obtained in this approach can then be
cloned to identify the mutation at the coding region which alters
the protein's structure or regulatory region of the gene which
affects its expression level.
[0217] In an aspect of the present invention, one or more of the
nucleic molecules of the present invention are used to determine
the level (i.e., the concentration of mRNA in a sample, etc.) in a
plant (preferably maize or soybean) or pattern (i.e., the kinetics
of expression, rate of decomposition, stability profile, etc.) of
the expression of a protein encoded in part or whole by one or more
of the nucleic acid molecule of the present invention
(collectively, the "Expression Response" of a cell or tissue). As
used herein, the Expression Response manifested by a cell or tissue
is said to be "altered" if it differs from the Expression Response
of cells or tissues of plants not exhibiting the phenotype. To
determine whether a Expression Response is altered, the Expression
Response manifested by the cell or tissue of the plant exhibiting
the phenotype is compared with that of a similar cell or tissue
sample of a plant not exhibiting the phenotype. As will be
appreciated, it is not necessary to re-determine the Expression
Response of the cell or tissue sample of plants not exhibiting the
phenotype each time such a comparison is made; rather, the
Expression Response of a particular plant may be compared with
previously obtained values of normal plants. As used herein, the
phenotype of the organism is any of one or more characteristics of
an organism (e.g. disease resistance, pest tolerance, environmental
tolerance such as tolerance to abiotic stress, male sterility,
quality improvement or yield etc.). A change in genotype or
phenotype may be transient or permanent. Also as used herein, a
tissue sample is any sample that comprises more than one cell. In a
preferred aspect, a tissue sample comprises cells that share a
common characteristic (e.g. derived from root, seed, flower, leaf,
stem or pollen etc.).
[0218] In one aspect of the present invention, an evaluation can be
conducted to determine whether a particular mRNA molecule is
present. One or more of the nucleic acid molecules of the present
invention, preferably one or more of the EST nucleic acid molecules
or fragments thereof of the present invention are utilized to
detect the presence or quantity of the mRNA species. Such molecules
are then incubated with cell or tissue extracts of a plant under
conditions sufficient to permit nucleic acid hybridization. The
detection of double-stranded probe-mRNA hybrid molecules is
indicative of the presence of the mRNA; the amount of such hybrid
formed is proportional to the amount of mRNA. Thus, such probes may
be used to ascertain the level and extent of the mRNA production in
a plant's cells or tissues. Such nucleic acid hybridization may be
conducted under quantitative conditions (thereby providing a
numerical value of the amount of the mRNA present). Alternatively,
the assay may be conducted as a qualitative assay that indicates
either that the mRNA is present, or that its level exceeds a user
set, predefined value.
[0219] A principle of in situ hybridization is that a labeled,
single-stranded nucleic acid probe will hybridize to a
complementary strand of cellular DNA or RNA and, under the
appropriate conditions, these molecules will form a stable hybrid.
When nucleic acid hybridization is combined with histological
techniques, specific DNA or RNA sequences can be identified within
a single cell. An advantage of in situ hybridization over more
conventional techniques for the detection of nucleic acids is that
it allows an investigator to determine the precise spatial
population (Angerer et al., Dev. Biol. 101:477-484 (1984), the
entirety of which is herein incorporated by reference; Angerer et
al., Dev. Biol. 112:157-166 (1985), the entirety of which is herein
incorporated by reference; Dixon et al., EMBO J. 10:1317-1324
(1991), the entirety of which is herein incorporated by reference).
In situ hybridization may be used to measure the steady-state level
of RNA accumulation. It is a sensitive technique and RNA sequences
present in as few as 5-10 copies per cell can be detected (Hardin
et al., J. Mol. Biol. 202:417-431 (1989), the entirety of which is
herein incorporated by reference). A number of protocols have been
devised for in situ hybridization, each with tissue preparation,
hybridization and washing conditions (Meyerowitz, Plant Mol. Biol.
Rep. 5:242-250 (1987), the entirety of which is herein incorporated
by reference; Cox and Goldberg, In: Plant Molecular Biology: A
Practical Approach, Shaw (ed.), pp 1-35, IRL Press, Oxford (1988),
the entirety of which is herein incorporated by reference; Raikhel
et al., In situ RNA hybridization in plant tissues, In: Plant
Molecular Biology Manual, vol. B9:1-32, Kluwer Academic Publisher,
Dordrecht, Belgium (1989), the entirety of which is herein
incorporated by reference).
[0220] In situ hybridization also allows for the localization of
proteins within a tissue or cell (Wilkinson, In Situ Hybridization,
Oxford University Press, Oxford (1992), the entirety of which is
herein incorporated by reference; Langdale, In Situ Hybridization
In: The Maize Handbook, Freeling and Walbot (eds.), pp 165-179,
Springer-Verlag, New York (1994), the entirety of which is herein
incorporated by reference). It is understood that one or more of
the molecules of the present invention, preferably one or more of
the EST nucleic acid molecules or fragments thereof of the present
invention or one or more of the antibodies of the present invention
may be utilized to detect the level or pattern of a methionine
pathway protein or mRNA thereof by in situ hybridization.
[0221] Fluorescent in situ hybridization allows the localization of
a particular DNA sequence along a chromosome which is useful, among
other uses, for gene mapping, following chromosomes in hybrid lines
or detecting chromosomes with translocations, transversions or
deletions. In situ hybridization has been used to identify
chromosomes in several plant species (Griffor et al., Plant Mol.
Biol. 17:101-109 (1991), the entirety of which is herein
incorporated by reference; Gustafson et al., Proc. Natl. Acad. Sci.
(U.S.A.) 87:1899-1902 (1990), herein incorporated by reference;
Mukai and Gill, Genome 34:448-452 (1991), the entirety of which is
herein incorporated by reference; Schwarzacher and Heslop-Harrison,
Genome 34:317-323 (1991); Wang et al., Jpn. J. Genet. 66:313-316
(1991), the entirety of which is herein incorporated by reference;
Parra and Windle, Nature Genetics 5:17-21 (1993), the entirety of
which is herein incorporated by reference). It is understood that
the nucleic acid molecules of the present invention may be used as
probes or markers to localize sequences along a chromosome.
[0222] Another method to localize the expression of a molecule is
tissue printing. Tissue printing provides a way to screen, at the
same time on the same membrane many tissue sections from different
plants or different developmental stages. Tissue-printing
procedures utilize films designed to immobilize proteins and
nucleic acids. In essence, a freshly cut section of a tissue is
pressed gently onto nitrocellulose paper, nylon membrane or
polyvinylidene difluoride membrane. Such membranes are commercially
available (e.g. Millipore, Bedford, Mass. U.S.A.). The contents of
the cut cell transfer onto the membrane and the contents and are
immobilized to the membrane. The immobilized contents form a latent
print that can be visualized with appropriate probes. When a plant
tissue print is made on nitrocellulose paper, the cell walls leave
a physical print that makes the anatomy visible without further
treatment (Varner and Taylor, Plant Physiol. 91:31-33 (1989), the
entirety of which is herein incorporated by reference).
[0223] Tissue printing on substrate films is described by Daoust,
Exp. Cell Res. 12:203-211 (1957), the entirety of which is herein
incorporated by reference, who detected amylase, protease,
ribonuclease and deoxyribonuclease in animal tissues using starch,
gelatin and agar films. These techniques can be applied to plant
tissues (Yomo and Taylor, Planta 112:35-43 (1973); the entirety of
which is herein incorporated by reference; Harris and Chrispeels,
Plant Physiol. 56:292-299 (1975), the entirety of which is herein
incorporated by reference). Advances in membrane technology have
increased the range of applications of Daoust's tissue-printing
techniques allowing (Cassab and Varner, J. Cell. Biol.
105:2581-2588 (1987), the entirety of which is herein incorporated
by reference) the histochemical localization of various plant
enzymes and deoxyribonuclease on nitrocellulose paper and nylon
(Spruce et al., Phytochemistry 26:2901-2903 (1987), the entirety of
which is herein incorporated by reference; Barres et al., Neuron
5:527-544 (1990), the entirety of which is herein incorporated by
reference; Reid and Pont-Lezica, Tissue Printing: Tools for the
Study of Anatomy, Histochemistry and Gene Expression, Academic
Press, New York, N.Y. (1992), the entirety of which is herein
incorporated by reference; Reid et al., Plant Physiol. 93:160-165
(1990), the entirety of which is herein incorporated by reference;
Ye et al., Plant J. 1:175-183 (1991), the entirety of which is
herein incorporated by reference).
[0224] It is understood that one or more of the molecules of the
present invention, preferably one or more of the EST nucleic acid
molecules or fragments thereof of the present invention or one or
more of the antibodies of the present invention may be utilized to
detect the presence or quantity of a methionine pathway protein by
tissue printing.
[0225] Further it is also understood that any of the nucleic acid
molecules of the present invention may be used as marker nucleic
acids and or probes in connection with methods that require probes
or marker nucleic acids. As used herein, a probe is an agent that
is utilized to determine an attribute or feature (e.g. presence or
absence, location, correlation, etc.) of a molecule, cell, tissue
or plant. As used herein, a marker nucleic acid is a nucleic acid
molecule that is utilized to determine an attribute or feature
(e.g., presence or absence, location, correlation, etc.) or a
molecule, cell, tissue or plant.
[0226] A microarray-based method for high-throughput monitoring of
plant gene expression may be utilized to measure gene-specific
hybridization targets. This `chip`-based approach involves using
microarrays of nucleic acid molecules as gene-specific
hybridization targets to quantitatively measure expression of the
corresponding plant genes (Schena et al., Science 270:467-470
(1995), the entirety of which is herein incorporated by reference;
Shalon, Ph.D. Thesis, Stanford University (1996), the entirety of
which is herein incorporated by reference). Every nucleotide in a
large sequence can be queried at the same time. Hybridization can
be used to efficiently analyze nucleotide sequences.
[0227] Several microarray methods have been described. One method
compares the sequences to be analyzed by hybridization to a set of
oligonucleotides representing all possible subsequences (Bains and
Smith, J. Theor. Biol. 135:303-307 (1989), the entirety of which is
herein incorporated by reference). A second method hybridizes the
sample to an array of oligonucleotide or cDNA molecules. An array
consisting of oligonucleotides complementary to subsequences of a
target sequence can be used to determine the identity of a target
sequence, measure its amount and detect differences between the
target and a reference sequence. Nucleic acid molecules microarrays
may also be screened with protein molecules or fragments thereof to
determine nucleic acid molecules that specifically bind protein
molecules or fragments thereof.
[0228] The microarray approach may be used with polypeptide targets
(U.S. Pat. No. 5,445,934; U.S. Pat. No. 5,143,854; U.S. Pat. No.
5,079,600; U.S. Pat. No. 4,923,901, all of which are herein
incorporated by reference in their entirety). Essentially,
polypeptides are synthesized on a substrate (microarray) and these
polypeptides can be screened with either protein molecules or
fragments thereof or nucleic acid molecules in order to screen for
either protein molecules or fragments thereof or nucleic acid
molecules that specifically bind the target polypeptides. (Fodor et
al., Science 251:767-773 (1991), the entirety of which is herein
incorporated by reference). It is understood that one or more of
the nucleic acid molecules or protein or fragments thereof of the
present invention may be utilized in a microarray based method.
[0229] In a preferred embodiment of the present invention
microarrays may be prepared that comprise nucleic acid molecules
where such nucleic acid molecules encode at least one, preferably
at least two, more preferably at least three, even more preferably
at least four, five six or seven methionine pathway enzymes. In a
preferred embodiment the nucleic acid molecules are selected from
the group consisting of a nucleic acid molecule that encodes a
maize or a soybean methionine adenosyltransferase enzyme or
fragment thereof, a nucleic acid molecule that encodes a maize or a
soybean S-adenosylmethionine decarboxylase enzyme or fragment
thereof, a nucleic acid molecule that encodes a maize or a soybean
aspartate kinase enzyme or fragment thereof, a nucleic acid
molecule that encodes a maize or a soybean aspartate-semialdehyde
dehydrogenase enzyme or fragment thereof, a nucleic acid molecule
that encodes a maize or a soybean O-succinylhomoserine
(thiol)-lyase enzyme or fragment thereof, a nucleic acid molecule
that encodes a maize or a soybean cystathionine .beta.-lyase enzyme
or fragment thereof, a nucleic acid molecule that encodes a maize
or a soybean
5-methyltetrahydropteroyltriglutamate-homocysteine-S-methyltransferase
enzyme or fragment thereof, a nucleic acid molecule that encodes a
maize or a soybean adenosylhomocysteinase enzyme or fragment
thereof, a nucleic acid molecule that encodes a maize or a soybean
cystathionine .beta.-synthase enzyme or fragment thereof, a nucleic
acid molecule that encodes a maize or a soybean cystathionine
.gamma.-lyase enzyme or fragment thereof and a nucleic acid
molecule that encodes a maize or a soybean O-acetylhomoserine
(thiol)-lyase enzyme or fragment thereof.
[0230] Site directed mutagenesis may be utilized to modify nucleic
acid sequences, particularly as it is a technique that allows one
or more of the amino acids encoded by a nucleic acid molecule to be
altered (e.g. a threonine to be replaced by a methionine). Three
basic methods for site directed mutagenesis are often employed.
These are cassette mutagenesis (Wells et al., Gene 34:315-323
(1985), the entirety of which is herein incorporated by reference),
primer extension (Gilliam et al., Gene 12:129-137 (1980), the
entirety of which is herein incorporated by reference; Zoller and
Smith, Methods Enzymol. 100:468-500 (1983), the entirety of which
is herein incorporated by reference; Dalbadie-McFarland et al.,
Proc. Natl. Acad. Sci. (U.S.A.) 79:6409-6413 (1982), the entirety
of which is herein incorporated by reference) and methods based
upon PCR (Scharf et al., Science 233:1076-1078 (1986), the entirety
of which is herein incorporated by reference; Higuchi et al.,
Nucleic Acids Res. 16:7351-7367 (1988), the entirety of which is
herein incorporated by reference). Site directed mutagenesis
approaches are also described in European Patent 0 385 962, the
entirety of which is herein incorporated by reference; European
Patent 0 359 472, the entirety of which is herein incorporated by
reference; and PCT Patent Application WO 93/07278, the entirety of
which is herein incorporated by reference.
[0231] Site directed mutagenesis strategies have been applied to
plants for both in vitro as well as in vivo site directed
mutagenesis (Lanz et al., J. Biol. Chem. 266:9971-9976 (1991), the
entirety of which is herein incorporated by reference; Kovgan and
Zhdanov, Biotekhnologiya 5:148-154, No. 207160n, Chemical Abstracts
110:225 (1989), the entirety of which is herein incorporated by
reference; Ge et al., Proc. Natl. Acad. Sci. (U.S.A.) 86:4037-4041
(1989), the entirety of which is herein incorporated by reference;
Zhu et al., J. Biol. Chem. 271:18494-18498 (1996), the entirety of
which is herein incorporated by reference; Chu et al., Biochemistry
33:6150-6157 (1994), the entirety of which is herein incorporated
by reference; Small et al., EMBO J. 11:1291-1296 (1992), the
entirety of which is herein incorporated by reference; Cho et al.,
Mol. Biotechnol. 8:13-16 (1997), the entirety of which is herein
incorporated by reference; Kita et al., J. Biol. Chem.
271:26529-26535 (1996), the entirety of which is herein
incorporated by reference, Jin et al., Mol. Microbiol. 7:555-562
(1993), the entirety of which is herein incorporated by reference;
Hatfield and Vierstra, J. Biol. Chem. 267:14799-14803 (1992), the
entirety of which is herein incorporated by reference; Zhao et al.,
Biochemistry 31:5093-5099 (1992), the entirety of which is herein
incorporated by reference).
[0232] Any of the nucleic acid molecules of the present invention
may either be modified by site directed mutagenesis or used as, for
example, nucleic acid molecules that are used to target other
nucleic acid molecules for modification. It is understood that
mutants with more than one altered nucleotide can be constructed
using techniques that practitioners are familiar with such as
isolating restriction fragments and ligating such fragments into an
expression vector (see, for example, Sambrook et al., Molecular
Cloning: A Laboratory Manual, Cold Spring Harbor Press (1989)).
[0233] Sequence-specific DNA-binding proteins play a role in the
regulation of transcription. The isolation of recombinant cDNAs
encoding these proteins facilitates the biochemical analysis of
their structural and functional properties. Genes encoding such
DNA-binding proteins have been isolated using classical genetics
(Vollbrecht et al., Nature 350: 241-243 (1991), the entirety of
which is herein incorporated by reference) and molecular
biochemical approaches, including the screening of recombinant cDNA
libraries with antibodies (Landschulz et al., Genes Dev. 2:786-800
(1988), the entirety of which is herein incorporated by reference)
or DNA probes (Bodner et al., Cell 55:505-518 (1988), the entirety
of which is herein incorporated by reference). In addition, an in
situ screening procedure has been used and has facilitated the
isolation of sequence-specific DNA-binding proteins from various
plant species (Gilmartin et al., Plant Cell 4:839-849 (1992), the
entirety of which is herein incorporated by reference; Schindler et
al., EMBO J. 11:1261-1273 (1992), the entirety of which is herein
incorporated by reference). An in situ screening protocol does not
require the purification of the protein of interest (Vinson et al.,
Genes Dev. 2:801-806 (1988), the entirety of which is herein
incorporated by reference; Singh et al., Cell 52:415-423 (1988),
the entirety of which is herein incorporated by reference).
[0234] Two steps may be employed to characterize DNA-protein
interactions. The first is to identify promoter fragments that
interact with DNA-binding proteins, to titrate binding activity, to
determine the specificity of binding and to determine whether a
given DNA-binding activity can interact with related DNA sequences
(Sambrook et al., Molecular Cloning: A Laboratory Manual, 2.sup.nd
edition, Cold Spring Harbor Laboratory Press, Cold Spring Harbor,
N.Y. (1989)). Electrophoretic mobility-shift assay is a widely used
assay. The assay provides a rapid and sensitive method for
detecting DNA-binding proteins based on the observation that the
mobility of a DNA fragment through a nondenaturing, low-ionic
strength polyacrylamide gel is retarded upon association with a
DNA-binding protein (Fried and Crother, Nucleic Acids Res.
9:6505-6525 (1981), the entirety of which is herein incorporated by
reference). When one or more specific binding activities have been
identified, the exact sequence of the DNA bound by the protein may
be determined. Several procedures for characterizing
protein/DNA-binding sites are used, including methylation and
ethylation interference assays (Maxam and Gilbert, Methods Enzymol.
65:499-560 (1980), the entirety of which is herein incorporated by
reference; Wissman and Hillen, Methods Enzymol. 208:365-379 (1991),
the entirety of which is herein incorporated by reference),
footprinting techniques employing DNase I (Galas and Schmitz,
Nucleic Acids Res. 5:3157-3170 (1978), the entirety of which is
herein incorporated by reference), 1,10-phenanthroline-copper ion
methods (Sigman et al., Methods Enzymol. 208:414-433 (1991), the
entirety of which is herein incorporated by reference) and hydroxyl
radicals methods (Dixon et al., Methods Enzymol. 208:414-433
(1991), the entirety of which is herein incorporated by reference).
It is understood that one or more of the nucleic acid molecules of
the present invention may be utilized to identify a protein or
fragment thereof that specifically binds to a nucleic acid molecule
of the present invention. It is also understood that one or more of
the protein molecules or fragments thereof of the present invention
may be utilized to identify a nucleic acid molecule that
specifically binds to it.
[0235] A two-hybrid system is based on the fact that many cellular
functions are carried out by proteins, such as transcription
factors, that interact (physically) with one another. Two-hybrid
systems have been used to probe the function of new proteins (Chien
et al., Proc. Natl. Acad. Sci. (U.S.A.) 88:9578-9582 (1991) the
entirety of which is herein incorporated by reference; Durfee et
al., Genes Dev. 7:555-569 (1993) the entirety of which is herein
incorporated by reference; Choi et al., Cell 78:499-512 (1994), the
entirety of which is herein incorporated by reference; Kranz et
al., Genes Dev. 8:313-327 (1994), the entirety of which is herein
incorporated by reference).
[0236] Interaction mating techniques have facilitated a number of
two-hybrid studies of protein-protein interaction. Interaction
mating has been used to examine interactions between small sets of
tens of proteins (Finley and Brent, Proc. Natl. Acad. Sci. (U.S.A.)
91:12098-12984 (1994), the entirety of which is herein incorporated
by reference), larger sets of hundreds of proteins (Bendixen et
al., Nucl. Acids Res. 22:1778-1779 (1994), the entirety of which is
herein incorporated by reference) and to comprehensively map
proteins encoded by a small genome (Bartel et al., Nature Genetics
12:72-77 (1996), the entirety of which is herein incorporated by
reference). This technique utilizes proteins fused to the
DNA-binding domain and proteins fused to the activation domain.
They are expressed in two different haploid yeast strains of
opposite mating type and the strains are mated to determine if the
two proteins interact. Mating occurs when haploid yeast strains
come into contact and result in the fusion of the two haploids into
a diploid yeast strain. An interaction can be determined by the
activation of a two-hybrid reporter gene in the diploid strain. An
advantage of this technique is that it reduces the number of yeast
transformations needed to test individual interactions. It is
understood that the protein-protein interactions of protein or
fragments thereof of the present invention may be investigated
using the two-hybrid system and that any of the nucleic acid
molecules of the present invention that encode such proteins or
fragments thereof may be used to transform yeast in the two-hybrid
system.
[0237] (a) Plant Constructs and Plant Transformants
[0238] One or more of the nucleic acid molecules of the present
invention may be used in plant transformation or transfection.
Exogenous genetic material may be transferred into a plant cell and
the plant cell regenerated into a whole, fertile or sterile plant.
Exogenous genetic material is any genetic material, whether
naturally occurring or otherwise, from any source that is capable
of being inserted into any organism. Such genetic material may be
transferred into either monocotyledons and dicotyledons including,
but not limited to maize (pp 63-69), soybean (pp 50-60),
Arabidopsis (p 45), phaseolus (pp 47-49), peanut (pp 49-50),
alfalfa (p 60), wheat (pp 69-71), rice (pp 72-79), oat (pp 80-81),
sorghum (p 83), rye (p 84), tritordeum (p 84), millet (p 85),
fescue (p 85), perennial ryegrass (p 86), sugarcane (p 87),
cranberry (p 101), papaya (pp 101-102), banana (p 103), banana (p
103), muskmelon (p 104), apple (p 104), cucumber (p 105),
dendrobium (p 109), gladiolus (p 110), chrysanthemum (p 110),
liliacea (p 111), cotton (pp 113-114), eucalyptus (p 115),
sunflower (p 118), canola (p 118), turfgrass (p 121), sugarbeet (p
122), coffee (p 122) and dioscorea (p 122), (Christou, In: Particle
Bombardment for Genetic Engineering of Plants, Biotechnology
Intelligence Unit. Academic Press, San Diego, Calif. (1996), the
entirety of which is herein incorporated by reference).
[0239] Transfer of a nucleic acid that encodes for a protein can
result in overexpression of that protein in a transformed cell or
transgenic plant. One or more of the proteins or fragments thereof
encoded by nucleic acid molecules of the present invention may be
overexpressed in a transformed cell or transformed plant.
Particularly, any of the methionine pathway proteins or fragments
thereof may be overexpressed in a transformed cell or transgenic
plant. Such overexpression may be the result of transient or stable
transfer of the exogenous genetic material.
[0240] Exogenous genetic material may be transferred into a plant
cell and the plant cell by the use of a DNA vector or construct
designed for such a purpose. Design of such a vector is generally
within the skill of the art (See, Plant Molecular Biology: A
Laboratory Manual, Clark (ed.), Springier, N.Y. (1997), the
entirety of which is herein incorporated by reference).
[0241] A construct or vector may include a plant promoter to
express the protein or protein fragment of choice. A number of
promoters which are active in plant cells have been described in
the literature. These include the nopaline synthase (NOS) promoter
(Ebert et al., Proc. Natl. Acad. Sci. (U.S.A.) 84:5745-5749 (1987),
the entirety of which is herein incorporated by reference), the
octopine synthase (OCS) promoter (which are carried on
tumor-inducing plasmids of Agrobacterium tumefaciens), the
caulimovirus promoters such as the cauliflower mosaic virus (CaMV)
19S promoter (Lawton et al., Plant Mol. Biol. 9:315-324 (1987), the
entirety of which is herein incorporated by reference) and the CAMV
35S promoter (Odell et al., Nature 313:810-812 (1985), the entirety
of which is herein incorporated by reference), the figwort mosaic
virus 35S-promoter, the light-inducible promoter from the small
subunit of ribulose-1,5-bis-phosphate carboxylase (ssRUBISCO), the
Adh promoter (Walker et al., Proc. Natl. Acad. Sci. (U.S.A.)
84:6624-6628 (1987), the entirety of which is herein incorporated
by reference), the sucrose synthase promoter (Yang et al., Proc.
Natl. Acad. Sci. (U.S.A.) 87:4144-4148 (1990), the entirety of
which is herein incorporated by reference), the R gene complex
promoter (Chandler et al., The Plant Cell 1:1175-1183 (1989), the
entirety of which is herein incorporated by reference) and the
chlorophyll a/b binding protein gene promoter, etc. These promoters
have been used to create DNA constructs which have been expressed
in plants; see, e.g., PCT publication WO 84/02913, herein
incorporated by reference in its entirety.
[0242] Promoters which are known or are found to cause
transcription of DNA in plant cells can be used in the present
invention. Such promoters may be obtained from a variety of sources
such as plants and plant viruses. It is preferred that the
particular promoter selected should be capable of causing
sufficient expression to result in the production of an effective
amount of the methionine pathway protein to cause the desired
phenotype. In addition to promoters that are known to cause
transcription of DNA in plant cells, other promoters may be
identified for use in the current invention by screening a plant
cDNA library for genes which are selectively or preferably
expressed in the target tissues or cells.
[0243] For the purpose of expression in source tissues of the
plant, such as the leaf, seed, root or stem, it is preferred that
the promoters utilized in the present invention have relatively
high expression in these specific tissues. For this purpose, one
may choose from a number of promoters for genes with tissue- or
cell-specific or -enhanced expression. Examples of such promoters
reported in the literature include the chloroplast glutamine
synthetase GS2 promoter from pea (Edwards et al., Proc. Natl. Acad.
Sci. (U.S.A.) 87:3459-3463 (1990), herein incorporated by reference
in its entirety), the chloroplast fructose-1,6-biphosphatase
(FBPase) promoter from wheat (Lloyd et al., Mol. Gen. Genet.
225:209-216 (1991), herein incorporated by reference in its
entirety), the nuclear photosynthetic ST-LS1 promoter from potato
(Stockhaus et al., EMBO J. 8:2445-2451 (1989), herein incorporated
by reference in its entirety), the serine/threonine kinase (PAL)
promoter and the glucoamylase (CHS) promoter from Arabidopsis
thaliana. Also reported to be active in photosynthetically active
tissues are the ribulose-1,5-bisphosphate carboxylase (RbcS)
promoter from eastern larch (Larix laricina), the promoter for the
cab gene, cab6, from pine (Yamamoto et al., Plant Cell Physiol.
35:773-778 (1994), herein incorporated by reference in its
entirety), the promoter for the Cab-1 gene from wheat (Fejes et
al., Plant Mol. Biol. 15:921-932 (1990), herein incorporated by
reference in its entirety), the promoter for the CAB-1 gene from
spinach (Lubberstedt et al., Plant Physiol. 104:997-1006 (1994),
herein incorporated by reference in its entirety), the promoter for
the cab1R gene from rice (Luan et al., Plant Cell. 4:971-981
(1992), the entirety of which is herein incorporated by reference),
the pyruvate, orthophosphate dikinase (PPDK) promoter from maize
(Matsuoka et al., Proc. Natl. Acad. Sci. (U.S.A.) 90: 9586-9590
(1993), herein incorporated by reference in its entirety), the
promoter for the tobacco Lhcb1*2 gene (Cerdan et al., Plant Mol.
Biol. 33:245-255 (1997), herein incorporated by reference in its
entirety), the Arabidopsis thaliana SUC2 sucrose-H+symporter
promoter (Truernit et al., Planta. 196:564-570 (1995), herein
incorporated by reference in its entirety) and the promoter for the
thylakoid membrane proteins from spinach (psaD, psaF, psaE, PC,
FNR, atpC, atpD, cab, rbcS). Other promoters for the chlorophyll
a/b-binding proteins may also be utilized in the present invention,
such as the promoters for LhcB gene and PsbP gene from white
mustard (Sinapis alba; Kretsch et al., Plant Mol. Biol. 28:219-229
(1995), the entirety of which is herein incorporated by
reference).
[0244] For the purpose of expression in sink tissues of the plant,
such as the tuber of the potato plant, the fruit of tomato, or the
seed of maize, wheat, rice and barley, it is preferred that the
promoters utilized in the present invention have relatively high
expression in these specific tissues. A number of promoters for
genes with tuber-specific or -enhanced expression are known,
including the class I patatin promoter (Bevan et al., EMBO J.
8:1899-1906 (1986); Jefferson et al., Plant Mol. Biol. 14:995-1006
(1990), both of which are herein incorporated by reference in its
entirety), the promoter for the potato tuber ADPGPP genes, both the
large and small subunits, the sucrose synthase promoter (Salanoubat
and Belliard, Gene. 60:47-56 (1987), Salanoubat and Belliard, Gene.
84:181-185 (1989), both of which are incorporated by reference in
their entirety), the promoter for the major tuber proteins
including the 22 kd protein complexes and proteinase inhibitors
(Hannapel, Plant Physiol. 101:703-704 (1993), herein incorporated
by reference in its entirety), the promoter for the granule bound
starch synthase gene (GBSS) (Visser et al., Plant Mol. Biol.
17:691-699 (1991), herein incorporated by reference in its
entirety) and other class I and II patatins promoters
(Koster-Topfer et al., Mol Gen Genet. 219:390-396 (1989); Mignery
et al., Gene. 62:27-44 (1988), both of which are herein
incorporated by reference in their entirety).
[0245] Other promoters can also be used to express a methionine
pathway protein or fragment thereof in specific tissues, such as
seeds or fruits. The promoter for .beta.-conglycinin (Chen et al.,
Dev. Genet. 10: 112-122 (1989), herein incorporated by reference in
its entirety) or other seed-specific promoters such as the napin
and phaseolin promoters, can be used. The zeins are a group of
storage proteins found in maize endosperm. Genomic clones for zein
genes have been isolated (Pedersen et al., Cell 29:1015-1026
(1982), herein incorporated by reference in its entirety) and the
promoters from these clones, including the 15 kD, 16 kD, 19 kD, 22
kD, 27 kD and .gamma. genes, could also be used. Other promoters
known to function, for example, in maize include the promoters for
the following genes: waxy, Brittle, Shrunken 2, Branching enzymes I
and II, starch synthases, debranching enzymes, oleosins, glutelins
and sucrose synthases. A particularly preferred promoter for maize
endosperm expression is the promoter for the glutelin gene from
rice, more particularly the Osgt-1 promoter (Zheng et al., Mol.
Cell. Biol. 13:5829-5842 (1993), herein incorporated by reference
in its entirety). Examples of promoters suitable for expression in
wheat include those promoters for the ADPglucose pyrosynthase
(ADPGPP) subunits, the granule bound and other starch synthase, the
branching and debranching enzymes, the embryogenesis-abundant
proteins, the gliadins and the glutenins. Examples of such
promoters in rice include those promoters for the ADPGPP subunits,
the granule bound and other starch synthase, the branching enzymes,
the debranching enzymes, sucrose synthases and the glutelins. A
particularly preferred promoter is the promoter for rice glutelin,
Osgt-1. Examples of such promoters for barley include those for the
ADPGPP subunits, the granule bound and other starch synthase, the
branching enzymes, the debranching enzymes, sucrose synthases, the
hordeins, the embryo globulins and the aleurone specific
proteins.
[0246] Root specific promoters may also be used. An example of such
a promoter is the promoter for the acid chitinase gene (Samac et
al., Plant Mol. Biol. 25:587-596 (1994), the entirety of which is
herein incorporated by reference). Expression in root tissue could
also be accomplished by utilizing the root specific subdomains of
the CaMV35S promoter that have been identified (Lam et al., Proc.
Natl. Acad. Sci. (U.S.A.) 86:7890-7894 (1989), herein incorporated
by reference in its entirety). Other root cell specific promoters
include those reported by Conkling et al. (Conkling et al., Plant
Physiol. 93:1203-1211 (1990), the entirety of which is herein
incorporated by reference).
[0247] Additional promoters that may be utilized are described, for
example, in U.S. Pat. Nos. 5,378,619; 5,391,725; 5,428,147;
5,447,858; 5,608,144; 5,608,144; 5,614,399; 5,633,441; 5,633,435;
and 4,633,436, all of which are herein incorporated in their
entirety. In addition, a tissue specific enhancer may be used
(Fromm et al., The Plant Cell 1:977-984 (1989), the entirety of
which is herein incorporated by reference).
[0248] Constructs or vectors may also include with the coding
region of interest a nucleic acid sequence that acts, in whole or
in part, to terminate transcription of that region. For example,
such sequences have been isolated including the Tr7 3' sequence,
and the NOS 3' sequence (Ingelbrecht et al., The Plant Cell
1:671-680 (1989), the entirety of which is herein incorporated by
reference; Bevan et al., Nucleic Acids Res. 11:369-385 (1983), the
entirety of which is herein incorporated by reference), or the
like.
[0249] A vector or construct may also include regulatory elements.
Examples of such include the Adh intron 1 (Callis et al., Genes and
Develop. 1:1183-1200 (1987), the entirety of which is herein
incorporated by reference), the sucrose synthase intron et al.,
Plant Physiol. 91:1575-1579 (1989), the entirety of which is herein
incorporated by reference) and the TMV omega element (Gallie et
al., The Plant Cell 1:301-311 (1989), the entirety of which is
herein incorporated by reference). These and other regulatory
elements may be included when appropriate.
[0250] A vector or construct may also include a selectable marker.
Selectable markers may also be used to select for plants or plant
cells that contain the exogenous genetic material. Examples of such
include, but are not limited to, a neo gene (Potrykus et al., Mol.
Gen. Genet. 199:183-188 (1985), the entirety of which is herein
incorporated by reference) which codes for kanamycin resistance and
can be selected for using kanamycin, G418, etc.; a bar gene which
codes for bialaphos resistance; a mutant EPSP synthase gene
(Hinchee et al., Bio/Technology 6:915-922 (1988), the entirety of
which is herein incorporated by reference) which encodes glyphosate
resistance; a nitrilase gene which confers resistance to bromoxynil
(Stalker et al., J. Biol. Chem. 263:6310-6314 (1988), the entirety
of which is herein incorporated by reference); a mutant
acetolactate synthase gene (ALS) which confers imidazolinone or
sulphonylurea resistance (European Patent Application 154,204 (Sep.
11, 1985), the entirety of which is herein incorporated by
reference); and a methotrexate resistant DHFR gene (Thillet et al.,
J. Biol. Chem. 263:12500-12508 (1988), the entirety of which is
herein incorporated by reference).
[0251] A vector or construct may also include a transit peptide.
Incorporation of a suitable chloroplast transit peptide may also be
employed (European Patent Application Publication Number 0218571,
the entirety of which is herein incorporated by reference).
Translational enhancers may also be incorporated as part of the
vector DNA. DNA constructs could contain one or more 5'
non-translated leader sequences which may serve to enhance
expression of the gene products from the resulting mRNA
transcripts. Such sequences may be derived from the promoter
selected to express the gene or can be specifically modified to
increase translation of the mRNA. Such regions may also be obtained
from viral RNAs, from suitable eukaryotic genes, or from a
synthetic gene sequence. For a review of optimizing expression of
transgenes, see Koziel et al., Plant Mol. Biol. 32:393-405 (1996),
the entirety of which is herein incorporated by reference.
[0252] A vector or construct may also include a screenable marker.
Screenable markers may be used to monitor expression. Exemplary
screenable markers include a .beta.-glucuronidase or uidA gene
(GUS) which encodes an enzyme for which various chromogenic
substrates are known (Jefferson, Plant Mol. Biol, Rep. 5:387-405
(1987), the entirety of which is herein incorporated by reference;
Jefferson et al., EMBO J. 6:3901-3907 (1987), the entirety of which
is herein incorporated by reference); an R-locus gene, which
encodes a product that regulates the production of anthocyanin
pigments (red color) in plant tissues (Dellaporta et al., Stadler
Symposium 11:263-282 (1988), the entirety of which is herein
incorporated by reference); a .beta.-lactamase gene (Sutcliffe et
al., Proc. Natl. Acad. Sci. (U.S.A.) 75:3737-3741 (1978), the
entirety of which is herein incorporated by reference), a gene
which encodes an enzyme for which various chromogenic substrates
are known (e.g., PADAC, a chromogenic cephalosporin); a luciferase
gene (Ow et al., Science 234:856-859 (1986), the entirety of which
is herein incorporated by reference); a xylE gene (Zukowsky et al.,
Proc. Natl. Acad. Sci. (U.S.A.) 80:1101-1105 (1983), the entirety
of which is herein incorporated by reference) which encodes a
catechol diozygenase that can convert chromogenic catechols; an
.alpha.-amylase gene (Ikatu et al., Bio/Technol. 8:241-242 (1990),
the entirety of which is herein incorporated by reference); a
tyrosinase gene (Katz et al., J. Gen. Microbiol. 129:2703-2714
(1983), the entirety of which is herein incorporated by reference)
which encodes an enzyme capable of oxidizing tyrosine to DOPA and
dopaquinone which in turn condenses to melanin; an
.alpha.-galactosidase, which will turn a chromogenic
.alpha.-galactose substrate.
[0253] Included within the terms "selectable or screenable marker
genes" are also genes which encode a secretable marker whose
secretion can be detected as a means of identifying or selecting
for transformed cells. Examples include markers which encode a
secretable antigen that can be identified by antibody interaction,
or even secretable enzymes which can be detected catalytically.
Secretable proteins fall into a number of classes, including small,
diffusible proteins which are detectable, (e.g., by ELISA), small
active enzymes which are detectable in extracellular solution
(e.g., .alpha.-amylase, .beta.-lactamase, phosphinothricin
transferase), or proteins which are inserted or trapped in the cell
wall (such as proteins which include a leader sequence such as that
found in the expression unit of extension or tobacco PR-S). Other
possible selectable and/or screenable marker genes will be apparent
to those of skill in the art.
[0254] There are many methods for introducing transforming nucleic
acid molecules into plant cells. Suitable methods are believed to
include virtually any method by which nucleic acid molecules may be
introduced into a cell, such as by Agrobacterium infection or
direct delivery of nucleic acid molecules such as, for example, by
PEG-mediated transformation, by electroporation or by acceleration
of DNA coated particles, etc (Potrykus, Ann. Rev. Plant Physiol.
Plant Mol. Biol. 42:205-225 (1991), the entirety of which is herein
incorporated by reference; Vasil, Plant Mol. Biol. 25:925-937
(1994), the entirety of which is herein incorporated by reference).
For example, electroporation has been used to transform maize
protoplasts (Fromm et al., Nature 312:791-793 (1986), the entirety
of which is herein incorporated by reference).
[0255] Other vector systems suitable for introducing transforming
DNA into a host plant cell include but are not limited to binary
artificial chromosome (BIBAC) vectors (Hamilton et al., Gene
200:107-116 (1997), the entirety of which is herein incorporated by
reference); and transfection with RNA viral vectors (Della-Cioppa
et al., Ann. N.Y. Acad. Sci. (1996), 792 (Engineering Plants for
Commercial Products and Applications), 57-61, the entirety of which
is herein incorporated by reference). Additional vector systems
also include plant selectable YAC vectors such as those described
in Mullen et al., Molecular Breeding 4:449-457 (1988), the
entireity of which is herein incorporated by reference).
[0256] Technology for introduction of DNA into cells is well known
to those of skill in the art. Four general methods for delivering a
gene into cells have been described: (1) chemical methods (Graham
and van der Eb, Virology, 54:536-539 (1973), the entirety of which
is herein incorporated by reference); (2) physical methods such as
microinjection (Capecchi, Cell 22:479-488 (1980), the entirety of
which is herein incorporated by reference), electroporation (Wong
and Neumann, Biochem. Biophys. Res. Commun. 107:584-587 (1982);
Fromm et al., Proc. Natl. Acad. Sci. (U.S.A.) 82:5824-5828 (1985);
U.S. Pat. No. 5,384,253, all of which are herein incorporated in
their entirety); and the gene gun (Johnston and Tang, Methods Cell
Biol. 43:353-365 (1994), the entirety of which is herein
incorporated by reference); (3) viral vectors (Clapp, Clin.
Perinatol. 20:155-168 (1993); Lu et al., J. Exp. Med. 178:2089-2096
(1993); Eglitis and Anderson, Biotechniques 6:608-614 (1988), all
of which are herein incorporated in their entirety); and (4)
receptor-mediated mechanisms (Curiel et al., Hum. Gen. Ther.
3:147-154 (1992), Wagner et al., Proc. Natl. Acad. Sci. (USA)
89:6099-6103 (1992), both of which are incorporated by reference in
their entirety).
[0257] Acceleration methods that may be used include, for example,
microprojectile bombardment and the like. One example of a method
for delivering transforming nucleic acid molecules to plant cells
is microprojectile bombardment. This method has been reviewed by
Yang and Christou (eds.), Particle Bombardment Technology for Gene
Transfer, Oxford Press, Oxford, England (1994), the entirety of
which is herein incorporated by reference). Non-biological
particles (microprojectiles) that may be coated with nucleic acids
and delivered into cells by a propelling force. Exemplary particles
include those comprised of tungsten, gold, platinum and the
like.
[0258] A particular advantage of microprojectile bombardment, in
addition to it being an effective means of reproducibly
transforming monocots, is that neither the isolation of protoplasts
(Cristou et al., Plant Physiol. 87:671-674 (1988), the entirety of
which is herein incorporated by reference) nor the susceptibility
of Agrobacterium infection are required. An illustrative embodiment
of a method for delivering DNA into maize cells by acceleration is
a biolistics .alpha.-particle delivery system, which can be used to
propel particles coated with DNA through a screen, such as a
stainless steel or Nytex screen, onto a filter surface covered with
corn cells cultured in suspension. Gordon-Kamm et al., describes
the basic procedure for coating tungsten particles with DNA
(Gordon-Kamm et al., Plant Cell 2:603-618 (1990), the entirety of
which is herein incorporated by reference). The screen disperses
the tungsten nucleic acid particles so that they are not delivered
to the recipient cells in large aggregates. A particle delivery
system suitable for use with the present invention is the helium
acceleration PDS-1000/He gun is available from Bio-Rad Laboratories
(Bio-Rad, Hercules, Calif.)(Sanford et al., Technique 3:3-16
(1991), the entirety of which is herein incorporated by
reference).
[0259] For the bombardment, cells in suspension may be concentrated
on filters. Filters containing the cells to be bombarded are
positioned at an appropriate distance below the microprojectile
stopping plate. If desired, one or more screens are also positioned
between the gun and the cells to be bombarded.
[0260] Alternatively, immature embryos or other target cells may be
arranged on solid culture medium. The cells to be bombarded are
positioned at an appropriate distance below the microprojectile
stopping plate. If desired, one or more screens are also positioned
between the acceleration device and the cells to be bombarded.
Through the use of techniques set forth herein one may obtain up to
1000 or more foci of cells transiently expressing a marker gene.
The number of cells in a focus which express the exogenous gene
product 48 hours post-bombardment often range from one to ten and
average one to three.
[0261] In bombardment transformation, one may optimize the
pre-bombardment culturing conditions and the bombardment parameters
to yield the maximum numbers of stable transformants. Both the
physical and biological parameters for bombardment are important in
this technology. Physical factors are those that involve
manipulating the DNA/microprojectile precipitate or those that
affect the flight and velocity of either the macro- or
microprojectiles. Biological factors include all steps involved in
manipulation of cells before and immediately after bombardment, the
osmotic adjustment of target cells to help alleviate the trauma
associated with bombardment and also the nature of the transforming
DNA, such as linearized DNA or intact supercoiled plasmids. It is
believed that pre-bombardment manipulations are especially
important for successful transformation of immature embryos.
[0262] In another alternative embodiment, plastids can be stably
transformed. Methods disclosed for plastid transformation in higher
plants include the particle gun delivery of DNA containing a
selectable marker and targeting of the DNA to the plastid genome
through homologous recombination (Svab et al., Proc. Natl. Acad.
Sci. (U.S.A.) 87:8526-8530 (1990); Svab and Maliga, Proc. Natl.
Acad. Sci. (U.S.A.) 90:913-917 (1993); Staub and Maliga, EMBO J.
12:601-606 (1993); U.S. Pat. Nos. 5,451,513 and 5,545,818, all of
which are herein incorporated by reference in their entirety).
[0263] Accordingly, it is contemplated that one may wish to adjust
various aspects of the bombardment parameters in small scale
studies to fully optimize the conditions. One may particularly wish
to adjust physical parameters such as gap distance, flight
distance, tissue distance and helium pressure. One may also
minimize the trauma reduction factors by modifying conditions which
influence the physiological state of the recipient cells and which
may therefore influence transformation and integration
efficiencies. For example, the osmotic state, tissue hydration and
the subculture stage or cell cycle of the recipient cells may be
adjusted for optimum transformation. The execution of other routine
adjustments will be known to those of skill in the art in light of
the present disclosure.
[0264] Agrobacterium-mediated transfer is a widely applicable
system for introducing genes into plant cells because the DNA can
be introduced into whole plant tissues, thereby bypassing the need
for regeneration of an intact plant from a protoplast. The use of
Agrobacterium-mediated plant integrating vectors to introduce DNA
into plant cells is well known in the art. See, for example the
methods described by Fraley et al., Bio/Technology 3:629-635 (1985)
and Rogers et al., Methods Enzymol. 153:253-277 (1987), both of
which are herein incorporated by reference in their entirety.
Further, the integration of the Ti-DNA is a relatively precise
process resulting in few rearrangements. The region of DNA to be
transferred is defined by the border sequences and intervening DNA
is usually inserted into the plant genome as described (Spielmann
et al., Mol. Gen. Genet. 205:34 (1986), the entirety of which is
herein incorporated by reference).
[0265] Modern Agrobacterium transformation vectors are capable of
replication in E. coli as well as Agrobacterium, allowing for
convenient manipulations as described (Klee et al., In: Plant DNA
Infectious Agents, Hohn and Schell (eds.), Springer-Verlag, New
York, pp. 179-203 (1985), the entirety of which is herein
incorporated by reference. Moreover, technological advances in
vectors for Agrobacterium-mediated gene transfer have improved the
arrangement of genes and restriction sites in the vectors to
facilitate construction of vectors capable of expressing various
polypeptide coding genes. The vectors described have convenient
multi-linker regions flanked by a promoter and a polyadenylation
site for direct expression of inserted polypeptide coding genes and
are suitable for present purposes (Rogers et al., Methods Enzymol.
153:253-277 (1987)). In addition, Agrobacterium containing both
armed and disarmed Ti genes can be used for the transformations. In
those plant strains where Agrobacterium-mediated transformation is
efficient, it is the method of choice because of the facile and
defined nature of the gene transfer.
[0266] A transgenic plant formed using Agrobacterium transformation
methods typically contains a single gene on one chromosome. Such
transgenic plants can be referred to as being heterozygous for the
added gene. More preferred is a transgenic plant that is homozygous
for the added structural gene; i.e., a transgenic plant that
contains two added genes, one gene at the same locus on each
chromosome of a chromosome pair. A homozygous transgenic plant can
be obtained by sexually mating (selfing) an independent segregant
transgenic plant that contains a single added gene, germinating
some of the seed produced and analyzing the resulting plants
produced for the gene of interest.
[0267] It is also to be understood that two different transgenic
plants can also be mated to produce offspring that contain two
independently segregating added, exogenous genes. Selfing of
appropriate progeny can produce plants that are homozygous for both
added, exogenous genes that encode a polypeptide of interest.
Back-crossing to a parental plant and out-crossing with a
non-transgenic plant are also contemplated, as is vegetative
propagation.
[0268] Transformation of plant protoplasts can be achieved using
methods based on calcium phosphate precipitation, polyethylene
glycol treatment, electroporation and combinations of these
treatments (See, for example, Potrykus et al., Mol. Gen. Genet.
205:193-200 (1986); Lorz et al., Mol. Gen. Genet. 199:178 (1985);
Fromm et al., Nature 319:791 (1986); Uchimiya et al., Mol. Gen.
Genet. 204:204 (1986); Marcotte et al., Nature 335:454-457 (1988),
all of which are herein incorporated by reference in their
entirety).
[0269] Application of these systems to different plant strains
depends upon the ability to regenerate that particular plant strain
from protoplasts. Illustrative methods for the regeneration of
cereals from protoplasts are described (Fujimura et al., Plant
Tissue Culture Letters 2:74 (1985); Toriyama et al., Theor Appl.
Genet. 205:34 (1986); Yamada et al., Plant Cell Rep. 4:85 (1986);
Abdullah et al., Biotechnolog 4:1087 (1986), all of which are
herein incorporated by reference in their entirety).
[0270] To transform plant strains that cannot be successfully
regenerated from protoplasts, other ways to introduce DNA into
intact cells or tissues can be utilized. For example, regeneration
of cereals from immature embryos or explants can be effected as
described (Vasil, Biotechnology 6:397 (1988), the entirety of which
is herein incorporated by reference). In addition, "particle gun"
or high-velocity microprojectile technology can be utilized (Vasil
et al., Bio/Technology 10:667 (1992), the entirety of which is
herein incorporated by reference).
[0271] Using the latter technology, DNA is carried through the cell
wall and into the cytoplasm on the surface of small metal particles
as described (Klein et al., Nature 328:70 (1987); Klein et al.,
Proc. Natl. Acad. Sci. (U.S.A.) 85:8502-8505 (1988); McCabe et al.,
Bio/Technology 6:923 (1988), all of which are herein incorporated
by reference in their entirety). The metal particles penetrate
through several layers of cells and thus allow the transformation
of cells within tissue explants.
[0272] Other methods of cell transformation can also be used and
include but are not limited to introduction of DNA into plants by
direct DNA transfer into pollen (Zhou et al., Methods Enzymol.
101:433 (1983); Hess et al., Intern Rev. Cytol. 107:367 (1987); Luo
et al., Plant Mol. Biol. Reporter 6:165 (1988), all of which are
herein incorporated by reference in their entirety), by direct
injection of DNA into reproductive organs of a plant (Pena et al.,
Nature 325:274 (1987), the entirety of which is herein incorporated
by reference), or by direct injection of DNA into the cells of
immature embryos followed by the rehydration of desiccated embryos
(Neuhaus et al., Theor. Appl. Genet. 75:30 (1987), the entirety of
which is herein incorporated by reference).
[0273] The regeneration, development and cultivation of plants from
single plant protoplast transformants or from various transformed
explants is well known in the art (Weissbach and Weissbach, In:
Methods for Plant Molecular Biology, Academic Press, San Diego,
Calif., (1988), the entirety of which is herein incorporated by
reference). This regeneration and growth process typically includes
the steps of selection of transformed cells, culturing those
individualized cells through the usual stages of embryonic
development through the rooted plantlet stage. Transgenic embryos
and seeds are similarly regenerated. The resulting transgenic
rooted shoots are thereafter planted in an appropriate plant growth
medium such as soil.
[0274] The development or regeneration of plants containing the
foreign, exogenous gene that encodes a protein of interest is well
known in the art. Preferably, the regenerated plants are
self-pollinated to provide homozygous transgenic plants. Otherwise,
pollen obtained from the regenerated plants is crossed to
seed-grown plants of agronomically important lines. Conversely,
pollen from plants of these important lines is used to pollinate
regenerated plants. A transgenic plant of the present invention
containing a desired polypeptide is cultivated using methods well
known to one skilled in the art.
[0275] There are a variety of methods for the regeneration of
plants from plant tissue. The particular method of regeneration
will depend on the starting plant tissue and the particular plant
species to be regenerated.
[0276] Methods for transforming dicots, primarily by use of
Agrobacterium tumefaciens and obtaining transgenic plants have been
published for cotton (U.S. Pat. No. 5,004,863; U.S. Pat. No.
5,159,135; U.S. Pat. No. 5,518,908, all of which are herein
incorporated by reference in their entirety); soybean (U.S. Pat.
No. 5,569,834; U.S. Pat. No. 5,416,011; McCabe et. al.,
Biotechnology 6:923 (1988); Christou et al., Plant Physiol.
87:671-674 (1988); all of which are herein incorporated by
reference in their entirety); Brassica (U.S. Pat. No. 5,463,174,
the entirety of which is herein incorporated by reference); peanut
(Cheng et al., Plant Cell Rep. 15:653-657 (1996), McKently et al.,
Plant Cell Rep. 14:699-703 (1995), all of which are herein
incorporated by reference in their entirety); papaya; and pea
(Grant et al., Plant Cell Rep. 15:254-258 (1995), the entirety of
which is herein incorporated by reference).
[0277] Transformation of monocotyledons using electroporation,
particle bombardment and Agrobacterium have also been reported.
Transformation and plant regeneration have been achieved in
asparagus (Bytebier et al., Proc. Natl. Acad. Sci. (USA) 84:5354
(1987), the entirety of which is herein incorporated by reference);
barley (Wan and Lemaux, Plant Physiol 104:37 (1994), the entirety
of which is herein incorporated by reference); maize (Rhodes et
al., Science 240:204 (1988); Gordon-Kamm et al., Plant Cell
2:603-618 (1990); Fromm et al., Bio/Technology 8:833 (1990); Koziel
et al., Bio/Technology 11:194 (1993); Armstrong et al., Crop
Science 35:550-557 (1995); all of which are herein incorporated by
reference in their entirety); oat (Somers et al., Bio/Technology
10:1589 (1992), the entirety of which is herein incorporated by
reference); orchard grass (Horn et al., Plant Cell Rep. 7:469
(1988), the entirety of which is herein incorporated by reference);
rice (Toriyama et al., Theor Appl. Genet. 205:34 (1986); Part et
al., Plant Mol. Biol. 32:1135-1148 (1996); Abedinia et al., Aust.
J. Plant Physiol. 24:133-141 (1997); Zhang and Wu, Theor. Appl.
Genet. 76:835 (1988); Zhang et al., Plant Cell Rep. 7:379 (1988);
Battraw and Hall, Plant Sci. 86:191-202 (1992); Christou et al.,
Bio/Technology 9:957 (1991), all of which are herein incorporated
by reference in their entirety); rye (De la Pena et al., Nature
325:274 (1987), the entirety of which is herein incorporated by
reference); sugarcane (Bower and Birch, Plant J. 2:409 (1992), the
entirety of which is herein incorporated by reference); tall fescue
(Wang et al., Bio/Technology 10:691 (1992), the entirety of which
is herein incorporated by reference) and wheat (Vasil et al.,
Bio/Technology 10:667 (1992), the entirety of which is herein
incorporated by reference; U.S. Pat. No. 5,631,152, the entirety of
which is herein incorporated by reference.)
[0278] Assays for gene expression based on the transient expression
of cloned nucleic acid constructs have been developed by
introducing the nucleic acid molecules into plant cells by
polyethylene glycol treatment, electroporation, or particle
bombardment (Marcotte et al., Nature 335:454-457 (1988), the
entirety of which is herein incorporated by reference; Marcotte et
al., Plant Cell 1:523-532 (1989), the entirety of which is herein
incorporated by reference; McCarty et al., Cell 66:895-905 (1991),
the entirety of which is herein incorporated by reference; Hattori
et al., Genes Dev. 6:609-618 (1992), the entirety of which is
herein incorporated by reference; Goff et al., EMBO J. 9:2517-2522
(1990), the entirety of which is herein incorporated by reference).
Transient expression systems may be used to functionally dissect
gene constructs (see generally, Mailga et al., Methods in Plant
Molecular Biology, Cold Spring Harbor Press (1995)).
[0279] Any of the nucleic acid molecules of the present invention
may be introduced into a plant cell in a permanent or transient
manner in combination with other genetic elements such as vectors,
promoters, enhancers etc. Further, any of the nucleic acid
molecules of the present invention may be introduced into a plant
cell in a manner that allows for overexpression of the protein or
fragment thereof encoded by the nucleic acid molecule.
[0280] Cosuppression is the reduction in expression levels, usually
at the level of RNA, of a particular endogenous gene or gene family
by the expression of a homologous sense construct that is capable
of transcribing mRNA of the same strandedness as the transcript of
the endogenous gene (Napoli et al., Plant Cell 2:279-289 (1990),
the entirety of which is herein incorporated by reference; van der
Krol et al., Plant Cell 2:291-299 (1990), the entirety of which is
herein incorporated by reference). Cosuppression may result from
stable transformation with a single copy nucleic acid molecule that
is homologous to a nucleic acid sequence found with the cell
(Prolls and Meyer, Plant J. 2:465-475 (1992), the entirety of which
is herein incorporated by reference) or with multiple copies of a
nucleic acid molecule that is homologous to a nucleic acid sequence
found with the cell (Mittlesten et al., Mot Gen. Genet. 244:325-330
(1994), the entirety of which is herein incorporated by reference).
Genes, even though different, linked to homologous promoters may
result in the cosuppression of the linked genes (Vaucheret, C. R.
Acad. Sci. III 316:1471-1483 (1993), the entirety of which is
herein incorporated by reference).
[0281] This technique has, for example, been applied to generate
white flowers from red petunia and tomatoes that do not ripen on
the vine. Up to 50% of petunia transformants that contained a sense
copy of the glucoamylase (CHS) gene produced white flowers or
floral sectors; this was as a result of the post-transcriptional
loss of mRNA encoding CHS (Flavell, Proc. Natl. Acad. Sci. (U.S.A.)
91:3490-3496 (1994), the entirety of which is herein incorporated
by reference); van Blokland et al., Plant J. 6:861-877 (1994), the
entirety of which is herein incorporated by reference).
Cosuppression may require the coordinate transcription of the
transgene and the endogenous gene and can be reset by a
developmental control mechanism (Jorgensen, Trends Biotechnol.
8:340-344 (1990), the entirety of which is herein incorporated by
reference; Meins and Kunz, In: Gene Inactivation and Homologous
Recombination in Plants, Paszkowski (ed.), pp. 335-348, Kluwer
Academic, Netherlands (1994), the entirety of which is herein
incorporated by reference).
[0282] It is understood that one or more of the nucleic acids of
the present invention may be introduced into a plant cell and
transcribed using an appropriate promoter with such transcription
resulting in the cosuppression of an endogenous methionine pathway
protein.
[0283] Antisense approaches are a way of preventing or reducing
gene function by targeting the genetic material (Mol et al., FEBS
Lett. 268:427-430 (1990), the entirety of which is herein
incorporated by reference). The objective of the antisense approach
is to use a sequence complementary to the target gene to block its
expression and create a mutant cell line or organism in which the
level of a single chosen protein is selectively reduced or
abolished. Antisense techniques have several advantages over other
`reverse genetic` approaches. The site of inactivation and its
developmental effect can be manipulated by the choice of promoter
for antisense genes or by the timing of external application or
microinjection. Antisense can manipulate its specificity by
selecting either unique regions of the target gene or regions where
it shares homology to other related genes (Hiatt et al., In:
Genetic Engineering, Setlow (ed.), Vol. 11, New York: Plenum 49-63
(1989), the entirety of which is herein incorporated by
reference).
[0284] The principle of regulation by antisense RNA is that RNA
that is complementary to the target mRNA is introduced into cells,
resulting in specific RNA:RNA duplexes being formed by base pairing
between the antisense substrate and the target mRNA (Green et al.,
Annu. Rev. Biochem. 55:569-597 (1986), the entirety of which is
herein incorporated by reference). Under one embodiment, the
process involves the introduction and expression of an antisense
gene sequence. Such a sequence is one in which part or all of the
normal gene sequences are placed under a promoter in inverted
orientation so that the `wrong` or complementary strand is
transcribed into a noncoding antisense RNA that hybridizes with the
target mRNA and interferes with its expression (Takayama and
Inouye, Crit. Rev. Biochem. Mol. Biol. 25:155-184 (1990), the
entirety of which is herein incorporated by reference). An
antisense vector is constructed by standard procedures and
introduced into cells by transformation, transfection,
electroporation, microinjection, infection, etc. The type of
transformation and choice of vector will determine whether
expression is transient or stable. The promoter used for the
antisense gene may influence the level, timing, tissue,
specificity, or inducibility of the antisense inhibition.
[0285] It is understood that the activity of a methionine pathway
protein in a plant cell may be reduced or depressed by growing a
transformed plant cell containing a nucleic acid molecule whose
non-transcribed strand encodes a methionine pathway protein or
fragment thereof.
[0286] Antibodies have been expressed in plants (Hiatt et al.,
Nature 342:76-78 (1989), the entirety of which is herein
incorporated by reference; Conrad and Fielder, Plant Mol. Biol.
26:1023-1030 (1994), the entirety of which is herein incorporated
by reference). Cytoplamsic expression of a scFv (single-chain Fv
antibodies) has been reported to delay infection by artichoke
mottled crinkle virus. Transgenic plants that express antibodies
directed against endogenous proteins may exhibit a physiological
effect (Philips et al., EMBO J. 16:4489-4496 (1997), the entirety
of which is herein incorporated by reference; Marion-Poll, Trends
in Plant Science 2:447-448 (1997), the entirety of which is herein
incorporated by reference). For example, expressed anti-abscisic
antibodies have been reported to result in a general perturbation
of seed development (Philips et al., EMBO J. 16: 4489-4496
(1997)).
[0287] Antibodies that are catalytic may also be expressed in
plants (abzymes). The principle behind abzymes is that since
antibodies may be raised against many molecules, this recognition
ability can be directed toward generating antibodies that bind
transition states to force a chemical reaction forward (Persidas,
Nature Biotechnology 15:1313-1315 (1997), the entirety of which is
herein incorporated by reference; Baca et al., Ann. Rev. Biophys.
Biomol. Struct. 26:461-493 (1997), the entirety of which is herein
incorporated by reference). The catalytic abilities of abzymes may
be enhanced by site directed mutagenesis. Examples of abzymes are,
for example, set forth in U.S. Pat. No. 5,658,753; U.S. Pat. No.
5,632,990; U.S. Pat. No. 5,631,137; U.S. Pat. No. 5,602,015; U.S.
Pat. No. 5,559,538; U.S. Pat. No. 5,576,174; U.S. Pat. No.
5,500,358; U.S. Pat. No. 5,318,897; U.S. Pat. No. 5,298,409; U.S.
Pat. No. 5,258,289 and U.S. Pat. No. 5,194,585, all of which are
herein incorporated in their entirety.
[0288] It is understood that any of the antibodies of the present
invention may be expressed in plants and that such expression can
result in a physiological effect. It is also understood that any of
the expressed antibodies may be catalytic.
[0289] (b) Fungal Constructs and Fungal Transformants
[0290] The present invention also relates to a fungal recombinant
vector comprising exogenous genetic material. The present invention
also relates to a fungal cell comprising a fungal recombinant
vector. The present invention also relates to methods for obtaining
a recombinant fungal host cell comprising introducing into a fungal
host cell exogenous genetic material.
[0291] Exogenous genetic material may be transferred into a fungal
cell. In a preferred embodiment the exogenous genetic material
includes a nucleic acid molecule of the present invention having a
sequence selected from the group consisting of SEQ ID NO: 1 through
SEQ ID NO: 3204 or complements thereof or fragments of either. The
fungal recombinant vector may be any vector which can be
conveniently subjected to recombinant DNA procedures. The choice of
a vector will typically depend on the compatibility of the vector
with the fungal host cell into which the vector is to be
introduced. The vector may be a linear or a closed circular
plasmid. The vector system may be a single vector or plasmid or two
or more vectors or plasmids which together contain the total DNA to
be introduced into the genome of the fungal host.
[0292] The fungal vector may be an autonomously replicating vector,
i.e., a vector which exists as an extrachromosomal entity, the
replication of which is independent of chromosomal replication,
e.g., a plasmid, an extrachromosomal element, a minichromosome, or
an artificial chromosome. The vector may contain any means for
assuring self-replication. Alternatively, the vector may be one
which, when introduced into the fungal cell, is integrated into the
genome and replicated together with the chromosome(s) into which it
has been integrated. For integration, the vector may rely on the
nucleic acid sequence of the vector for stable integration of the
vector into the genome by homologous or nonhomologous
recombination. Alternatively, the vector may contain additional
nucleic acid sequences for directing integration by homologous
recombination into the genome of the fungal host. The additional
nucleic acid sequences enable the vector to be integrated into the
host cell genome at a precise location(s) in the chromosome(s). To
increase the likelihood of integration at a precise location, there
should be preferably two nucleic acid sequences which individually
contain a sufficient number of nucleic acids, preferably 400 bp to
1500 bp, more preferably 800 bp to 1000 bp, which are highly
homologous with the corresponding target sequence to enhance the
probability of homologous recombination. These nucleic acid
sequences may be any sequence that is homologous with a target
sequence in the genome of the fungal host cell and, furthermore,
may be non-encoding or encoding sequences.
[0293] For autonomous replication, the vector may further comprise
an origin of replication enabling the vector to replicate
autonomously in the host cell in question. Examples of origin of
replications for use in a yeast host cell are the 2 micron origin
of replication and the combination of CEN3 and ARS1. Any origin of
replication may be used which is compatible with the fungal host
cell of choice.
[0294] The fungal vectors of the present invention preferably
contain one or more selectable markers which permit easy selection
of transformed cells. A selectable marker is a gene the product of
which provides, for example biocide or viral resistance, resistance
to heavy metals, prototrophy to auxotrophs and the like. The
selectable marker may be selected from the group including, but not
limited to, amdS (acetamidase), argB (ornithine
carbamoyltransferase), bar (phosphinothricin acetyltransferase),
hygB (hygromycin phosphotransferase), niaD (nitrate reductase),
pyrG (orotidine-5'-phosphate decarboxylase) and sC (sulfate
adenyltransferase) and trpC (anthranilate synthase). Preferred for
use in an Aspergillus cell are the amdS and pyrG markers of
Aspergillus nidulans or Aspergillus oryzae and the bar marker of
Streptomyces hygroscopicus. Furthermore, selection may be
accomplished by co-transformation, e.g., as described in WO
91/17243, the entirety of which is herein incorporated by
reference. A nucleic acid sequence of the present invention may be
operably linked to a suitable promoter sequence. The promoter
sequence is a nucleic acid sequence which is recognized by the
fungal host cell for expression of the nucleic acid sequence. The
promoter sequence contains transcription and translation control
sequences which mediate the expression of the protein or fragment
thereof.
[0295] A promoter may be any nucleic acid sequence which shows
transcriptional activity in the fungal host cell of choice and may
be obtained from genes encoding polypeptides either homologous or
heterologous to the host cell. Examples of suitable promoters for
directing the transcription of a nucleic acid construct of the
invention in a filamentous fungal host are promoters obtained from
the genes encoding Aspergillus oryzae TAKA amylase, Rhizomucor
miehei aspartic proteinase, Aspergillus niger neutral
alpha-amylase, Aspergillus niger acid stable alpha-amylase,
Aspergillus niger or Aspergillus awamori glucoamylase (glaA),
Rhizomucor miehei lipase, Aspergillus oryzae alkaline protease,
Aspergillus oryzae triose phosphate isomerase, Aspergillus nidulans
acetamidase and hybrids thereof. In a yeast host, a useful promoter
is the Saccharomyces cerevisiae enolase (eno-1) promoter.
Particularly preferred promoters are the TAKA amylase, NA2-tpi (a
hybrid of the promoters from the genes encoding Aspergillus niger
neutral alpha-amylase and Aspergillus oryzae triose phosphate
isomerase) and glaA promoters.
[0296] A protein or fragment thereof encoding nucleic acid molecule
of the present invention may also be operably linked to a
terminator sequence at its 3' terminus. The terminator sequence may
be native to the nucleic acid sequence encoding the protein or
fragment thereof or may be obtained from foreign sources. Any
terminator which is functional in the fungal host cell of choice
may be used in the present invention, but particularly preferred
terminators are obtained from the genes encoding Aspergillus oryzae
TAKA amylase, Aspergillus niger glucoamylase, Aspergillus nidulans
anthranilate synthase, Aspergillus niger alpha-glucosidase and
Saccharomyces cerevisiae enolase.
[0297] A protein or fragment thereof encoding nucleic acid molecule
of the present invention may also be operably linked to a suitable
leader sequence. A leader sequence is a nontranslated region of a
mRNA which is important for translation by the fungal host. The
leader sequence is operably linked to the 5' terminus of the
nucleic acid sequence encoding the protein or fragment thereof. The
leader sequence may be native to the nucleic acid sequence encoding
the protein or fragment thereof or may be obtained from foreign
sources. Any leader sequence which is functional in the fungal host
cell of choice may be used in the present invention, but
particularly preferred leaders are obtained from the genes encoding
Aspergillus oryzae TAKA amylase and Aspergillus oryzae triose
phosphate isomerase.
[0298] A polyadenylation sequence may also be operably linked to
the 3' terminus of the nucleic acid sequence of the present
invention. The polyadenylation sequence is a sequence which when
transcribed is recognized by the fungal host to add polyadenosine
residues to transcribed mRNA. The polyadenylation sequence may be
native to the nucleic acid sequence encoding the protein or
fragment thereof or may be obtained from foreign sources. Any
polyadenylation sequence which is functional in the fungal host of
choice may be used in the present invention, but particularly
preferred polyadenylation sequences are obtained from the genes
encoding Aspergillus oryzae TAKA amylase, Aspergillus niger
glucoamylase, Aspergillus nidulans anthranilate synthase and
Aspergillus niger alpha-glucosidase.
[0299] To avoid the necessity of disrupting the cell to obtain the
protein or fragment thereof and to minimize the amount of possible
degradation of the expressed protein or fragment thereof within the
cell, it is preferred that expression of the protein or fragment
thereof gives rise to a product secreted outside the cell. To this
end, a protein or fragment thereof of the present invention may be
linked to a signal peptide linked to the amino terminus of the
protein or fragment thereof. A signal peptide is an amino acid
sequence which permits the secretion of the protein or fragment
thereof from the fungal host into the culture medium. The signal
peptide may be native to the protein or fragment thereof of the
invention or may be obtained from foreign sources. The 5' end of
the coding sequence of the nucleic acid sequence of the present
invention may inherently contain a signal peptide coding region
naturally linked in translation reading frame with the segment of
the coding region which encodes the secreted protein or fragment
thereof. Alternatively, the 5' end of the coding sequence may
contain a signal peptide coding region which is foreign to that
portion of the coding sequence which encodes the secreted protein
or fragment thereof. The foreign signal peptide may be required
where the coding sequence does not normally contain a signal
peptide coding region. Alternatively, the foreign signal peptide
may simply replace the natural signal peptide to obtain enhanced
secretion of the desired protein or fragment thereof. The foreign
signal peptide coding region may be obtained from a glucoamylase or
an amylase gene from an Aspergillus species, a lipase or proteinase
gene from Rhizomucor miehei, the gene for the alpha-factor from
Saccharomyces cerevisiae, or the calf preprochymosin gene. An
effective signal peptide for fungal host cells is the Aspergillus
oryzae TAKA amylase signal, Aspergillus niger neutral amylase
signal, the Rhizomucor miehei aspartic proteinase signal, the
Humicola lanuginosus cellulase signal, or the Rhizomucor miehei
lipase signal. However, any signal peptide capable of permitting
secretion of the protein or fragment thereof in a fungal host of
choice may be used in the present invention.
[0300] A protein or fragment thereof encoding nucleic acid molecule
of the present invention may also be linked to a propeptide coding
region. A propeptide is an amino acid sequence found at the amino
terminus of aproprotein or proenzyme. Cleavage of the propeptide
from the proprotein yields a mature biochemically active protein.
The resulting polypeptide is known as a propolypeptide or proenzyme
(or a zymogen in some cases). Propolypeptides are generally
inactive and can be converted to mature active polypeptides by
catalytic or autocatalytic cleavage of the propeptide from the
propolypeptide or proenzyme. The propeptide coding region may be
native to the protein or fragment thereof or may be obtained from
foreign sources. The foreign propeptide coding region may be
obtained from the Saccharomyces cerevisiae alpha-factor gene or
Myceliophthora thermophila laccase gene (WO 95/33836, the entirety
of which is herein incorporated by reference).
[0301] The procedures used to ligate the elements described above
to construct the recombinant expression vector of the present
invention are well known to one skilled in the art (see, for
example, Sambrook et al., Molecular Cloning, A Laboratory Manual,
2nd ed., Cold Spring Harbor, N.Y., (1989)).
[0302] The present invention also relates to recombinant fungal
host cells produced by the methods of the present invention which
are advantageously used with the recombinant vector of the present
invention. The cell is preferably transformed with a vector
comprising a nucleic acid sequence of the invention followed by
integration of the vector into the host chromosome. The choice of
fungal host cells will to a large extent depend upon the gene
encoding the protein or fragment thereof and its source. The fungal
host cell may, for example, be a yeast cell or a filamentous fungal
cell.
[0303] "Yeast" as used herein includes Ascosporogenous yeast
(Endomycetales), Basidiosporogenous yeast and yeast belonging to
the Fungi Imperfecti (Blastomycetes). The Ascosporogenous yeasts
are divided into the families Spermophthoraceae and
Saccharomycetaceae. The latter is comprised of four subfamilies,
Schizosaccharomycoideae (for example, genus Schizosaccharomyces),
Nadsonioideae, Lipomycoideae and Saccharomycoideae (for example,
genera Pichia, Kluyveromyces and Saccharomyces). The
Basidiosporogenous yeasts include the genera Leucosporidim,
Rhodosporidium, Sporidiobolus, Filobasidium and Filobasidiella.
Yeast belonging to the Fungi Imperfecti are divided into two
families, Sporobolomycetaceae (for example, genera Sorobolomyces
and Bullera) and Cryptococcaceae (for example, genus Candida).
Since the classification of yeast may change in the future, for the
purposes of this invention, yeast shall be defined as described in
Biology and Activities of Yeast (Skinner et al., Soc. App.
Bacterial. Symposium Series No. 9, (1980), the entirety of which is
herein incorporated by reference). The biology of yeast and
manipulation of yeast genetics are well known in the art (see, for
example, Biochemistry and Genetics of Yeast, Bacil et al. (ed.),
2nd edition, 1987; The Yeasts, Rose and Harrison (eds.), 2nd ed.,
(1987); and The Molecular Biology of the Yeast Saccharomyces,
Strathern et al. (eds.), (1981), all of which are herein
incorporated by reference in their entirety).
[0304] "Fungi" as used herein includes the phyla Ascomycota,
Basidiomycota, Chytridiomycota and Zygomycota (as defined by
Hawksworth et al., In: Ainsworth and Bisby's Dictionary of The
Fungi, 8th edition, 1995, CAB International, University Press,
Cambridge, UK; the entirety of which is herein incorporated by
reference) as well as the Oomycota (as cited in Hawksworth et al.,
In: Ainsworth and Bisby's Dictionary of The Fungi, 8th edition,
1995, CAB International, University Press, Cambridge, UK) and all
mitosporic fungi (Hawksworth et al., In: Ainsworth and Bisby's
Dictionary of The Fungi, 8th edition, 1995, CAB International,
University Press, Cambridge, UK). Representative groups of
Ascomycota include, for example, Neurospora, Eupenicillium
(=Penicillium), Emericella (=Aspergillus), Eurotiun (=Aspergillus)
and the true yeasts listed above. Examples of Basidiomycota include
mushrooms, rusts and smuts. Representative groups of
Chytridiomycota include, for example, Allomyces, Blastocladiella,
Coelomomyces and aquatic fungi. Representative groups of Oomycota
include, for example, Saprolegniomycetous aquatic fungi (water
molds) such as Achlya. Examples of mitosporic fungi include
Aspergillus, Penicilliun, Candida and Alternaria. Representative
groups of Zygomycota include, for example, Rhizopus and Mucor.
[0305] "Filamentous fungi" include all filamentous forms of the
subdivision Eumycota and Oomycota (as defined by Hawksworth et al.,
In: Ainsworth and Bisby's Dictionary of The Fungi, 8th edition,
1995, CAB International, University Press, Cambridge, UK). The
filamentous fungi are characterized by a vegetative mycelium
composed of chitin, cellulose, glucan, chitosan, mannan and other
complex polysaccharides. Vegetative growth is by hyphal elongation
and carbon catabolism is obligately aerobic. In contrast,
vegetative growth by yeasts such as Saccharomyces cerevisiae is by
budding of a unicellular thallus and carbon catabolism may be
fermentative.
[0306] In one embodiment, the fungal host cell is a yeast cell. In
a preferred embodiment, the yeast host cell is a cell of the
species of Candida, Kluyveromyces, Saccharomyces,
Schizosaccharomyces, Pichia and Yarrowia. In a preferred
embodiment, the yeast host cell is a Saccharomyces cerevisiae cell,
a Saccharomyces carlsbergensis, Saccharomyces diastaticus cell, a
Saccharomyces douglasii cell, a Saccharomyces kluyveri cell, a
Saccharomyces norbensis cell, or a Saccharomyces oviformis cell. In
another preferred embodiment, the yeast host cell is a
Kluyveromyces lactis cell. In another preferred embodiment, the
yeast host cell is a Yarrowia lipolytica cell.
[0307] In another embodiment, the fungal host cell is a filamentous
fungal cell. In a preferred embodiment, the filamentous fungal host
cell is a cell of the species of, but not limited to, Acremonium,
Aspergillus, Fusarium, Humicola, Myceliophthora, Mucor, Neurospora,
Penicillium, Thielavia, Tolypocladium and Trichoderma. In a
preferred embodiment, the filamentous fungal host cell is an
Aspergillus cell. In another preferred embodiment, the filamentous
fungal host cell is an Acremonium cell. In another preferred
embodiment, the filamentous fungal host cell is a Fusarium cell. In
another preferred embodiment, the filamentous fungal host cell is a
Humicola cell. In another preferred embodiment, the filamentous
fungal host cell is a Myceliophthora cell. In another even
preferred embodiment, the filamentous fungal host cell is a Mucor
cell. In another preferred embodiment, the filamentous fungal host
cell is a Neurospora cell. In another preferred embodiment, the
filamentous fungal host cell is a Penicillium cell. In another
preferred embodiment, the filamentous fungal host cell is a
Thielavia cell. In another preferred embodiment, the filamentous
fungal host cell is a Tolypocladiun cell. In another preferred
embodiment, the filamentous fungal host cell is a Trichoderma cell.
In a preferred embodiment, the filamentous fungal host cell is an
Aspergillus oryzae cell, an Aspergillus niger cell, an Aspergillus
foetidus cell, or an Aspergillus japonicus cell. In another
preferred embodiment, the filamentous fungal host cell is a
Fusarium oxysporum cell or a Fusarium graminearum cell. In another
preferred embodiment, the filamentous fungal host cell is a
Humicola insolens cell or a Humicola lanuginosus cell. In another
preferred embodiment, the filamentous fungal host cell is a
Myceliophthora thermophila cell. In a most preferred embodiment,
the filamentous fungal host cell is a Mucor miehei cell. In a most
preferred embodiment, the filamentous fungal host cell is a
Neurospora crassa cell. In a most preferred embodiment, the
filamentous fungal host cell is a Penicillium purpurogenum cell. In
another most preferred embodiment, the filamentous fungal host cell
is a Thielavia terrestris cell. In another most preferred
embodiment, the Trichoderma cell is a Trichoderma reesei cell, a
Trichoderma viride cell, a Trichoderma longibrachiatum cell, a
Trichoderma harzianum cell, or a Trichoderma koningii cell. In a
preferred embodiment, the fungal host cell is selected from an A.
nidulans cell, an A. niger cell, an A. oryzae cell and an A. sojae
cell. In a further preferred embodiment, the fungal host cell is an
A. nidulans cell.
[0308] The recombinant fungal host cells of the present invention
may further comprise one or more sequences which encode one or more
factors that are advantageous in the expression of the protein or
fragment thereof, for example, an activator (e.g., a trans-acting
factor), a chaperone and a processing protease. The nucleic acids
encoding one or more of these factors are preferably not operably
linked to the nucleic acid encoding the protein or fragment
thereof. An activator is a protein which activates transcription of
a nucleic acid sequence encoding a polypeptide (Kudla et al., EMBO
9:1355-1364 (1990); Jarai and Buxton, Current Genetics 26:2238-244
(1994); Verdier, Yeast 6:271-297 (1990), all of which are herein
incorporated by reference in their entirety). The nucleic acid
sequence encoding an activator may be obtained from the genes
encoding Saccharomyces cerevisiae heme activator protein 1 (hapl),
Saccharomyces cerevisiae galactose metabolizing protein 4 (gal4)
and Aspergillus nidulans ammonia regulation protein (areA). For
further examples, see Verdier, Yeast 6:271-297 (1990); MacKenzie et
al., Journal of Gen. Microbiol. 139:2295-2307 (1993), both of which
are herein incorporated by reference in their entirety). A
chaperone is a protein which assists another protein in folding
properly (Hartl et al., TIBS 19:20-25 (1994); Bergeron et al.,
TIBS19:124-128 (1994); Demolder et al., 0.1 Biotechnology
32:179-189 (1994); Craig, Science 260:1902-1903 (1993); Gething and
Sambrook, Nature 355:33-45 (1992); Puig and Gilbert, J Biol. Chem.
269:7764-7771 (1994); Wang and Tsou, FASEB Journal 7:1515-11157
(1993); Robinson et al., Bio/Technology 1:381-384 (1994), all of
which are herein incorporated by reference in their entirety). The
nucleic acid sequence encoding a chaperone may be obtained from the
genes encoding Aspergillus oryzae protein disulphide isomerase,
Saccharomyces cerevisiae calnexin, Saccharomyces cerevisiae
BiP/GRP78 and Saccharomyces cerevisiae Hsp70. For further examples,
see Gething and Sambrook, Nature 355:33-45 (1992); Hartl et al.,
TIBS 19:20-25 (1994). A processing protease is a protease that
cleaves a propeptide to generate a mature biochemically active
polypeptide (Enderlin and Ogrydziak, Yeast 10:67-79 (1994); Fuller
et al., Proc. Natl. Acad. Sci. (U.S.A.) 86:1434-1438 (1989); Julius
et al., Cell 37:1075-1089 (1984); Julius et al., Cell 32:839-852
(1983), all of which are incorporated by reference in their
entirety). The nucleic acid sequence encoding a processing protease
may be obtained from the genes encoding Aspergillus niger Kex2,
Saccharomyces cerevisiae dipeptidylaminopeptidase, Saccharomyces
cerevisiae Kex2 and Yarrowia lipolytica dibasic processing
endoprotease (xpr6). Any factor that is functional in the fungal
host cell of choice may be used in the present invention.
[0309] Fungal cells may be transformed by a process involving
protoplast formation, transformation of the protoplasts and
regeneration of the cell wall in a manner known per se. Suitable
procedures for transformation of Aspergillus host cells are
described in EP 238 023 and Yelton et al., Proc. Natl. Acad. Sci.
(U.S.A.) 81:1470-1474 (1984), both of which are herein incorporated
by reference in their entirety. A suitable method of transforming
Fusarium species is described by Malardier et al., Gene 78:147-156
(1989), the entirety of which is herein incorporated by reference.
Yeast may be transformed using the procedures described by Becker
and Guarente, In: Abelson and Simon, (eds.), Guide to Yeast
Genetics and Molecular Biology, Methods Enzymol. Volume 194, pp
182-187, Academic Press, Inc., New York; Ito et al., J.
Bacteriology 153:163 (1983); Hinnen et al., Proc. Natl. Acad. Sci.
(U.S.A.) 75:1920 (1978), all of which are herein incorporated by
reference in their entirety.
[0310] The present invention also relates to methods of producing
the protein or fragment thereof comprising culturing the
recombinant fungal host cells under conditions conducive for
expression of the protein or fragment thereof. The fungal cells of
the present invention are cultivated in a nutrient medium suitable
for production of the protein or fragment thereof using methods
known in the art. For example, the cell may be cultivated by shake
flask cultivation, small-scale or large-scale fermentation
(including continuous, batch, fed-batch, or solid state
fermentations) in laboratory or industrial fermentors performed in
a suitable medium and under conditions allowing the protein or
fragment thereof to be expressed and/or isolated. The cultivation
takes place in a suitable nutrient medium comprising carbon and
nitrogen sources and inorganic salts, using procedures known in the
art (see, e.g., Bennett and LaSure (eds.), More Gene Manipulations
in Fungi, Academic Press, CA, (1991), the entirety of which is
herein incorporated by reference). Suitable media are available
from commercial suppliers or may be prepared according to published
compositions (e.g., in catalogues of the American Type Culture
Collection, Manassas, Va.). If the protein or fragment thereof is
secreted into the nutrient medium, a protein or fragment thereof
can be recovered directly from the medium. If the protein or
fragment thereof is not secreted, it is recovered from cell
lysates.
[0311] The expressed protein or fragment thereof may be detected
using methods known in the art that are specific for the particular
protein or fragment. These detection methods may include the use of
specific antibodies, formation of an enzyme product, or
disappearance of an enzyme) substrate. For example, if the protein
or fragment thereof has enzymatic activity, an enzyme assay may be
used. Alternatively, if polyclonal or monoclonal antibodies
specific to the protein or fragment thereof are available,
immunoassays may be employed using the antibodies to the protein or
fragment thereof. The techniques of enzyme assay and immunoassay
are well known to those skilled in the art.
[0312] The resulting protein or fragment thereof may be recovered
by methods known in the arts. For example, the protein or fragment
thereof may be recovered from the nutrient medium by conventional
procedures including, but not limited to, centrifugation,
filtration, extraction, spray-drying, evaporation, or
precipitation. The recovered protein or fragment thereof may then
be further purified by a variety of chromatographic procedures,
e.g., ion exchange chromatography, gel filtration chromatography,
affinity chromatography, or the like.
[0313] (c) Mammalian Constructs and Transformed Mammalian Cells
[0314] The present invention also relates to methods for obtaining
a recombinant mammalian host cell, comprising introducing into a
mammalian host cell exogenous genetic material. The present
invention also relates to a mammalian cell comprising a mammalian
recombinant vector. The present invention also relates to methods
for obtaining a recombinant mammalian host cell, comprising
introducing into a mammalian cell exogenous genetic material.
[0315] Mammalian cell lines available as hosts for expression are
known in the art and include many immortalized cell lines available
from the American Type Culture Collection (ATCC, Manassas, Va.),
such as HeLa cells, Chinese hamster ovary (CHO) cells, baby hamster
kidney (BHK) cells and a number of other cell lines. Suitable
promoters for mammalian cells are also known in the art and include
viral promoters such as that from Simian Virus 40 (SV40) (Fiers et
al., Nature 273:113 (1978), the entirety of which is herein
incorporated by reference), Rous sarcoma virus (RSV), adenovirus
(ADV) and bovine papilloma virus (BPV). Mammalian cells may also
require terminator sequences and poly-A addition sequences.
Enhancer sequences which increase expression may also be included
and sequences which promote amplification of the gene may also be
desirable (for example methotrexate resistance genes).
[0316] Vectors suitable for replication in mammalian cells may
include viral replicons, or sequences which insure integration of
the appropriate sequences encoding HCV epitopes into the host
genome. For example, another vector used to express foreign DNA is
vaccinia virus. In this case, for example, a nucleic acid molecule
encoding a protein or fragment thereof is inserted into the
vaccinia genome. Techniques for the insertion of foreign DNA into
the vaccinia virus genome are known in the art and may utilize, for
example, homologous recombination. Such heterologous DNA is
generally inserted into a gene which is non-essential to the virus,
for example, the thymidine kinase gene (tk), which also provides a
selectable marker. Plasmid vectors that greatly facilitate the
construction of recombinant viruses have been described (see, for
example, Mackett et al, J. Virol. 49:857 (1984); Chakrabarti et
al., Mol. Cell. Biol. 5:3403 (1985); Moss, In: Gene Transfer
Vectors For Mammalian Cells (Miller and Calos, eds., Cold Spring
Harbor Laboratory, N.Y., p. 10, (1987); all of which are herein
incorporated by reference in their entirety). Expression of the HCV
polypeptide then occurs in cells or animals which are infected with
the live recombinant vaccinia virus.
[0317] The sequence to be integrated into the mammalian sequence
may be introduced into the primary host by any convenient means,
which includes calcium precipitated DNA, spheroplast fusion,
transformation, electroporation, biolistics, lipofection,
microinjection, or other convenient means. Where an amplifiable
gene is being employed, the amplifiable gene may serve as the
selection marker for selecting hosts into which the amplifiable
gene has been introduced. Alternatively, one may include with the
amplifiable gene another marker, such as a drug resistance marker,
e.g. neomycin resistance (G418 in mammalian cells), hygromycin in
resistance etc., or an auxotrophy marker (HIS3, TRP1, LEU2, URA3,
ADE2, LYS2, etc.) for use in yeast cells.
[0318] Depending upon the nature of the modification and associated
targeting construct, various techniques may be employed for
identifying targeted integration. Conveniently, the DNA may be
digested with one or more restriction enzymes and the fragments
probed with an appropriate DNA fragment which will identify the
properly sized restriction fragment associated with
integration.
[0319] One may use different promoter sequences, enhancer
sequences, or other sequence which will allow for enhanced levels
of expression in the expression host. Thus, one may combine an
enhancer from one source, a promoter region from another source, a
5'-noncoding region upstream from the initiation methionine from
the same or different source as the other sequences and the like.
One may provide for an intron in the non-coding region with
appropriate splice sites or for an alternative 3'-untranslated
sequence or polyadenylation site. Depending upon the particular
purpose of the modification, any of these sequences may be
introduced, as desired.
[0320] Where selection is intended, the sequence to be integrated
will have with it a marker gene, which allows for selection. The
marker gene may conveniently be downstream from the target gene and
may include resistance to a cytotoxic agent, e.g. antibiotics,
heavy metals, or the like, resistance or susceptibility to HAT,
gancyclovir, etc., complementation to an auxotrophic host,
particularly by using an auxotrophic yeast as the host for the
subject manipulations, or the like. The marker gene may also be on
a separate DNA molecule, particularly with primary mammalian cells.
Alternatively, one may screen the various transformants, due to the
high efficiency of recombination in yeast, by using hybridization
analysis, PCR, sequencing, or the like.
[0321] For homologous recombination, constructs can be prepared
where the amplifiable gene will be flanked, normally on both sides
with DNA homologous with the DNA of the target region. Depending
upon the nature of the integrating DNA and the purpose of the
integration, the homologous DNA will generally be within 100 kb,
usually 50 kb, preferably about 25 kb, of the transcribed region of
the target gene, more preferably within 2 kb of the target gene.
Where modeling of the gene is intended, homology will usually be
present proximal to the site of the mutation. The homologous DNA
may include the 5'-upstream region outside of the transcriptional
regulatory region or comprising any enhancer sequences,
transcriptional initiation sequences, adjacent sequences, or the
like. The homologous region may include a portion of the coding
region, where the coding region may be comprised only of an open
reading frame or combination of exons and introns. The homologous
region may comprise all or a portion of an intron, where all or a
portion of one or more exons may also be present. Alternatively,
the homologous region may comprise the 3'-region, so as to comprise
all or a portion of the transcriptional termination region, or the
region 3' of this region. The homologous regions may extend over
all or a portion of the target gene or be outside the target gene
comprising all or a portion of the transcriptional regulatory
regions and/or the structural gene.
[0322] The integrating constructs may be prepared in accordance
with conventional ways, where sequences may be synthesized,
isolated from natural sources, manipulated, cloned, ligated,
subjected to in vitro mutagenesis, primer repair, or the like. At
various stages, the joined sequences may be cloned and analyzed by
restriction analysis, sequencing, or the like. Usually during the
preparation of a construct where various fragments are joined, the
fragments, intermediate constructs and constructs will be carried
on a cloning vector comprising a replication system functional in a
prokaryotic host, e.g., E. coli and a marker for selection, e.g.,
biocide resistance, complementation to an auxotrophic host, etc.
Other functional sequences may also be present, such as
polylinkers, for ease of introduction and excision of the construct
or portions thereof, or the like. A large number of cloning vectors
are available such as pBR322, the pUC series, etc. These constructs
may then be used for integration into the primary mammalian
host.
[0323] In the case of the primary mammalian host, a replicating
vector may be used. Usually, such vector will have a viral
replication system, such as SV40, bovine papilloma virus,
adenovirus, or the like. The linear DNA sequence vector may also
have a selectable marker for identifying transfected cells.
Selectable markers include the neo gene, allowing for selection
with G418, the herpes tk gene for selection with HAT medium, the
gpt gene with mycophenolic acid, complementation of an auxotrophic
host, etc.
[0324] The vector may or may not be capable of stable maintenance
in the host. Where the vector is capable of stable maintenance, the
cells will be screened for homologous integration of the vector
into the genome of the host, where various techniques for curing
the cells may be employed. Where the vector is not capable of
stable maintenance, for example, where a temperature sensitive
replication system is employed, one may change the temperature from
the permissive temperature to the non-permissive temperature, so
that the cells may be cured of the vector. In this case, only those
cells having integration of the construct comprising the
amplifiable gene and, when present, the selectable marker, will be
able to survive selection.
[0325] Where a selectable marker is present, one may select for the
presence of the targeting construct by means of the selectable
marker. Where the selectable marker is not present, one may select
for the presence of the construct by the amplifiable gene. For the
neo gene or the herpes tk gene, one could employ a medium for
growth of the transformants of about 0.1-1 mg/ml of G418 or may use
HAT medium, respectively. Where DHFR is the amplifiable gene, the
selective medium may include from about 0.01-0.5 .mu.M of
methotrexate or be deficient in glycine-hypoxanthine-thymidine and
have dialysed serum (GHT media).
[0326] The DNA can be introduced into the expression host by a
variety of techniques that include calcium phosphate/DNA
co-precipitates, microinjection of DNA into the nucleus,
electroporation, yeast protoplast fusion with intact cells,
transfection, polycations, e.g., polybrene, polyornithine, etc., or
the like. The DNA may be single or double stranded DNA, linear or
circular. The various techniques for transforming mammalian cells
are well known (see Keown et al., Methods Enzymol. (1989); Keown et
al., Methods Enzymol. 185:527-537 (1990); Mansour et al., Nature
336:348-352, (1988); all of which are herein incorporated by
reference in their entirety).
[0327] (d) Insect Constructs and Transformed Insect Cells
[0328] The present invention also relates to an insect recombinant
vectors comprising exogenous genetic material. The present
invention also relates to an insect cell comprising an insect
recombinant vector. The present invention also relates to methods
for obtaining a recombinant insect host cell, comprising
introducing into an insect cell exogenous genetic material.
[0329] The insect recombinant vector may be any vector which can be
conveniently subjected to recombinant DNA procedures and can bring
about the expression of the nucleic acid sequence. The choice of a
vector will typically depend on the compatibility of the vector
with the insect host cell into which the vector is to be
introduced. The vector may be a linear or a closed circular
plasmid. The vector system may be a single vector or plasmid or two
or more vectors or plasmids which together contain the total DNA to
be introduced into the genome of the insect host. In addition, the
insect vector may be an expression vector. Nucleic acid molecules
can be suitably inserted into a replication vector for expression
in the insect cell under a suitable promoter for insect cells. Many
vectors are available for this purpose and selection of the
appropriate vector will depend mainly on the size of the nucleic
acid molecule to be inserted into the vector and the particular
host cell to be transformed with the vector. Each vector contains
various components depending on its function (amplification of DNA
or expression of DNA) and the particular host cell with which it is
compatible. The vector components for insect cell transformation
generally include, but are not limited to, one or more of the
following: a signal sequence, origin of replication, one or more
marker genes and an inducible promoter.
[0330] The insect vector may be an autonomously replicating vector,
i.e., a vector which exists as an extrachromosomal entity, the
replication of which is independent of chromosomal replication,
e.g., a plasmid, an extrachromosomal element, a minichromosome, or
an artificial chromosome. The vector may contain any means for
assuring self-replication. Alternatively, the vector may be one
which, when introduced into the insect cell, is integrated into the
genome and replicated together with the chromosome(s) into which it
has been integrated. For integration, the vector may rely on the
nucleic acid sequence of the vector for stable integration of the
vector into the genome by homologous or nonhomologous
recombination. Alternatively, the vector may contain additional
nucleic acid sequences for directing integration by homologous
recombination into the genome of the insect host. The additional
nucleic acid sequences enable the vector to be integrated into the
host cell genome at a precise location(s) in the chromosome(s). To
increase the likelihood of integration at a precise location, there
should be preferably two nucleic acid sequences which individually
contain a sufficient number of nucleic acids, preferably 400 bp to
1500 bp, more preferably 800 bp to 1000 bp, which are highly
homologous with the corresponding target sequence to enhance the
probability of homologous recombination. These nucleic acid
sequences may be any sequence that is homologous with a target
sequence in the genome of the insect host cell and, furthermore,
may be non-encoding or encoding sequences.
[0331] Baculovirus expression vectors (BEVs) have become important
tools for the expression of foreign genes, both for basic research
and for the production of proteins with direct clinical
applications in human and veterinary medicine (Doerfler, Curr. Top.
Microbiol. Immunol. 131:51-68 (1968); Luckow and Summers,
Bio/Technology 6:47-55 (1988a); Miller, Annual Review of Microbiol.
42:177-199 (1988); Summers, Curr. Comm. Molecular Biology, Cold
Spring Harbor Press, Cold Spring Harbor, N.Y. (1988); all of which
are herein incorporated by reference in their entirety). BEVs are
recombinant insect viruses in which the coding sequence for a
chosen foreign gene has been inserted behind a baculovirus promoter
in place of the viral gene, e.g., polyhedrin (Smith and Summers,
U.S. Pat. No. 4,745,051, the entirety of which is incorporated
herein by reference).
[0332] The use of baculovirus vectors relies upon the host cells
being derived from Lepidopteran insects such as Spodoptera
frugiperda or Trichoplusia ni. The preferred Spodoptera frugiperda
cell line is the cell line Sf9. The Spodoptera frugiperda Sf9 cell
line was obtained from American Type Culture Collection (Manassas,
Va.) and is assigned accession number ATCC CRL 1711 (Summers and
Smith, A Manual of Methods for Baculovirus Vectors and Insect Cell
Culture Procedures, Texas Ag. Exper. Station Bulletin No. 1555
(1988), the entirety of which is herein incorporated by reference).
Other insect cell systems, such as the silkworm B. mori may also be
used.
[0333] The proteins expressed by the BEVs are, therefore,
synthesized, modified and transported in host cells derived from
Lepidopteran insects. Most of the genes that have been inserted and
produced in the baculovirus expression vector system have been
derived from vertebrate species. Other baculovirus genes in
addition to the polyhedrin promoter may be employed to advantage in
a baculovirus expression system. These include immediate-early
(alpha), delayed-early (.beta.), late (.gamma.), or very late
(delta), according to the phase of the viral infection during which
they are expressed. The expression of these genes occurs
sequentially, probably as the result of a "cascade" mechanism of
transcriptional regulation. (Guarino and Summers, J. Virol.
57:563-571 (1986); Guarino and Summers, J. Virol. 61:2091-2099
(1987); Guarino and Summers, Virol. 162:444-451 (1988); all of
which are herein incorporated by reference in their entirety).
[0334] Insect recombinant vectors are useful as intermediates for
the infection or transformation of insect cell systems. For
example, an insect recombinant vector containing a nucleic acid
molecule encoding a baculovirus transcriptional promoter followed
downstream by an insect signal DNA sequence is capable of directing
the secretion of the desired biologically active protein from the
insect cell. The vector may utilize a baculovirus transcriptional
promoter region derived from any of the over 500 baculoviruses
generally infecting insects, such as for example the Orders
Lepidoptera, Diptera, Orthoptera, Coleoptera and Hymenoptera,
including for example but not limited to the viral DNAs of
Autographa californica MNPV, Bombyx mori NPV, Trichoplusia ni MNPV,
Rachiplusia ou MNPV or Galleria mellonella MNPV, wherein said
baculovirus transcriptional promoter is a baculovirus
immediate-early gene IEl or IEN promoter; an immediate-early gene
in combination with a baculovirus delayed-early gene promoter
region selected from the group consisting of 39K and a HindIII-k
fragment delayed-early gene; or a baculovirus late gene promoter.
The immediate-early or delayed-early promoters can be enhanced with
transcriptional enhancer elements. The insect signal DNA sequence
may code for a signal peptide of a Lepidopteran adipokinetic
hormone precursor or a signal peptide of the Manduca sexta
adipokinetic hormone precursor (Summers, U.S. Pat. No. 5,155,037;
the entirety of which is herein incorporated by reference). Other
insect signal DNA sequences include a signal peptide of the
Orthoptera Schistocerca gregaria locust adipokinetic hormone
precurser and the Drosophila melanogaster cuticle genes CP1, CP2,
CP3 or CP4 or for an insect signal peptide having substantially a
similar chemical composition and function (Summers, U.S. Pat. No.
5,155,037).
[0335] Insect cells are distinctly different from animal cells.
Insects have a unique life cycle and have distinct cellular
properties such as the lack of intracellular plasminogen activators
in which are present in vertebrate cells. Another difference is the
high expression levels of protein products ranging from 1 to
greater than 500 mg/liter and the ease at which cDNA can be cloned
into cells (Frasier, In Vitro Cell. Dev. Biol. 25:225 (1989);
Summers and Smith, In: A Manual of Methods for Baculovirus Vectors
and Insect Cell Culture Procedures, Texas Ag. Exper. Station
Bulletin No. 1555 (1988), both of which are incorporated by
reference in their entirety).
[0336] Recombinant protein expression in insect cells is achieved
by viral infection or stable transformation. For viral infection,
the desired gene is cloned into baculovirus at the site of the
wild-type polyhedron gene (Webb and Summers, Technique 2:173
(1990); Bishop and Posse, Adv. Gene Technol. 1:55 (1990); both of
which are incorporated by reference in their entirety).
[0337] The polyhedron gene is a component of a protein coat in
occlusions which encapsulate virus particles. Deletion or insertion
in the polyhedron gene results the failure to form occlusion
bodies. Occlusion negative viruses are morphologically different
from occlusion positive viruses and enable one skilled in the art
to identify and purify recombinant viruses.
[0338] The vectors of present invention preferably contain one or
more selectable markers which permit easy selection of transformed
cells. A selectable marker is a gene the product of which provides,
for example biocide or viral resistance, resistance to heavy
metals, prototrophy to auxotrophs and the like. Selection may be
accomplished by co-transformation, e.g., as described in WO
91/17243, a nucleic acid sequence of the present invention may be
operably linked to a suitable promoter sequence. The promoter
sequence is a nucleic acid sequence which is recognized by the
insect host cell for expression of the nucleic acid sequence. The
promoter sequence contains transcription and translation control
sequences which mediate the expression of the protein or fragment
thereof. The promoter may be any nucleic acid sequence which shows
transcriptional activity in the insect host cell of choice and may
be obtained from genes encoding polypeptides either homologous or
heterologous to the host cell.
[0339] For example, a nucleic acid molecule encoding a protein or
fragment thereof may also be operably linked to a suitable leader
sequence. A leader sequence is a nontranslated region of a mRNA
which is important for translation by the fungal host. The leader
sequence is operably linked to the 5' terminus of the nucleic acid
sequence encoding the protein or fragment thereof. The leader
sequence may be native to the nucleic acid sequence encoding the
protein or fragment thereof or may be obtained from foreign
sources. Any leader sequence which is functional in the insect host
cell of choice may be used in the present invention.
[0340] A polyadenylation sequence may also be operably linked to
the 3' terminus of the nucleic acid sequence of the present
invention. The polyadenylation sequence is a sequence which when
transcribed is recognized by the insect host to add polyadenosine
residues to transcribed mRNA. The polyadenylation sequence may be
native to the nucleic acid sequence encoding the protein or
fragment thereof or may be obtained from foreign sources. Any
polyadenylation sequence which is functional in the fungal host of
choice may be used in the present invention.
[0341] To avoid the necessity of disrupting the cell to obtain the
protein or fragment thereof and to minimize the amount of possible
degradation of the expressed polypeptide within the cell, it is
preferred that expression of the polypeptide gene gives rise to a
product secreted outside the cell. To this end, the protein or
fragment thereof of the present invention may be linked to a signal
peptide linked to the amino terminus of the protein or fragment
thereof. A signal peptide is an amino acid sequence which permits
the secretion of the protein or fragment thereof from the insect
host into the culture medium. The signal peptide may be native to
the protein or fragment thereof of the invention or may be obtained
from foreign sources. The 5' end of the coding sequence of the
nucleic acid sequence of the present invention may inherently
contain a signal peptide coding region naturally linked in
translation reading frame with the segment of the coding region
which encodes the secreted protein or fragment thereof.
[0342] At present, a mode of achieving secretion of a foreign gene
product in insect cells is by way of the foreign gene's native
signal peptide. Because the foreign genes are usually from
non-insect organisms, their signal sequences may be poorly
recognized by insect cells and hence, levels of expression may be
suboptimal. However, the efficiency of expression of foreign gene
products seems to depend primarily on the characteristics of the
foreign protein. On average, nuclear localized or non-structural
proteins are most highly expressed, secreted proteins are
intermediate and integral membrane proteins are the least
expressed. One factor generally affecting the efficiency of the
production of foreign gene products in a heterologous host system
is the presence of native signal sequences (also termed
presequences, targeting signals, or leader sequences) associated
with the foreign gene. The signal sequence is generally coded by a
DNA sequence immediately following (5' to 3') the translation start
site of the desired foreign gene.
[0343] The expression dependence on the type of signal sequence
associated with a gene product can be represented by the following
example: If a foreign gene is inserted at a site downstream from
the translational start site of the baculovirus polyhedrin gene so
as to produce a fusion protein (containing the N-terminus of the
polyhedrin structural gene), the fused gene is highly expressed.
But less expression is achieved when a foreign gene is inserted in
a baculovirus expression vector immediately following the
transcriptional start site and totally replacing the polyhedrin
structural gene.
[0344] Insertions into the region -50 to -1 significantly alter
(reduce) steady state transcription which, in turn, reduces
translation of the foreign gene product. Use of the pVL941 vector
optimizes transcription of foreign genes to the level of the
polyhedrin gene transcription. Even though the transcription of a
foreign gene may be optimal, optimal translation may vary because
of several factors involving processing: signal peptide
recognition, mRNA and ribosome binding, glycosylation, disulfide
bond formation, sugar processing, oligomerization, for example.
[0345] The properties of the insect signal peptide are expected to
be more optimal for the efficiency of the translation process in
insect cells than those from vertebrate proteins. This phenomenon
can generally be explained by the fact that proteins secreted from
cells are synthesized as precursor molecules containing hydrophobic
N-terminal signal peptides. The signal peptides direct transport of
the select protein to its target membrane and are then cleaved by a
peptidase on the membrane, such as the endoplasmic reticulum, when
the protein passes through it.
[0346] Another exemplary insect signal sequence is the sequence
encoding for Drosophila cuticle proteins such as CP1, CP2, CP3 or
CP4 (Summers, U.S. Pat. No. 5,278,050; the entirety of which is
herein incorporated by reference). Most of a 9 kb region of the
Drosophila genome containing genes for the cuticle proteins has
been sequenced. Four of the five cuticle genes contains a signal
peptide coding sequence interrupted by a short intervening sequence
(about 60 base pairs) at a conserved site. Conserved sequences
occur in the 5' mRNA untranslated region, in the adjacent 35 base
pairs of upstream flanking sequence and at -200 base pairs from the
mRNA start position in each of the cuticle genes.
[0347] Standard methods of insect cell culture, cotransfection and
preparation of plasmids are set forth in Summers and Smith (Summers
and Smith, A Manual of Methods for Baculovirus Vectors and Insect
Cell Culture Procedures, Texas Agricultural Experiment Station
Bulletin No. 1555, Texas A&M University (1987)). Procedures for
the cultivation of viruses and cells are described in Volkman and
Summers, J. Virol 19:820-832 (1975) and Volkman et al., J. Virol
19:820-832 (1976); both of which are herein incorporated by
reference in their entirety.
[0348] (e) Bacterial Constructs and Transformed Bacterial Cells
[0349] The present invention also relates to a bacterial
recombinant vector comprising exogenous genetic material. The
present invention also relates to a bacteria cell comprising a
bacterial recombinant vector. The present invention also relates to
methods for obtaining a recombinant bacteria host cell, comprising
introducing into a bacterial host cell exogenous genetic
material.
[0350] The bacterial recombinant vector may be any vector which can
be conveniently subjected to recombinant DNA procedures. The choice
of a vector will typically depend on the compatibility of the
vector with the bacterial host cell into which the vector is to be
introduced. The vector may be a linear or a closed circular
plasmid. The vector system may be a single vector or plasmid or two
or more vectors or plasmids which together contain the total DNA to
be introduced into the genome of the bacterial host. In addition,
the bacterial vector may be an expression vector. Nucleic acid
molecules encoding protein homologues or fragments thereof can, for
example, be suitably inserted into a replicable vector for
expression in the bacterium under the control of a suitable
promoter for bacteria. Many vectors are available for this purpose
and selection of the appropriate vector will depend mainly on the
size of the nucleic acid to be inserted into the vector and the
particular host cell to be transformed with the vector. Each vector
contains various components depending on its function
(amplification of DNA or expression of DNA) and the particular host
cell with which it is compatible. The vector components for
bacterial transformation generally include, but are not limited to,
one or more of the following: a signal sequence, an origin of
replication, one or more marker genes and an inducible
promoter.
[0351] In general, plasmid vectors containing replicon and control
sequences that are derived from species compatible with the host
cell are used in connection with bacterial hosts. The vector
ordinarily carries a replication site, as well as marking sequences
that are capable of providing phenotypic selection in transformed
cells. For example, E. coli is typically transformed using pBR322,
a plasmid derived from an E. coli species (see, e.g., Bolivar et
al., Gene 2:95 (1977); the entirety of which is herein incorporated
by reference). pBR322 contains genes for ampicillin and
tetracycline resistance and thus provides easy means for
identifying transformed cells. The pBR322 plasmid, or other
microbial plasmid or phage, also generally contains, or is modified
to contain, promoters that can be used by the microbial organism
for expression of the selectable marker genes.
[0352] Nucleic acid molecules encoding protein or fragments thereof
may be expressed not only directly, but also as a fusion with
another polypeptide, preferably a signal sequence or other
polypeptide having a specific cleavage site at the N-terminus of
the mature polypeptide. In general, the signal sequence may be a
component of the vector, or it may be a part of the polypeptide DNA
that is inserted into the vector. The heterologous signal sequence
selected should be one that is recognized and processed (i.e.,
cleaved by a signal peptidase) by the host cell. For bacterial host
cells that do not recognize and process the native polypeptide
signal sequence, the signal sequence is substituted by a bacterial
signal sequence selected, for example, from the group consisting of
the alkaline phosphatase, penicillinase, 1 pp, or heat-stable
enterotoxin II leaders.
[0353] Both expression and cloning vectors contain a nucleic acid
sequence that enables the vector to replicate in one or more
selected host cells. Generally, in cloning vectors this sequence is
one that enables the vector to replicate independently of the host
chromosomal DNA and includes origins of replication or autonomously
replicating sequences. Such sequences are well known for a variety
of bacteria. The origin of replication from the plasmid pBR322 is
suitable for most Gram-negative bacteria.
[0354] Expression and cloning vectors also generally contain a
selection gene, also termed a selectable marker. This gene encodes
a protein necessary for the survival or growth of transformed host
cells grown in a selective culture medium. Host cells not
transformed with the vector containing the selection gene will not
survive in the culture medium. Typical selection genes encode
proteins that (a) confer resistance to antibiotics or other toxins,
e.g., ampicillin, neomycin, methotrexate, or tetracycline, (b)
complement auxotrophic deficiencies, or (c) supply critical
nutrients not available from complex media, e.g., the gene encoding
D-alanine racemase for Bacilli. One example of a selection scheme
utilizes a drug to arrest growth of a host cell. Those cells that
are successfully transformed with a heterologous protein homologue
or fragment thereof produce a protein conferring drug resistance
and thus survive the selection regimen.
[0355] The expression vector for producing a protein or fragment
thereof can also contains an inducible promoter that is recognized
by the host bacterial organism and is operably linked to the
nucleic acid encoding, for example, the nucleic acid molecule
encoding the protein homologue or fragment thereof of interest.
Inducible promoters suitable for use with bacterial hosts include
the .beta.-lactamase and lactose promoter systems (Chang et al.,
Nature 275:615 (1978); Goeddel et al., Nature 281:544 (1979); both
of which are herein incorporated by reference in their entirety),
the arabinose promoter system (Guzman et al., J. Bacteriol.
174:7716-7728 (1992); the entirety of which is herein incorporated
by reference), alkaline phosphatase, a tryptophan (trp) promoter
system (Goeddel, Nucleic Acids Res. 8:4057 (1980); EP 36,776; both
of which are herein incorporated by reference in their entirety)
and hybrid promoters such as the tac promoter (deBoer et al., Proc.
Natl. Acad. Sci. (USA) 80:21-25 (1983); the entirety of which is
herein incorporated by reference). However, other known bacterial
inducible promoters are suitable (Siebenlist et al., Cell 20:269
(1980); the entirety of which is herein incorporated by
reference).
[0356] Promoters for use in bacterial systems also generally
contain a Shine-Dalgarno (S.D.) sequence operably linked to the DNA
encoding the polypeptide of interest. The promoter can be removed
from the bacterial source DNA by restriction enzyme digestion and
inserted into the vector containing the desired DNA.
[0357] Construction of suitable vectors containing one or more of
the above-listed components employs standard ligation techniques.
Isolated plasmids or DNA fragments are cleaved, tailored and
re-ligated in the form desired to generate the plasmids required.
Examples of available bacterial expression vectors include, but are
not limited to, the multifunctional E. coli cloning and expression
vectors such as Bluescript.TM. (Stratagene, La Jolla, Calif.), in
which, for example, encoding an A. nidulans protein homologue or
fragment thereof homologue, may be ligated into the vector in frame
with sequences for the amino-terminal Met and the subsequent 7
residues of .beta.-galactosidase so that a hybrid protein is
produced; pIN vectors (Van Heeke and Schuster, J. Biol. Chem.
264:5503-5509 (1989), the entirety of which is herein incorporated
by reference); and the like. pGEX vectors (Promega, Madison Wis.
U.S.A.) may also be used to express foreign polypeptides as fusion
proteins with glutathione S-transferase (GST). In general, such
fusion proteins are soluble and can easily be purified from lysed
cells by adsorption to glutathione-agarose beads followed by
elution in the presence of free glutathione. Proteins made in such
systems are designed to include heparin, thrombin or factor XA
protease cleavage sites so that the cloned polypeptide of interest
can be released from the GST moiety at will.
[0358] Suitable host bacteria for a bacterial vector include
archaebacteria and eubacteria, especially eubacteria and most
preferably Enterobacteriaceae. Examples of useful bacteria include
Escherichia, Enterobacter, Azotobacter, Erwinia, Bacillus,
Pseudomonas, Klebsiella, Proteus, Salmonella, Serratia, Shigella,
Rhizobia, Vitreoscilla and Paracoccus. Suitable E. coli hosts
include E. coli W3110 (American Type Culture Collection (ATCC)
27,325, Manassas, Va. U.S.A.), E. coli 294 (ATCC 31,446), E. coli B
and E. coli X1776 (ATCC 31,537). These examples are illustrative
rather than limiting. Mutant cells of any of the above-mentioned
bacteria may also be employed. It is, of course, necessary to
select the appropriate bacteria taking into consideration
replicability of the replicon in the cells of a bacterium. For
example, E. coli, Serratia, or Salmonella species can be suitably
used as the host when well known plasmids such as pBR322, pBR325,
pACYC177, or pKN410 are used to supply the replicon. E. coli strain
W3110 is a preferred host or parent host because it is a common
host strain for recombinant DNA product fermentations. Preferably,
the host cell should secrete minimal amounts of proteolytic
enzymes.
[0359] Host cells are transfected and preferably transformed with
the above-described vectors and cultured in conventional nutrient
media modified as appropriate for inducing promoters, selecting
transformants, or amplifying the genes encoding the desired
sequences.
[0360] Numerous methods of transfection are known to the ordinarily
skilled artisan, for example, calcium phosphate and
electroporation. Depending on the host cell used, transformation is
done using standard techniques appropriate to such cells. The
calcium treatment employing calcium chloride, as described in
section 1.82 of Sambrook et al., Molecular Cloning: A Laboratory
Manual, New York: Cold Spring Harbor Laboratory Press, (1989), is
generally used for bacterial cells that contain substantial
cell-wall barriers. Another method for transformation employs
polyethylene glycol/DMSO, as described in Chung and Miller (Chung
and Miller, Nucleic Acids Res. 16:3580 (1988); the entirety of
which is herein incorporated by reference). Yet another method is
the use of the technique termed electroporation.
[0361] Bacterial cells used to produce the polypeptide of interest
for purposes of this invention are cultured in suitable media in
which the promoters for the nucleic acid encoding the heterologous
polypeptide can be artificially induced as described generally,
e.g., in Sambrook et al., Molecular Cloning: A Laboratory Manual,
New York: Cold Spring Harbor Laboratory Press, (1989). Examples of
suitable media are given in U.S. Pat. Nos. 5,304,472 and 5,342,763;
both of which are incorporated by reference in their entirety.
[0362] In addition to the above discussed procedures, practitioners
are familiar with the standard resource materials which describe
specific conditions and procedures for the construction,
manipulation and isolation of macromolecules (e.g., DNA molecules,
plasmids, etc.), generation of recombinant organisms and the
screening and isolating of clones, (see for example, Sambrook et
al., Molecular Cloning: A Laboratory Manual, Cold Spring Harbor
Press (1989); Mailga et al., Methods in Plant Molecular Biology,
Cold Spring Harbor Press (1995), the entirety of which is herein
incorporated by reference; Birren et al., Genome Analysis:
Analyzing DNA, 1, Cold Spring Harbor, N.Y., the entirety of which
is herein incorporated by reference).
[0363] (f) Computer Readable Media
[0364] The nucleotide sequence provided in SEQ ID NO: 1 through SEQ
ID NO: 3204 or fragment thereof, or complement thereof, or a
nucleotide sequence at least 90% identical, preferably 95%,
identical even more preferably 99% or 100% identical to the
sequence provided in SEQ ID NO: 1 through SEQ ID NO: 3204 or
fragment thereof, or complement thereof, can be "provided" in a
variety of mediums to facilitate use. Such a medium can also
provide a subset thereof in a form that allows a skilled artisan to
examine the sequences.
[0365] A preferred subset of nucleotide sequences are those nucleic
acid sequences that encode a maize or soybean methionine
adenosyltransferase enzyme or complement thereof or fragment of
either, a nucleic acid molecule that encodes a maize or soybean
S-adenosylmethionine decarboxylase enzyme or complement thereof or
fragment of either, a nucleic acid molecule that encodes a maize or
soybean aspartate kinase enzyme or complement thereof or fragment
of either, a nucleic acid molecule that encodes a maize or soybean
aspartate-semialdehyde dehydrogenase enzyme or complement thereof
or fragment of either, a nucleic acid molecule that encodes a maize
or soybean O-succinylhomoserine (thiol)-lyase enzyme or complement
thereof or fragment of either, a nucleic acid molecule that encodes
a maize or soybean cystathionine .beta.-lyase enzyme or complement
thereof or fragment of either, a nucleic acid molecule that encodes
a maize or soybean
5-methyltetrahydropteroyltriglutamate-homocysteine-S-methyltransf-
erase enzyme or complement thereof or fragment of either, a nucleic
acid molecule that encodes a maize or a soybean
adenosyihomocysteinase enzyme or complement thereof or fragment of
either, a nucleic acid molecule that encodes a maize or a soybean
cystathionine .beta.-synthase enzyme or complement thereof or
fragment of either, a nucleic acid molecule that encodes a maize or
a soybean cytsathionine .gamma.-lyase enzyme or complement thereof
or fragment of either and a nucleic acid molecule that encodes a
maize or a soybean O-acetylhomoserine (thiol)-lyase enzyme or
complement thereof or fragment of either.
[0366] A further preferred subset of nucleic acid sequences is
where the subset of sequences is two proteins or fragments thereof,
more preferably three proteins or fragments thereof and even more
preferable four proteins or fragments thereof, these nucleic acid
sequences are selected from the group that comprises a maize or
soybean methionine adenosyltransferase enzyme or complement thereof
or fragment of either, a nucleic acid molecule that encodes a maize
or soybean S-adenosylmethionine decarboxylase enzyme or complement
thereof or fragment of either, a nucleic acid molecule that encodes
a maize or soybean aspartate kinase enzyme or complement thereof or
fragment of either, a nucleic acid molecule that encodes a maize or
soybean aspartate-semialdehyde dehydrogenase enzyme or complement
thereof or fragment of either, a nucleic acid molecule that encodes
a maize or soybean O-succinylhomoserine (thiol)-lyase enzyme or
complement thereof or fragment of either, a nucleic acid molecule
that encodes a maize or soybean cystathionine .beta.-lyase enzyme
or complement thereof or fragment of either, a nucleic acid
molecule that encodes a maize or soybean
5-methyltetrahydropteroyltriglutamate-homocysteine-S-methyltransf-
erase enzyme or complement thereof or fragment of either, a nucleic
acid molecule that encodes a maize or a soybean
adenosylhomocysteinase enzyme or complement thereof or fragment of
either, a nucleic acid molecule that encodes a maize or a soybean
cystathionine .beta.-synthase enzyme or complement thereof or
fragment of either, a nucleic acid molecule that encodes a maize or
a soybean cytsathionine .gamma.-lyase enzyme or complement thereof
or fragment of either and a nucleic acid molecule that encodes a
maize or a soybean O-acetylhomoserine (thiol)-lyase enzyme or
complement thereof or fragment of either.
[0367] In one application of this embodiment, a nucleotide sequence
of the present invention can be recorded on computer readable
media. As used herein, "computer readable media" refers to any
medium that can be read and accessed directly by a computer. Such
media include, but are not limited to: magnetic storage media, such
as floppy discs, hard disc, storage medium and magnetic tape:
optical storage media such as CD-ROM; electrical storage media such
as RAM and ROM; and hybrids of these categories such as
magnetic/optical storage media. A skilled artisan can readily
appreciate how any of the presently known computer readable mediums
can be used to create a manufacture comprising computer readable
medium having recorded thereon a nucleotide sequence of the present
invention.
[0368] As used herein, "recorded" refers to a process for storing
information on computer readable medium. A skilled artisan can
readily adopt any of the presently known methods for recording
information on computer readable medium to generate media
comprising the nucleotide sequence information of the present
invention. A variety of data storage structures are available to a
skilled artisan for creating a computer readable medium having
recorded thereon a nucleotide sequence of the present invention.
The choice of the data storage structure will generally be based on
the means chosen to access the stored information. In addition, a
variety of data processor programs and formats can be used to store
the nucleotide sequence information of the present invention on
computer readable medium. The sequence information can be
represented in a word processing text file, formatted in
commercially-available software such as WordPerfect and Microsoft
Word, or represented in the form of an ASCII file, stored in a
database application, such as DB2, Sybase, Oracle, or the like. A
skilled artisan can readily adapt any number of data processor
structuring formats (e.g. text file or database) in order to obtain
computer readable medium having recorded thereon the nucleotide
sequence information of the present invention.
[0369] By providing one or more of nucleotide sequences of the
present invention, a skilled artisan can routinely access the
sequence information for a variety of purposes. Computer software
is publicly available which allows a skilled artisan to access
sequence information provided in a computer readable medium. The
examples which follow demonstrate how software which implements the
BLAST (Altschul et al., J. Mol. Biol. 215:403-410 (1990), the
entirety of which is herein incorporated by reference) and BLAZE
(Brutlag et al., Comp. Chem. 17:203-207 (1993), the entirety of
which is herein incorporated by reference) search algorithms on a
Sybase system can be used to identify open reading frames (ORFs)
within the genome that contain homology to ORFs or proteins from
other organisms. Such ORFs are protein-encoding fragments within
the sequences of the present invention and are useful in producing
commercially important proteins such as enzymes used in amino acid
biosynthesis, metabolism, transcription, translation, RNA
processing, nucleic acid and a protein degradation, protein
modification and DNA replication, restriction, modification,
recombination and repair.
[0370] The present invention further provides systems, particularly
computer-based systems, which contain the sequence information
described herein. Such systems are designed to identify
commercially important fragments of the nucleic acid molecule of
the present invention. As used herein, "a computer-based system"
refers to the hardware means, software means and data storage means
used to analyze the nucleotide sequence information of the present
invention. The minimum hardware means of the computer-based systems
of the present invention comprises a central processing unit (CPU),
input means, output means and data storage means. A skilled artisan
can readily appreciate that any one of the currently available
computer-based system are suitable for use in the present
invention.
[0371] As indicated above, the computer-based systems of the
present invention comprise a data storage means having stored
therein a nucleotide sequence of the present invention and the
necessary hardware means and software means for supporting and
implementing a search means. As used herein, "data storage means"
refers to memory that can store nucleotide sequence information of
the present invention, or a memory access means which can access
manufactures having recorded thereon the nucleotide sequence
information of the present invention. As used herein, "search
means" refers to one or more programs which are implemented on the
computer-based system to compare a target sequence or target
structural motif with the sequence information stored within the
data storage means. Search means are used to identify fragments or
regions of the sequence of the present invention that match a
particular target sequence or target motif. A variety of known
algorithms are disclosed publicly and a variety of commercially
available software for conducting search means are available can be
used in the computer-based systems of the present invention.
Examples of such software include, but are not limited to,
MacPattern (EMBL), BLASTIN and BLASTIX (NCBIA). One of the
available algorithms or implementing software packages for
conducting homology searches can be adapted for use in the present
computer-based systems.
[0372] The most preferred sequence length of a target sequence is
from about 10 to 100 amino acids or from about 30 to 300 nucleotide
residues. However, it is well recognized that during searches for
commercially important fragments of the nucleic acid molecules of
the present invention, such as sequence fragments involved in gene
expression and protein processing, may be of shorter length.
[0373] As used herein, "a target structural motif," or "target
motif," refers to any rationally selected sequence or combination
of sequences in which the sequences the sequence(s) are chosen
based on a three-dimensional configuration which is formed upon the
folding of the target motif. There are a variety of target motifs
known in the art. Protein target motifs include, but are not
limited to, enzymatic active sites and signal sequences. Nucleic
acid target motifs include, but are not limited to, promoter
sequences, cis elements, hairpin structures and inducible
expression elements (protein binding sequences).
[0374] Thus, the present invention further provides an input means
for receiving a target sequence, a data storage means for storing
the target sequences of the present invention sequence identified
using a search means as described above and an output means for
outputting the identified homologous sequences. A variety of
structural formats for the input and output means can be used to
input and output information in the computer-based systems of the
present invention. A preferred format for an output means ranks
fragments of the sequence of the present invention by varying
degrees of homology to the target sequence or target motif. Such
presentation provides a skilled artisan with a ranking of sequences
which contain various amounts of the target sequence or target
motif and identifies the degree of homology contained in the
identified fragment.
[0375] A variety of comparing means can be used to compare a target
sequence or target motif with the data storage means to identify
sequence fragments sequence of the present invention. For example,
implementing software which implement the BLAST and BLAZE
algorithms (Altschul et al., J. Mol. Biol. 215:403-410 (1990)) can
be used to identify open frames within the nucleic acid molecules
of the present invention. A skilled artisan can readily recognize
that any one of the publicly available homology search programs can
be used as the search means for the computer-based systems of the
present invention.
[0376] Having now generally described the invention, the same will
be more readily understood through reference to the following
examples which are provided by way of illustration and are not
intended to be limiting of the present invention, unless
specified.
Example 1
[0377] The MONN01 cDNA library is a normalized library generated
from maize (DK604, Dekalb Genetics, Dekalb, Ill. U.S.A.) total leaf
tissue at the V6 plant development stage. Seeds are planted at a
depth of approximately 3 cm into 2-3 inch peat pots containing
Metro 200 growing medium. After 2-3 weeks growth they are
transplanted into 10 inch pots containing the same growing medium.
Plants are watered daily before transplantation and three times a
week after transplantation. Peters 15-16-17 fertilizer is applied
three times per week after transplanting at a strength of 150 ppm
N. Two to three times during the lifetime of the plant, from
transplanting to flowering, a total of 900 mg Fe is added to each
pot. Maize plants are grown in the greenhouse in 15 hr day/9 hr
night cycles. The daytime temperature is approximately 80.degree.
F. and the nighttime temperature is approximately 70.degree. F.
Supplemental lighting is provided by 1000 W sodium vapor lamps.
Tissue is collected when the maize plant is at the 6-leaf
development stage. The older, more juvenile leaves, which are in a
basal position, as well as the younger, more adult leaves, which
are more apical are cut at the base of the leaves. The leaves are
then pooled and immediately transferred to liquid nitrogen
containers in which the pooled leaves are crushed. The harvested
tissue is then stored at -80.degree. C. until RNA preparation.
[0378] The SATMON001 cDNA library is generated from maize (B73,
Illinois Foundation Seeds, Champaign, Ill. U.S.A.) immature tassels
at the V6 plant development stage. Seeds are planted at a depth of
approximately 3 cm into 2-3 inch peat pots containing Metro 200
growing medium. After 2-3 weeks growth they are transplanted into
10 inch pots containing the same growing medium. Plants are watered
daily before transplantation and three times a week after
transplantation. Peters 15-16-17 fertilizer is applied three times
per week after transplanting at a strength of 150 ppm N. Two to
three times during the lifetime of the plant, from transplanting to
flowering, a total of 900 mg Fe is added to each pot. Maize plants
are grown in a greenhouse in 15 hr day/9 hr night cycles. The
daytime temperature is approximately 80.degree. F. and the
nighttime temperature is approximately 70.degree. F. Supplemental
lighting is provided by 1000 W sodium vapor lamps. Tissue from the
maize plant is collected at the V6 stage. At that stage the tassel
is an immature tassel of about 2-3 cm in length. The tassels are
removed and frozen in liquid nitrogen. The harvested tissue is then
stored at -80.degree. C. until RNA preparation.
[0379] The SATMON003 library is generated from maize
(B73.times.Mo17, Illinois Foundation Seeds, Champaign, Ill. U.S.A.)
roots at the V6 developmental stage. Seeds are planted at a depth
of approximately 3 cm in coil into 2-3 inch peat pots containing
Metro 200 growing medium. After 2-3 weeks growth, the seedlings are
transplanted into 10 inch pots containing the Metro 200 growing
medium. Plants are watered daily before transplantation and
approximately 3 times a week after transplantation. Peters 15-16-17
fertilizer is applied approximately three times per week after
transplanting at a concentration of 150 ppm N. Two to three times
during the life time of the plant from transplanting to flowering a
total of approximately 900 mg Fe is added to each pot. Maize plants
are grown in the green house in approximately 15 hr day/9 hr night
cycles. The daytime temperature is approximately 80.degree. F. and
the nighttime temperature is approximately 70.degree. F.
Supplemental lighting is provided by 1000 W sodium vapor lamps.
Tissue is collected when the maize plant is at the 6 leaf
development stage. The root system is cut from maize plant and
washed with water to free it from the soil. The tissue is then
immediately frozen in liquid nitrogen. The harvested tissue is then
stored at -80.degree. C. until RNA preparation.
[0380] The SATMON004 cDNA library is generated from maize
(B73.times.Mo17, Illinois Foundation Seeds, Champaign, Ill. U.S.A.)
total leaf tissue at the V6 plant development stage. Seeds are
planted at a depth of approximately 3 cm into 2-3 inch peat pots
containing Metro 200 growing medium. After 2-3 weeks growth they
are transplanted into 10 inch pots containing the same growing
medium. Plants are watered daily before transplantation and three
times a week after transplantation. Peters 15-16-17 fertilizer is
applied three times per week after transplanting at a strength of
150 ppm N. Two to three times during the lifetime of the plant,
from transplanting to flowering, a total of 900 mg Fe is added to
each pot. Maize plants are grown in the greenhouse in 15 hr day/9
hr night cycles. The daytime temperature is approximately
80.degree. F. and the nighttime temperature is approximately
70.degree. F. Supplemental lighting is provided by 1000 W sodium
vapor lamps. Tissue is collected when the maize plant is at the
6-leaf development stage. The older, more juvenile leaves, which
are in a basal position, as well as the younger, more adult leaves,
which are more apical are cut at the base of the leaves. The leaves
are then pooled and immediately transferred to liquid nitrogen
containers in which the pooled leaves are crushed. The harvested
tissue is then stored at -80.degree. C. until RNA preparation.
[0381] The SATMON005 cDNA library is generated from maize
(B73.times.Mo17, Illinois Foundation Seeds, Champaign Ill., U.S.A.)
root tissue at the V6 development stage. Seeds are planted at a
depth of approximately 3 cm into 2-3 inch peat pots containing
Metro 200 growing medium. After 2-3 weeks growth they are
transplanted into 10 inch pots containing the same growing medium.
Plants are watered daily before transplantation and three times a
week after transplantation. Peters 15-16-17 fertilizer is applied
three times per week after transplanting at a strength of 150 ppm
N. Two to three times during the lifetime of the plant, from
transplanting to flowering, a total of 900 mg Fe is added to each
pot. Maize plants are grown in the green house in 15 hr day/9 hr
night cycles. The daytime temperature is approximately 80.degree.
F. and the nighttime temperature is approximately 70.degree. F.
Supplemental lighting is provided by 1000 W sodium vapor lamps.
Tissue is collected when the maize plant is at the 6-leaf
development stage. The root system is cut from the mature maize
plant and washed with water to free it from the soil. The tissue is
immediately frozen in liquid nitrogen and the harvested tissue is
then stored at -80.degree. C. until RNA preparation.
[0382] The SATMON006 cDNA library is generated from maize
(B73.times.Mo17, Illinois Foundation Seeds, Champaign Ill., U.S.A.)
total leaf tissue at the V6 plant development stage. Seeds are
planted at a depth of approximately 3 cm into 2-3 inch peat pots
containing Metro 200 growing medium. After 2-3 weeks growth they
are transplanted into 10 inch pots containing the same growing
medium. Plants are watered daily before transplantation and three
times a week after transplantation. Peters 15-16-17 fertilizer is
applied three times per week after transplanting at a strength of
150 ppm N. Two to three times during the lifetime of the plant,
from transplanting to flowering, a total of 900 mg Fe is added to
each pot. Maize plants are grown in the greenhouse in 15 hr day/9
hr night cycles. The daytime temperature is approximately
80.degree. F. and the nighttime temperature is approximately
70.degree. F. Supplemental lighting is provided by 1000 W sodium
vapor lamps. Tissue is collected when the maize plant is at the
6-leaf development stage. The older more juvenile leaves, which are
in a basal position, as well as the younger more adult leaves,
which are more apical are cut at the base of the leaves. The leaves
are then pooled and immediately transferred to liquid nitrogen
containers in which the pooled leaves are crushed. The harvested
tissue is then stored at -80.degree. C. until RNA preparation.
[0383] The SATMON007 cDNA library is generated from the primary
root tissue of 5 day old maize (DK604, Dekalb Genetics, Dekalb,
Ill. U.S.A.) seedlings. Seeds are planted on a moist filter paper
on a covered tray that is kept in the dark until germination (one
day). After germination, the trays, along with the moist paper, are
moved to a greenhouse where the maize plants are grown in the
greenhouse in 15 hr day/9 hr night cycles for approximately 5 days.
The daytime temperature is approximately 80.degree. F. and the
nighttime temperature is approximately 70.degree. F. Supplemental
lighting is provided by 1000 W sodium vapor lamps. The primary root
tissue is collected when the seedlings are 5 days old. At this
stage, the primary root (radicle) is pushed through the coleorhiza
which itself is pushed through the seed coat. The primary root,
which is about 2-3 cm long, is cut and immediately frozen in liquid
nitrogen and then stored at -80.degree. C. until RNA
preparation.
[0384] The SATMON008 cDNA library is generated from the primary
shoot (coleoptile 2-3 cm) of maize (DK604, Dekalb Genetics, Dekalb,
Ill. U.S.A.) seedlings which are approximately 5 days old. Seeds
are planted on a moist filter paper on a covered tray that is kept
in the dark until germination (one day). Then the trays containing
the seeds are moved to a greenhouse at 15 hr daytime/9 hr nighttime
cycles and grown until they are 5 days post germination. The
daytime temperature is approximately 80.degree. F. and the
nighttime temperature is approximately 70.degree. F. Tissue is
collected when the seedlings are 5 days old. At this stage, the
primary shoot (coleoptile) is pushed through the seed coat and is
about 2-3 cm long. The coleoptile is dissected away from the rest
of the seedling, immediately frozen in liquid nitrogen and then
stored at -80.degree. C. until RNA preparation.
[0385] The SATMON009 cDNA library is generated from maize (DK604,
Dekalb Genetics, Dekalb, Ill. U.S.A.) leaves at the 8 leaf stage
(V8 plant development stage). Seeds are planted at a depth of
approximately 3 cm into 2-3 inch peat pots containing Metro 200
growing medium. After 2-3 weeks growth they are transplanted into
10 inch pots containing the same growing medium. Plants are watered
daily before transplantation and three times a week after
transplantation. Peters 15-16-17 fertilizer is applied three times
per week after transplanting at a strength of 150 ppm N. Two to
three times during the lifetime of the plant, from transplanting to
flowering, a total of 900 mg Fe is added to each pot. Maize plants
are grown in the green house in 15 hr day/9 hr night cycles. The
daytime temperature is 80.degree. F. and the nighttime temperature
is 70.degree. F. Supplemental lighting is provided by 1000 W sodium
vapor lamps. Tissue is collected when the maize plant is at the
8-leaf development stage. The older more juvenile leaves, which are
in a basal position, as well as the younger more adult leaves,
which are more apical, are cut at the base of the leaves. The
leaves are then pooled and then immediately transferred to liquid
nitrogen containers in which the pooled leaves are crushed. The
harvested tissue is then stored at -80.degree. C. until RNA
preparation.
[0386] The SATMON010 cDNA library is generated from maize (DK604,
Dekalb Genetics, Dekalb, Ill. U.S.A.) root tissue at the V8 plant
development stage. Seeds are planted at a depth of approximately 3
cm into 2-3 inch peat pots containing Metro 200 growing medium.
After 2-3 weeks growth they are transplanted into 10 inch pots
containing the same growing medium. Plants are watered daily before
transplantation and three times a week after transplantation.
Peters 15-16-17 fertilizer is applied three times per week after
transplanting at a strength of 150 ppm N. Two to three times during
the lifetime of the plant, from transplanting to flowering, a total
of 900 mg Fe is added to each pot. Maize plants are grown in the
green house in 15 hr day/9 hr night cycles. The daytime temperature
is 80.degree. F. and the nighttime temperature is 70.degree. F.
Supplemental lighting is provided by 1000 W sodium vapor lamps.
Tissue is collected when the maize plant is at the V8 development
stage. The root system is cut from this mature maize plant and
washed with water to free it from the soil. The tissue is
immediately frozen in liquid nitrogen. The harvested tissue is then
stored at -80.degree. C. until RNA preparation.
[0387] The SATMON011 cDNA library is generated from undeveloped
maize (DK604, Dekalb Genetics, Dekalb, Ill. U.S.A.) leaf at the V6
plant development stage. Seeds are planted at a depth of
approximately 3 cm into 2-3 inch peat pots containing Metro 200
growing medium. After 2-3 weeks growth they are transplanted into
10 inch pots containing the same growing medium. Plants are watered
daily before transplantation and three times a week after
transplantation. Peters 15-16-17 fertilizer is applied three times
per week after transplanting at a strength of 150 ppm N. Two to
three times during the lifetime of the plant, from transplanting to
flowering, a total of 900 mg Fe is added to each pot. Maize plants
are grown in the green house in 15 hr day/9 hr night cycles. The
daytime temperature is approximately 80.degree. F. and the
nighttime temperature is approximately 70.degree. F. Supplemental
lighting is provided by 1000 W sodium vapor lamps. Tissue is
collected when the maize plant is at the 6-leaf development stage.
The second youngest leaf which is at the base of the apical leaf of
V6 stage maize plant is cut at the base and immediately transferred
to liquid nitrogen containers in which the leaf is crushed. The
harvested tissue is then stored at -80.degree. C. until RNA
preparation.
[0388] The SATMON012 cDNA library is generated from 2 day post
germination maize (DK604, Dekalb Genetics, Dekalb, Ill. U.S.A.)
seedlings. Seeds are planted on a moist filter paper on a covered
tray that is kept in the dark until germination (one day). Then the
trays containing the seeds are moved to the greenhouse and grown at
15 hr daytime/9 hr nighttime cycles until 2 days post germination.
The daytime temperature is approximately 80.degree. F. and the
nighttime temperature is approximately 70.degree. F. Tissue is
collected when the seedlings are 2 days old. At the two day stage,
the coleorhiza is pushed through the seed coat and the primary root
(the radicle) is pierced the coleorhiza but is barely visible.
Also, at this two day stage, the coleoptile is just emerging from
the seed coat. The 2 days post germination seedlings are then
immersed in liquid nitrogen and crushed. The harvested tissue is
stored at -80.degree. C. until preparation of total RNA.
[0389] The SATMON013 cDNA library is generated from apical maize
(DK604, Dekalb Genetics, Dekalb, Ill. U.S.A.) meristem founder at
the V4 plant development stage. Seeds are planted at a depth of
approximately 3 cm into 2-3 inch peat pots containing Metro 200
growing medium. After 2-3 weeks growth they are transplanted into
10 inch pots containing the same growing medium. Plants are watered
daily before transplantation and three times a week after
transplantation. Peters 15-16-17 fertilizer is applied three times
per week after transplanting at a strength of 150 ppm N. Two to
three times during the lifetime of the plant, from transplanting to
flowering, a total of 900 mg Fe is added to each pot. Maize plants
are grown in the greenhouse in 15 hr day/9 hr night cycles. The
daytime temperature is approximately 80.degree. F. and the
nighttime temperature is approximately 70.degree. F. Supplemental
lighting is provided by 1000 W sodium vapor lamps. Prior to tissue
collection, the plant is at the 4 leaf stage. The lead at the apex
of the V4 stage maize plant is referred to as the meristem founder.
This apical meristem founder is cut, immediately frozen in liquid
nitrogen and crushed. The harvested tissue is then stored at
-80.degree. C. until RNA preparation.
[0390] The SATMON014 cDNA library is generated from maize (DK604,
Dekalb Genetics, Dekalb, Ill. U.S.A.) endosperm fourteen days after
pollination. Seeds are planted at a depth of approximately 3 cm
into 2-3 inch peat pots containing Metro 200 growing medium. After
2-3 weeks growth they are transplanted into 10 inch pots containing
the same growing medium. Plants are watered daily before
transplantation and three times a week after transplantation.
Peters 15-16-17 fertilizer is applied three times per week after
transplanting at a strength of 150 ppm N. Two to three times during
the lifetime of the plant, from transplanting to flowering, a total
of 900 mg Fe is added to each pot. Maize plants are grown in the
greenhouse in 15 hr day/9 hr night cycles. The daytime temperature
is approximately 80.degree. F. and the nighttime temperature is
approximately 70.degree. F. Supplemental lighting is provided by
1000 W sodium vapor lamps. After the V10 stage, the maize plant ear
shoots are ready for fertilization. At this stage, the ear shoots
are enclosed in a paper bag before silk emergence to withhold the
pollen. The ear shoots are pollinated and 14 days after
pollination, the ears are pulled out and then the kernels are
plucked out of the ears. Each kernel is then dissected into the
embryo and the endosperm and the aleurone layer is removed. After
dissection, the endosperms are immediately frozen in liquid
nitrogen and then stored at -80.degree. C. until RNA
preparation.
[0391] The SATMON016 library is a maize (DK604, Dekalb Genetics,
Dekalb, Ill. U.S.A.) sheath library collected at the V8
developmental stage. Seeds are planted in a depth of approximately
3 cm in solid into 2-3 inch pots containing Metro growing medium.
After 2-3 weeks growth, they are transplanted into 10'' pots
containing the same. Plants are watered daily before
transplantation and approximately the times a week after
transplantation. Peters 15-16-17 fertilizer is applied
approximately three times per week after transplanting, at a
strength of 150 ppm N. Two to three times during the life time of
the plant from transplanting to flowering, a total of approximately
900 mg Fe is added to each pot. Maize plants are grown in the green
house in 15 hr day/9 hr night cycles. The daytime temperature is
approximately 80.degree. F. and the nighttime temperature is
approximately 70.degree. F. Supplemental lighting is provided by
1000 W sodium vapor lamps. When the maize plants are at the V8
stage the 5.sup.th and 6.sup.th leaves from the bottom exhibit
fully developed leaf blades. At the base of these leaves, the
ligule is differentiated and the leaf blade is joined to the
sheath. The sheath is dissected away from the base of the leaf then
the sheath is frozen in liquid nitrogen and crushed. The tissue is
then stored at -80.degree. C. until RNA preparation.
[0392] The SATMON017 cDNA library is generated from maize (DK604,
Dekalb Genetics, Dekalb, Ill. U.S.A.) embryo seventeen days after
pollination. Seeds are planted at a depth of approximately 3 cm
into 2-3 inch peat pots containing Metro 200 growing medium. After
2-3 weeks growth the seeds are transplanted into 10 inch pots
containing the same growing medium. Plants are watered daily before
transplantation and three times a week after transplantation.
Peters 15-16-17 fertilizer is applied three times per week after
transplanting at a strength of 150 ppm N. Two to three times during
the lifetime of the plant, from transplanting to flowering, a total
of 900 mg Fe is added to each pot. Maize plants are grown in the
green house in 15 hr day/9 hr night cycles. The daytime temperature
is approximately 80.degree. F. and the nighttime temperature is
approximately 70.degree. F. Supplemental lighting is provided by
1000 W sodium vapor lamps. After the V10 stage, the ear shoots of
maize plant, which are ready for fertilization, are enclosed in a
paper bag before silk emergence to withhold the pollen. The ear
shoots are fertilized and 21 days after pollination, the ears are
pulled out and the kernels are plucked out of the ears. Each kernel
is then dissected into the embryo and the endosperm and the
aleurone layer is removed. After dissection, the embryos are
immediately frozen in liquid nitrogen and then stored at
-80.degree. C. until RNA preparation.
[0393] The SATMON019 (Lib3054) cDNA library is generated from maize
(DK604, Dekalb Genetics, Dekalb, Ill. U.S.A.) culm (stem) at the V8
developmental stage. Seeds are planted at a depth of approximately
3 cm into 2-3 inch peat pots containing Metro 200 growing medium.
After 2-3 weeks growth they are transplanted into 10 inch pots
containing the same growing medium. Plants are watered daily before
transplantation and three times a week after transplantation.
Peters 15-16-17 fertilizer is applied three times per week after
transplanting at a strength of 150 ppm N. Two to three times during
the lifetime of the plant, from transplanting to flowering, a total
of 900 mg Fe is added to each pot. Maize plants are grown in the
green house in 15 hr day/9 hr night cycles. The daytime temperature
is approximately 80.degree. F. and the nighttime temperature is
approximately 70.degree. F. Supplemental lighting is provided by
1000 W sodium vapor lamps. When the maize plant is at the V8 stage,
the 5th and 6th leaves from the bottom have fully developed leaf
blades. The region between the nodes of the 5th and the sixth
leaves from the bottom is the region of the stem that is collected.
The leaves are pulled out and the sheath is also torn away from the
stem. This stem tissue is completely free of any leaf and sheath
tissue. The stem tissue is then frozen in liquid nitrogen and
stored at -80.degree. C. until RNA preparation.
[0394] The SATMON020 cDNA library is from a maize (DK604, Dekalb
Genetics, Dekalb, Ill. U.S.A.) Hill Type II-Initiated Callus. Petri
plates containing approximately 25 ml of Type II initiation media
are prepared. This medium contains N6 salts and vitamins, 3%
sucrose, 2.3 g/liter proline 0.1 g/liter enzymatic casein
hydrolysate, 2 mg/liter 2,4-dichloro phenoxy-acetic acid (2,4, D),
15.3 mg/liter AgNO.sub.3 and 0.8% bacto agar and is adjusted to pH
6.0 before autoclaving. At 9-11 days after pollination, an ear with
immature embryos measuring approximately 1-2 mm in length is
chosen. The husks and silks are removed and then the ear is broken
into halves and placed in an autoclaved solution of Clorox/TWEEN 20
sterilizing solution. Then the ear is rinsed with deionized water.
Then each embryo is extracted from the kernel. Intact embryos are
placed in contact with the medium, scutellar side up). Multiple
embryos are plated on each plate and the plates are incubated in
the dark at 25.degree. C. Type II calluses are friable, can be
subcultured with a spatula, frequently regenerate via somatic
embryogenesis and are relatively undifferentiated. As seen in the
microscope, the Tape II calluses show color ranging from
translucent to light yellow and heterogeneity on with respect to
embryoid structure as well as stage of embryoid development. Once
Type II callus are formed, the calluses is transferred to type II
callus maintenance medium without AgN0.sub.3. Every 7-10 days, the
callus is subcultured. About 4 weeks after embryo isolation the
callus is removed from the plates and then frozen in liquid
nitrogen. The harvested tissue is stored at -80.degree. C. until
RNA preparation.
[0395] The SATMON021 cDNA library is generated from the immature
maize (DK604, Dekalb Genetics, Dekalb Ill., U.S.A.) tassel at the
V8 plant development stage. Seeds are planted at a depth of
approximately 3 cm into 2-3 inch peat pots containing Metro 200
growing medium. After 2-3 weeks growth they are transplanted into
10 inch pots containing the same growing medium. Plants are watered
daily before transplantation and three times a week after
transplantation. Peters 15-16-17 fertilizer is applied three times
per week after transplanting at a strength of 150 ppm N. Two to
three times during the lifetime of the plant, from transplanting to
flowering, a total of 900 mg Fe is added to each pot. Maize plants
are grown in the green house in 15 hr day/9 hr night cycles. The
daytime temperature is approximately 80.degree. F. and the
nighttime temperature is approximately 70.degree. F. Supplemental
lighting is provided by 1000 W sodium vapor lamps. As the maize
plant enters the V8 stage, tassels which are 15-20 cm in length are
collected and frozen in liquid nitrogen. The harvested tissue is
stored at -80.degree. C. until RNA preparation.
[0396] The SATMON022 cDNA library is generated from maize (DK604,
Dekalb Genetics, Dekalb, Ill. U.S.A.) ear (growing silks) at the V8
plant development stage. Seeds are planted at a depth of
approximately 3 cm into 2-3 inch peat pots containing Metro 200
growing medium. After 2-3 weeks growth they are transplanted into
10 inch pots containing the same growing medium. Plants are watered
daily before transplantation and three times a week after
transplantation. Peters 15-16-17 fertilizer is applied three times
per week after transplanting at a strength of 150 ppm N. Two to
three times during the lifetime of the plant, from transplanting to
flowering, a total of 900 mg Fe is added to each pot. Zea mays
plants are grown in the greenhouse in 15 hr day/9 hr night cycles.
The daytime temperature is approximately 80.degree. F. and the
nighttime temperature is approximately 70.degree. F. Supplemental
lighting is provided by 1000 W sodium vapor lamps. Tissue is
collected when the plant is in the V8 stage. At this stage, some
immature ear shoots are visible. The immature ear shoots
(approximately 1 cm in length) are pulled out, frozen in liquid
nitrogen and then stored at -80.degree. C. until RNA
preparation.
[0397] The SATMON23 cDNA library is generated from maize (DK604,
Dekalb Genetics, Dekalb, Ill. U.S.A.) ear (growing silk) at the V8
development stage. Seeds are planted at a depth of approximately 3
cm into 2-3 inch peat pots containing Metro 200 growing medium.
After 2-3 weeks growth they are transplanted into 10 inch pots
containing the same growing medium. Plants are watered daily before
transplantation and three times a week after transplantation.
Peters 15-16-17 fertilizer is applied three times per week after
transplanting at a strength of 150 ppm N. Two to three times during
the lifetime of the plant, from transplanting to flowering, a total
of 900 mg Fe is added to each pot. Maize plants are grown in the
greenhouse in 15 hr day/9 hr night cycles. The daytime temperature
is approximately 80.degree. F. and the nighttime temperature is
approximately 70.degree. F. When the tissue is harvested at the V8
stage, the length of the ear that is harvested is about 10-15 cm
and the silks are just exposed (approximately 1 inch). The ear
along with the silks is frozen in liquid nitrogen and then the
tissue is stored at -80.degree. C. until RNA preparation.
[0398] The SATMON024 cDNA library is generated from the immature
maize (DK604, Dekalb Genetics, Dekalb, Ill. U.S.A.) tassel at the
V9 development stage. Seeds are planted at a depth of approximately
3 cm into 2-3 inch peat pots containing Metro 200 growing medium.
After 2-3 weeks growth they are transplanted into 10 inch pots
containing the same growing medium. Plants are watered daily before
transplantation and three times a week after transplantation.
Peters 15-16-17 fertilizer is applied three times per week after
transplanting at a strength of 150 ppm N. Two to three times during
the lifetime of the plant, from transplanting to flowering, a total
of 900 mg Fe is added to each pot. Maize plants are grown in the
green house in 15 hr day/9 hr night cycles. The daytime temperature
is approximately 80.degree. F. and the nighttime temperature is
approximately 70.degree. F. As a maize plant enters the V9 stage,
the tassel is rapidly developing and a 37 cm tassel along with the
glume, anthers and pollen is collected and frozen in liquid
nitrogen. The harvested tissue is stored at -80.degree. C. until
RNA preparation.
[0399] The SATMON025 cDNA library is from maize (DK604, Dekalb
Genetics, Dekalb, Ill. U.S.A.) Hill Type II-Regenerated Callus.
Type II callus is grown in initiation media as described for
SATMON020 and then the embryoids on the surface of the Type II
callus are allowed to mature and germinate. The 1-2 gm fresh weight
of the soft friable type callus containing numerous embryoids are
transferred to 100.times.15 mm petri plates containing 25 ml of
regeneration media. Regeneration media consists of Murashige and
Skoog (MS) basal salts, modified White's vitamins (0.2 g/liter
glycine and 0.5 g/liter myo-inositoland 0.8% bacto agar (6SMS0D)).
The plates are then placed in the dark after covering with
parafilm. After 1 week, the plates are moved to a lighted growth
chamber with 16 hr light and 8 hr dark photoperiod. Three weeks
after plating the Type II callus to 6SMS0D, the callus exhibit
shoot formation. The callus and the shoots are transferred to fresh
6SMS0D plates for another 2 weeks. The callus and the shoots are
then transferred to petri plates with reduced sucrose (3SMSOD).
Upon distinct formation of a root and shoot, the newly developed
green plants are then removed out with a spatula and frozen in
liquid nitrogen containers. The harvested tissue is then stored at
-80.degree. C. until RNA preparation.
[0400] The SATMON026 cDNA library is generated from maize (DK604,
Dekalb Genetics, Dekalb, Ill. U.S.A.) juvenile/adult shift leaves
at the V8 plant development stage. Seeds are planted at a depth of
approximately 3 cm into 2-3 inch peat pots containing Metro 200
growing medium. After 2-3 weeks growth they are transplanted into
10 inch pots containing the same growing medium. Plants are watered
daily before transplantation and three times a week after
transplantation. Peters 15-16-17 fertilizer is applied three times
per week after transplanting at a strength of 150 ppm N. Two to
three times during the lifetime of the plant, from transplanting to
flowering, a total of 900 mg Fe is added to each pot. Maize plants
are grown in the green house in 15 hr day/9 hr night cycles. The
daytime temperature is approximately 80.degree. F. and the
nighttime temperature is approximately 70.degree. F. Supplemental
lighting is provided by 1000 W sodium vapor lamps. Tissue is
collected when the maize plants are at the 8-leaf development
stage. Leaves are founded sequentially around the meristem over
weeks of time and the older, more juvenile leaves arise earlier and
in a more basal position than the younger, more adult leaves, which
are in a more apical position. In a V8 plant, some leaves which are
in the middle portion of the plant exhibit characteristics of both
juvenile as well as adult leaves. They exhibit a yellowing color
but also exhibit, in part, a green color. These leaves are termed
juvenile/adult shift leaves. The juvenile/adult shift leaves (the
4th, 5th leaves from the bottom) are cut at the base, pooled and
transferred to liquid nitrogen in which they are then crushed. The
harvested tissue is then stored at -80.degree. C. until RNA
preparation.
[0401] The SATMON027 cDNA library is generated from 6 day maize
(DK604, Dekalb Genetics, Dekalb, Ill. U.S.A.) leaves. Seeds are
planted at a depth of approximately 3 cm into 2-3 inch peat pots
containing Metro 200 growing medium. After 2-3 weeks growth they
are transplanted into 10 inch pots containing the Metro 200 growing
medium. Plants are watered daily before transplantation and three
times a week after transplantation. Peters 15-16-17 fertilizer is
applied three times per week after transplanting at a strength of
150 ppm N. Two to three times during the lifetime of the plant,
from transplanting to flowering, a total of 900 mg Fe is added to
each pot. Zea mays plants are grown in the greenhouse in 15 hr
day/9 hr night cycles. The daytime temperature is approximately
80.degree. F. and the nighttime temperature is approximately
70.degree. F. Supplemental lighting is provided by 1000 W sodium
vapor lamps. Prior to tissue collection, when the plant is at the
8-leaf stage, water is held back for six days. The older, more
juvenile leaves, which are in a basal position, as well as the
younger, more adult leaves, which are more apical, are all cut at
the base of the leaves. All the leaves exhibit significant wilting.
The leaves are then pooled and immediately transferred to liquid
nitrogen containers in which the pooled leaves are then crushed.
The harvested tissue is then stored at -80.degree. C. until RNA
preparation.
[0402] The SATMON028 cDNA library is generated from maize (DK604,
Dekalb Genetics, Dekalb, Ill. U.S.A.) roots at the V8 developmental
stage that are subject to six days water stress. Seeds are planted
at a depth of approximately 3 cm into 2-3 inch peat pots containing
Metro 200 growing medium. After 2-3 weeks growth they are
transplanted into 10 inch pots containing the Metro 200 growing
medium. Plants are watered daily before transplantation and three
times a week after transplantation. Peters 15-16-17 fertilizer is
applied three times per week after transplanting at a strength of
150 ppm N. Two to three times during the lifetime of the plant,
from transplanting to flowering, a total of 900 mg Fe is added to
each pot. Maize plants are grown in the greenhouse in 15 hr day/9
hr night cycles. The daytime temperature is approximately
80.degree. F. and the nighttime temperature is approximately
70.degree. F. Supplemental lighting is provided by 1000 W sodium
vapor lamps. Prior to tissue collection, when the plant is at the
8-leaf stage, water is held back for six days. The root system is
cut, shaken and washed to remove soil. Root tissue is then pooled
and immediately transferred to liquid nitrogen containers in which
the pooled leaves are then crushed. The harvested tissue is then
stored at -80.degree. C. until RNA preparation.
[0403] The SATMON029 cDNA library is generated from maize (DK604,
Dekalb Genetics, Dekalb, Ill. U.S.A.) seedlings at the etiolated
stage. Seeds are planted on a moist filter paper on a covered tray
that is kept in the dark for 4 days at approximately 70.degree. F.
Tissue is collected when the seedlings are 4 days old. By 4 days,
the primary root has penetrated the coleorhiza and is about 4-5 cm
and the secondary lateral roots have also made their appearance.
The coleoptile has also pushed through the seed coat and is about
4-5 cm long. The seedlings are frozen in liquid nitrogen and
crushed. The harvested tissue is then stored at -80.degree. C.
until RNA preparation.
[0404] The SATMON030 cDNA library is generated from maize (DK604,
Dekalb Genetics, Dekalb, Ill. U.S.A.) root tissue at the V4 plant
development stage. Seeds are planted at a depth of approximately 3
cm into 2-3 inch peat pots containing Metro 200 growing medium.
After 2-3 weeks growth, they are transplanted into 10 inch pots
containing the same. Plants are watered daily before
transplantation and approximately 3 times a week after
transplantation. Peters 15-16-17 fertilizer is applied
approximately three times per week after transplanting, at a
strength of 150 ppm N. Two to three times during the life time of
the plant, from transplanting to flowering, a total of
approximately 900 mg Fe is added to each pot. Maize plants are
grown in the green house in 15 hr day/9 hr night cycles. The
daytime temperature is approximately 80.degree. F. and the
nighttime temperature is approximately 70.degree. F. Supplemental
lighting is provided by 1000 sodium vapor lamps. Tissue is
collected when the maize plant is at the 4 leaf development stage.
The root system is cut from the mature maize plant and washed with
water to free it from the soil. The tissue is then immediately
frozen in liquid nitrogen. The harvested tissue is then stored at
-80.degree. C. until RNA preparation.
[0405] The SATMON031 cDNA library is generated from the maize
(DK604, Dekalb Genetics, Dekalb, Ill. U.S.A.) leaf tissue at the V4
plant development stage. Seeds are planted at a depth of
approximately 3 cm into 2-3 inch peat pots containing Metro 200
growing medium. After 2-3 weeks growth they are transplanted into
10 inch pots containing the same growing medium. Plants are watered
daily before transplantation and three times a week after
transplantation. Peters 15-16-17 fertilizer is applied three times
per week after transplanting at a strength of 150 ppm N. Two to
three times during the lifetime of the plant, from transplanting to
flowering, a total of 900 mg Fe is added to each pot. Maize plants
are grown in the green house in 15 hr day/9 hr night cycles. The
daytime temperature is 80.degree. F. and the nighttime temperature
is 70.degree. F. Supplemental lighting is provided by 1000 W sodium
vapor lamps. Tissue is collected when the maize plant is at the
4-leaf development stage. The third leaf from the bottom is cut at
the base and immediately frozen in liquid nitrogen and crushed. The
tissue is immediately frozen in liquid nitrogen. The harvested
tissue is then stored at -80.degree. C. until RNA preparation.
[0406] The SATMON033 cDNA library is generated from maize (DK604,
Dekalb Genetics, Dekalb, Ill. U.S.A.) embryo tissue 13 days after
pollination. Seeds are planted at a depth of approximately 3 cm
into 2-3 inch peat pots containing Metro 200 growing medium. After
2-3 weeks growth they are transplanted into 10 inch pots containing
the same growing medium. Plants are watered daily before
transplantation and three times a week after transplantation.
Peters 15-16-17 fertilizer is applied three times per week after
transplanting at a strength of 150 ppm N. Two to three times during
the lifetime of the plant, from transplanting to flowering, a total
of 900 mg Fe is added to each pot. Maize plants are grown in the
greenhouse in 15 hr day/9 hr night cycles. The daytime temperature
is approximately 80.degree. F. and the nighttime temperature is
approximately 70.degree. F. Supplemental lighting is provided by
1000 W sodium vapor lamps. After the V10 stage, the ear shoots of
the maize plant, which are ready for fertilization, are enclosed in
a paper bag before silk emergent to withhold the pollen. The ear
shoots are pollinated and 13 days after pollination, the ears are
pulled out and then the kernels are plucked cut of the ears. Each
kernel is then dissected into the embryo and the endosperm and the
aleurone layer is removed. After dissection, the embryos are
immediately frozen in liquid nitrogen and then stored at
-80.degree. C. until RNA preparation.
[0407] The SATMON034 cDNA library is generated from cold stressed
maize (DK604, Dekalb Genetics, Dekalb, Ill. U.S.A.) seedlings.
Seeds are planted on a moist filter paper on a covered tray that is
kept on at 10.degree. C. for 7 days. After 7 days, the temperature
is shifted to 15.degree. C. for one day until germination of the
seed. Tissue is collected once the seedlings are 1 day old. At this
point, the coleorhiza has just pushed out of the seed coat and the
primary root is just making its appearance. The coleoptile has not
yet pushed completely through the seed coat and is also just making
its appearance. These 1 day old cold stressed seedlings are frozen
in liquid nitrogen and crushed. The harvested tissue is then stored
at -80.degree. C. until RNA preparation.
[0408] The SATMON.about.001 (Lib36, Lib83, Lib84) cDNA library is
generated from maize leaves at the V8 plant development stage.
Seeds are planted at a depth of approximately 3 cm into 2-3 inch
peat pots containing Metro 200 growing medium. After 2-3 weeks
growth they are transplanted into 10 inch pots containing the same
growing medium. Plants are watered daily before transplantation and
three times a week after transplantation. Peters 15-16-17
fertilizer is applied three times per week after transplanting at a
strength of 150 ppm N. Two to three times during the lifetime of
the plant, from transplanting to flowering, a total of 900 mg Fe is
added to each pot. Maize plants are grown in a greenhouse in 15 hr
day/9 hr night cycles. The daytime temperature is approximately
80.degree. F. and the nighttime temperature is approximately
70.degree. F. Supplemental lighting is provided by 1000 W sodium
vapor lamps. Tissue from the maize plant is collected at the V8
stage. The older more juvenile leaves in a basal position was well
as the younger more adult leaves which are more apical are all cut
at the base, pooled and frozen in liquid nitrogen. The harvested
tissue is then stored at -80.degree. C. until RNA preparation.
[0409] The SATMONN01 cDNA library is generated from maize (B73,
Illinois Foundation Seeds, Champaign, Ill. U.S.A.) normalized
immature tassels at the V6 plant development stage normalized
tissue. Seeds are planted at a depth of approximately 3 cm into 2-3
inch peat pots containing Metro 200 growing medium. After 2-3 weeks
growth they are transplanted into 10 inch pots containing the same
growing medium. Plants are watered daily before transplantation and
three times a week after transplantation. Peters 15-16-17
fertilizer is applied three times per week after transplanting at a
strength of 150 ppm N. Two to three times during the lifetime of
the plant, from transplanting to flowering, a total of 900 mg Fe is
added to each pot. Maize plants are grown in a greenhouse in 15 hr
day/9 hr night cycles. The daytime temperature is approximately
80.degree. F. and the nighttime temperature is approximately
70.degree. F. Supplemental lighting is provided by 1000 W sodium
vapor lamps. Tissue from the maize plant is collected at the V6
stage. At that stage the tassel is an immature tassel of about 2-3
cm in length. The tassels are removed and frozen in liquid
nitrogen. The harvested tissue is then stored at -80.degree. C.
until RNA preparation. Single stranded and double stranded DNA
representing approximately 1.times.10.sup.6 colony forming units
are isolated using standard protocols. RNA, complementary to the
single stranded DNA, is synthesized using the double stranded DNA
as a template. Biotinylated dATP is incorporated into the RNA
during the synthesis reaction. The single stranded DNA is mixed
with the biotinylated RNA in a 1:10 molar ratio) and allowed to
hybridize. DNA-RNA hybrids are captured on Dynabeads M280
streptavidin (Dynabeads, Dynal Corporation, Lake Success, N.Y.
U.S.A.). The dynabeads with captured hybrids are collected with a
magnet. The non-hybridized single stranded molecules remaining
after hybrid capture are converted to double stranded form and
represent the primary normalized library.
[0410] The SATMONN04 cDNA library is generated from maize
(B73.times.Mo17, Illinois Foundation Seeds, Champaign, Ill. U.S.A.)
normalized total leaf tissue at the V6 plant development stage.
Seeds are planted at a depth of approximately 3 cm into 2-3 inch
peat pots containing Metro 200 growing medium. After 2-3 weeks
growth they are transplanted into 10 inch pots containing the same
growing medium. Plants are watered daily before transplantation and
three times a week after transplantation. Peters 15-16-17
fertilizer is applied three times per week after transplanting at a
strength of 150 ppm N. Two to three times during the lifetime of
the plant, from transplanting to flowering, a total of 900 mg Fe is
added to each pot. Maize plants are grown in the greenhouse in 15
hr day/9 hr night cycles. The daytime temperature is approximately
80.degree. F. and the nighttime temperature is approximately
70.degree. F. Supplemental lighting is provided by 1000 W sodium
vapor lamps. Tissue is collected when the maize plant is at the
6-leaf development stage. The older, more juvenile leaves, which
are in a basal position, as well as the younger, more adult leaves,
which are more apical are cut at the base of the leaves. The leaves
are then pooled and immediately transferred to liquid nitrogen
containers in which the pooled leaves are crushed. The harvested
tissue is then stored at -80.degree. C. until RNA preparation.
Single stranded and double stranded DNA representing approximately
1.times.10.sup.6 colony forming units are isolated using standard
protocols. RNA, complementary to the single stranded DNA, is
synthesized using the double stranded DNA as a template.
Biotinylated dATP is incorporated into the RNA during the synthesis
reaction. The single stranded DNA is mixed with the biotinylated
RNA in a 1:10 molar ratio) and allowed to hybridize. DNA-RNA
hybrids are captured on Dynabeads M280 streptavidin (Dynabeads,
Dynal Corporation, Lake Success, N.Y. U.S.A.). The dynabeads with
captured hybrids are collected with a magnet. The non-hybridized
single stranded molecules remaining after hybrid capture are
converted to double stranded form and represent the primary
normalized library.
[0411] The SATMONN05 cDNA library is generated from maize
(B73.times.Mo17, Illinois Foundation Seeds, Champaign Ill., U.S.A.)
normalized root tissue at the V6 development stage. Seeds are
planted at a depth of approximately 3 cm into 2-3 inch peat pots
containing Metro 200 growing medium. After 2-3 weeks growth they
are transplanted into 10 inch pots containing the same growing
medium. Plants are watered daily before transplantation and three
times a week after transplantation. Peters 15-16-17 fertilizer is
applied three times per week after transplanting at a strength of
150 ppm N. Two to three times during the lifetime of the plant,
from transplanting to flowering, a total of 900 mg Fe is added to
each pot. Maize plants are grown in the green house in 15 hr day/9
hr night cycles. The daytime temperature is approximately
80.degree. F. and the nighttime temperature is approximately
70.degree. F. Supplemental lighting is provided by 1000 W sodium
vapor lamps. Tissue is collected when the maize plant is at the
6-leaf development stage. The root system is cut from the mature
maize plant and washed with water to free it from the soil. The
tissue is immediately frozen in liquid nitrogen and the harvested
tissue is then stored at -80.degree. C. until RNA preparation. The
single stranded and double stranded DNA representing approximately
1.times.10.sup.6 colony forming units are isolated using standard
protocols. RNA, complementary to the single stranded DNA, is
synthesized using the double stranded DNA as a template.
Biotinylated dATP is incorporated into the RNA during the synthesis
reaction. The single stranded DNA is mixed with the biotinylated
RNA in a 1:10 molar ratio) and allowed to hybridize. DNA-RNA
hybrids are captured on Dynabeads M280 streptavidin (Dynabeads,
Dynal Corporation, Lake Success, N.Y. U.S.A.). The dynabeads with
captured hybrids are collected with a magnet. The non-hybridized
single stranded molecules remaining after hybrid capture are
converted to double stranded form and represent the primary
normalized library.
[0412] The SATMONN06 cDNA library is generated from maize
(B73.times.Mo17, Illinois Foundation Seeds, Champaign Ill., U.S.A.)
normalized total leaf tissue at the V6 plant development stage.
Seeds are planted at a depth of approximately 3 cm into 2-3 inch
peat pots containing Metro 200 growing medium. After 2-3 weeks
growth they are transplanted into 10 inch pots containing the same
growing medium. Plants are watered daily before transplantation and
three times a week after transplantation. Peters 15-16-17
fertilizer is applied three times per week after transplanting at a
strength of 150 ppm N. Two to three times during the lifetime of
the plant, from transplanting to flowering, a total of 900 mg Fe is
added to each pot. Maize plants are grown in the greenhouse in 15
hr day/9 hr night cycles. The daytime temperature is approximately
80.degree. F. and the nighttime temperature is approximately
70.degree. F. Supplemental lighting is provided by 1000 W sodium
vapor lamps. Tissue is collected when the maize plant is at the
6-leaf development stage. The older more juvenile leaves, which are
in a basal position, as well as the younger more adult leaves,
which are more apical are cut at the base of the leaves. The leaves
are then pooled and immediately transferred to liquid nitrogen
containers in which the pooled leaves are crushed. The harvested
tissue is then stored at -80.degree. C. until RNA preparation.
Single stranded and double stranded DNA representing approximately
1.times.10.sup.6 colony forming units are isolated using standard
protocols. RNA, complementary to the single stranded DNA, is
synthesized using the double stranded DNA as a template.
Biotinylated dATP is incorporated into the RNA during the synthesis
reaction. The single stranded DNA is mixed with the biotinylated
RNA in a 1:10 molar ratio) and allowed to hybridize. DNA-RNA
hybrids are captured on Dynabeads M280 streptavidin (Dynabeads,
Dynal Corporation, Lake Success, N.Y. U.S.A.). The dynabeads with
captured hybrids are collected with a magnet. The non-hybridized
single stranded molecules remaining after hybrid capture are
converted to double stranded form and represent the primary
normalized library.
[0413] The CMZ029 (SATMON036) cDNA library is generated from maize
(DK604, Dekalb Genetics, Dekalb, Ill. U.S.A.) endosperm 22 days
after pollination. Seeds are planted at a depth of approximately 3
cm into 2-3 inch peat pots containing Metro 200 growing medium.
After 2-3 weeks growth they are transplanted into 10 inch pots
containing the same growing medium. Plants are watered daily before
transplantation and three times a week after transplantation.
Peters 15-16-17 fertilizer is applied three times per week after
transplanting at a strength of 150 ppm N. Two to three times during
the lifetime of the plant, from transplanting to flowering, a total
of 900 mg Fe is added to each pot. Maize plants are grown in the
green house in 15 hr day/9 hr night cycles. The daytime temperature
is approximately 80.degree. F. and the nighttime temperature is
approximately 70.degree. F. Supplemental lighting is provided by
1000 W sodium vapor lamps. After the V10 stage, the ear shoots of
the maize plant, which are ready for fertilization, are enclosed in
a paper bag before silk emergent to withhold the pollen. The ear
shoots are pollinated and 22 days after pollination, the ears are
pulled out and then the kernels are plucked out of the ears. Each
kernel is then dissected into the embryo and the endosperm and the
alurone layer is removed. After dissection, the endosperms are
immediately frozen in liquid nitrogen and then stored at
-80.degree. C. until RNA preparation.
[0414] The CMz030 (Lib143) cDNA library is generated from maize
seedling tissue two days post germination. Seeds are planted on a
moist filter paper on a covered try that is keep in the dark until
germination. The trays are then moved to the bench top at 15 hr
daytime/9 hr nighttime cycles for 2 days post-germination. The day
time temperature is 80.degree. F. and the nighttime temperature is
70.degree. F. Tissue is collected when the seedlings are 2 days
old. At this stage, the colehrhiza has pushed through the seed coat
and the primary root (the radicle) is just piercing the colehrhiza
and is barely visible. The seedlings are placed at 42.degree. C.
for 1 hour. Following the heat shock treatment, the seedlings are
immersed in liquid nitrogen and crushed. The harvested tissue is
stored at -80.degree. until RNA preparation.
[0415] The CMz031 (Lib148) cDNA library is generated from maize
pollen tissue at the V10+ plant development stage. Seeds are
planted at a depth of approximately 3 cm into 2-3 inch peat pots
containing Metro 200 growing medium. After 2-3 weeks growth they
are transplanted into 10 inch pots containing the same growing
medium. Plants are watered daily before transplantation and three
times a week after transplantation. Peters 15-16-17 fertilizer is
applied three times per week after transplanting at a strength of
150 ppm N. Two to three times during the lifetime of the plant,
from transplanting to flowering, a total of 900 mg Fe is added to
each pot. Maize plants are grown in the greenhouse in 15 hr day/9
hr night cycles. The daytime temperature is approximately
80.degree. F. and the nighttime temperature is approximately
70.degree. F. Supplemental lighting is provided by 1000 W sodium
vapor lamps. Tissue is collected from V10+ stage plants. The ear
shoots, which are ready for fertilization, are enclosed in a paper
bag to withhold pollen. Twenty-one days after pollination, prior to
removing the ears, the paper bag is shaken to collect the mature
pollen. The mature pollen is immediately frozen in liquid nitrogen
containers and the pollen is crushed. The harvested tissue is then
stored at -80.degree. C. until RNA preparation.
[0416] The CMz033 (Lib 189) cDNA library is generated from maize
pooled leaf tissue. Samples are harvested from open pollinated
plants. Tissue is collected from maize leaves at the anthesis
stage. The leaves are collect from 10-12 plants and frozen in
liquid nitrogen. The harvested tissue is then stored at -80.degree.
C. until RNA preparation.
[0417] The CMz034 (Lib3060) cDNA library is generated from maize
mature tissue at 40 days post pollination plant development stage.
Seeds are planted at a depth of approximately 3 cm into 2-3 inch
peat pots containing Metro 200 growing medium. After 2-3 weeks
growth they are transplanted into 10 inch pots containing the same
growing medium. Plants are watered daily before transplantation and
three times a week after transplantation. Peters 15-16-17
fertilizer is applied three times per week after transplanting at a
strength of 150 ppm N. Two to three times during the lifetime of
the plant, from transplanting to flowering, a total of 900 mg Fe is
added to each pot. Maize plants are grown in the greenhouse in 15
hr day/9 hr night cycles. The daytime temperature is approximately
80.degree. F. and the nighttime temperature is approximately
70.degree. F. Supplemental lighting is provided by 1000 W sodium
vapor lamps. Tissue is collected from leaves located two leaves
below the ear leaf. This sample represents those genes expressed
during onset and early stages of leaf senescence. The leaves are
pooled and immediately transferred to liquid nitrogen. The
harvested tissue is then stored at -80.degree. C. until RNA
preparation.
[0418] The CMz036 (Lib3062) cDNA library is generated from maize
husk tissue at the 8 week old plant development stage. Seeds are
planted at a depth of approximately 3 cm into 2-3 inch peat pots
containing Metro 200 growing medium. After 2-3 weeks growth they
are transplanted into 10 inch pots containing the same growing
medium. Plants are watered daily before transplantation and three
times a week after transplantation. Peters 15-16-17 fertilizer is
applied three times per week after transplanting at a strength of
150 ppm N. Two to three times during the lifetime of the plant,
from transplanting to flowering, a total of 900 mg Fe is added to
each pot. Maize plants are grown in the greenhouse in 15 hr day/9
hr night cycles. The daytime temperature is approximately
80.degree. F. and the nighttime temperature is approximately
70.degree. F. Supplemental lighting is provided by 1000 W sodium
vapor lamps. Tissue is collected from 8 week old plants. The husk
is separated from the ear and immediately transferred to liquid
nitrogen containers. The harvested tissue is then stored at
-80.degree. C. until RNA preparation.
[0419] The CMz037 (Lib3059) cDNA library is generated from maize
pooled kernal at 12-15 days after pollienation plant development
stage. Sample were collected from field grown material. Whole
kemals from hand pollinated (control pollination) are harvested as
whole ears and immediately frozen on dry ice. Kernels from 10-12
ears were pooled and ground together in liquid nitrogen. The
harvested tissue is then stored at -80.degree. C. until RNA
preparation.
[0420] The CMz039 (Lib3066) cDNA library is generated from maize
immature anther tissue at the 7 week old immature tassel stage.
Seeds are planted at a depth of approximately 3 cm into 2-3 inch
peat pots containing Metro 200 growing medium. After 2-3 weeks
growth they are transplanted into 10 inch pots containing the same
growing medium. Plants are watered daily before transplantation and
three times a week after transplantation. Peters 15-16-17
fertilizer is applied three times per week after transplanting at a
strength of 150 ppm N. Two to three times during the lifetime of
the plant, from transplanting to flowering, a total of 900 mg Fe is
added to each pot. Maize plants are grown in the greenhouse in 15
hr day/9 hr night cycles. The daytime temperature is approximately
80.degree. F. and the nighttime temperature is approximately
70.degree. F. Supplemental lighting is provided by 1000 W sodium
vapor lamps. Tissue is collected when the maize plant is at the 7
week old immature tassel stage. At this stage, prior to anthesis,
the immature anthers are green and enclosed in the staminate
spikelet. The developing anthers are dissected away from the 7 week
old immature tassel and immediately frozen in liquid nitrogen. The
harvested tissue is then stored at -80.degree. C. until RNA
preparation.
[0421] The CMz040 (Lib3067) cDNA library is generated from maize
kernel tissue at the V10+ plant development stage. Seeds are
planted at a depth of approximately 3 cm into 2-3 inch peat pots
containing Metro 200 growing medium. After 2-3 weeks growth they
are transplanted into 10 inch pots containing the same growing
medium. Plants are watered daily before transplantation and three
times a week after transplantation. Peters 15-16-17 fertilizer is
applied three times per week after transplanting at a strength of
150 ppm N. Two to three times during the lifetime of the plant,
from transplanting to flowering, a total of 900 mg Fe is added to
each pot. Maize plants are grown in the greenhouse in 15 hr day/9
hr night cycles. The daytime temperature is approximately
80.degree. F. and the nighttime temperature is approximately
70.degree. F. Supplemental lighting is provided by 1000 W sodium
vapor lamps. Tissue is collected from V10+ stage plants. The ear
shoots, which are ready for fertilization, are enclosed in a paper
bag before silk emergence to withhold pollen. Five to eight days
after controlled pollination. The ears are pulled and the kernels
removed. The kernels are immediately frozen in liquid nitrogen. The
harvested kernels tissue is then stored at -80.degree. C. until RNA
preparation. This sample represents gene expressed in early kernel
development, during periods of cell division, amyloplast biogenesis
and early carbon flow across the material to filial tissue.
[0422] The CMz041 (Lib3068) cDNA library is generated from maize
pollen germinating silk tissue at the V10+ plant development stage.
Seeds are planted at a depth of approximately 3 cm into 2-3 inch
peat pots containing Metro 200 growing medium. After 2-3 weeks
growth they are transplanted into 10 inch pots containing the same
growing medium. Plants are watered daily before transplantation and
three times a week after transplantation. Peters 15-16-17
fertilizer is applied three times per week after transplanting at a
strength of 150 ppm N. Two to three times during the lifetime of
the plant, from transplanting to flowering, a total of 900 mg Fe is
added to each pot. Maize plants are grown in the greenhouse in 15
hr day/9 hr night cycles. The daytime temperature is approximately
80.degree. F. and the nighttime temperature is approximately
70.degree. F. Supplemental lighting is provided by 1000 W sodium
vapor lamps. Tissue is collected from V10+ stage plants when the
ear shoots are ready for fertilization at the silk emergence stage.
The emerging silks are pollinated with an excess of pollen under
controlled pollination conditions in the green house. Eighteen
hours after pollination the silks are removed from the ears and
immediately frozen in liquid nitrogen containers. This sample
represents genes expressed in both pollen and silk tissue early in
pollination. The harvested tissue is then stored at -80.degree. C.
until RNA preparation.
[0423] The CMz042 (Lib3069) cDNA library is generated from maize
ear tissue excessively pollinated at the V10+ plant development
stage. Seeds are planted at a depth of approximately 3 cm into 2-3
inch peat pots containing Metro 200 growing medium. After 2-3 weeks
growth they are transplanted into 10 inch pots containing the same
growing medium. Plants are watered daily before transplantation and
three times a week after transplantation. Peters 15-16-17
fertilizer is applied three times per week after transplanting at a
strength of 150 ppm N. Two to three times during the lifetime of
the plant, from transplanting to flowering, a total of 900 mg Fe is
added to each pot. Maize plants are grown in the greenhouse in 15
hr day/9 hr night cycles. The daytime temperature is approximately
80.degree. F. and the nighttime temperature is approximately
70.degree. F. Supplemental lighting is provided by 1000 W sodium
vapor lamps. Tissue is collected from V10+ stage plants and the ear
shoots which are ready for fertilization are at the silk emergence
stage. The immature ears are pollinated with an excess of pollen
under controlled pollination conditions. Eighteen hours
post-pollination, the ears are removed and immediately transferred
to liquid nitrogen containers. The harvested tissue is then stored
at -80.degree. C. until RNA preparation.
[0424] The CMz044 (Lib3075) cDNA library is generated from maize
microspore tissue at the V10+ plant development stage. Seeds are
planted at a depth of approximately 3 cm into 2-3 inch peat pots
containing Metro 200 growing medium. After 2-3 weeks growth they
are transplanted into 10 inch pots containing the same growing
medium. Plants are watered daily before transplantation and three
times a week after transplantation. Peters 15-16-17 fertilizer is
applied three times per week after transplanting at a strength of
150 ppm N. Two to three times during the lifetime of the plant,
from transplanting to flowering, a total of 900 mg Fe is added to
each pot. Maize plants are grown in the greenhouse in 15 hr day/9
hr night cycles. The daytime temperature is approximately
80.degree. F. and the nighttime temperature is approximately
70.degree. F. Supplemental lighting is provided by 1000 W sodium
vapor lamps. Tissue is collected from immature anthers from 7 week
old tassels. The immature anthers are first dissected from the 7
week old tassel with a scalpel on a glass slide covered with water.
The microspores (immature pollen) are released into the water and
are recovered by centrifugation. The microspore suspension is
immediately frozen in liquid nitrogen. The harvested tissue is then
stored at -80.degree. C. until RNA preparation.
[0425] The CMz045 (Lib3076) cDNA library is generated from maize
immature ear megaspore tissue. Seeds are planted at a depth of
approximately 3 cm into 2-3 inch peat pots containing Metro 200
growing medium. After 2-3 weeks growth they are transplanted into
10 inch pots containing the same growing medium. Plants are watered
daily before transplantation and three times a week after
transplantation. Peters 15-16-17 fertilizer is applied three times
per week after transplanting at a strength of 150 ppm N. Two to
three times during the lifetime of the plant, from transplanting to
flowering, a total of 900 mg Fe is added to each pot. Maize plants
are grown in the greenhouse in 15 hr day/9 hr night cycles. The
daytime temperature is approximately 80.degree. F. and the
nighttime temperature is approximately 70.degree. F. Supplemental
lighting is provided by 1000 W sodium vapor lamps. Tissue is
collected from immature ear (megaspore) obtained from 7 week old
plants. The immature ears are harvested from the 7 week old plants
and are approximately 2.5 to 3 cm in length. The kernels are
removed from the cob immediately frozen in liquid nitrogen. The
harvested tissue is then stored at -80.degree. C. until RNA
preparation.
[0426] The CMz047 (Lib3078) cDNA library is generated from maize
CO.sub.2 treated high-exposure shoot tissue at the V10+ plant
development stage. RX601 maize seeds are sterilized for minute with
a 10% clorox solution. The seeds are rolled in germination paper,
and germinated in 0.5 mM calcium sulfate solution for two days ate
30.degree. C. The seedlings are planted at a depth of approximately
3 cm into 2-3 inch peat pots containing Metro 200 growing medium at
a rate of 2-3 seedlings per pot. Twenty pots are placed into a high
CO.sub.2 environment (approximately 1000 ppm CO.sub.2). Twenty
plants were grown under ambient greenhouse CO.sub.2 (approximately
450 ppm CO.sub.2). Plants are watered daily before transplantation
and three times a week after transplantation. Peters 20-20-20
fertilizer is also lightly applied. Maize plants are grown in the
greenhouse in 15 hr day/9 hr night cycles. The daytime temperature
is approximately 80.degree. F. and the nighttime temperature is
approximately 70.degree. F. Supplemental lighting is provided by
1000 W sodium vapor lamps. At ten days post planting, the shoots
from both atmosphere are frozen in liquid nitrogen and lightly
ground. The roots are washed in deionized water to remove the
support media and the tissue is immediately transferred to liquid
nitrogen containers. The harvested tissue is then stored at
-80.degree. C. until RNA preparation.
[0427] The CMz048 (Lib3079) cDNA library is generated from maize
basal endosperm transfer layer tissue at the V10+ plant development
stage. Seeds are planted at a depth of approximately 3 cm into 2-3
inch peat pots containing Metro 200 growing medium. After 2-3 weeks
growth they are transplanted into 10 inch pots containing the same
growing medium. Plants are watered daily before transplantation and
three times a week after transplantation. Peters 15-16-17
fertilizer is applied three times per week after transplanting at a
strength of 150 ppm N. Two to three times during the lifetime of
the plant, from transplanting to flowering, a total of 900 mg Fe is
added to each pot. Maize plants are grown in the greenhouse in 15
hr day/9 hr night cycles. The daytime temperature is approximately
80.degree. F. and the nighttime temperature is approximately
70.degree. F. Supplemental lighting is provided by 1000 W sodium
vapor lamps. Tissue is collected from V10+ maize plants. The ear
shoots, which are ready for fertilization, are enclosed in a paper
bag prior to silk emergence, to withhold the pollen. Kernels are
harvested at 12 days post-pollination and placed on wet ice for
dissection. The kernels are cross sectioned laterally, dissecting
just aboVe the pedicel region, including 1-2 mm of the lower
endosperm and the basal endosperm transfer region. The pedicel and
lower endosperm region containing the basal endosperm transfer
layer is pooled and immediately frozen in liquid nitrogen. The
harvested tissue is then stored at -80.degree. C. until RNA
preparation.
[0428] The CMz049(Lib3088) cDNA library is generated from maize
immature anther tissue at the 7 week old immature tassel stage.
Seeds are planted at a depth of approximately 3 cm into 2-3 inch
peat pots containing Metro 200 growing medium. After 2-3 weeks
growth they are transplanted into 10 inch pots containing the same
growing medium. Plants are watered daily before transplantation and
three times a week after transplantation. Peters 15-16-17
fertilizer is applied three times per week after transplanting at a
strength of 150 ppm N. Two to three times during the lifetime of
the plant, from transplanting to flowering, a total of 900 mg Fe is
added to each pot. Maize plants are grown in the greenhouse in 15
hr day/9 hr night cycles. The daytime temperature is approximately
80.degree. F. and the nighttime temperature is approximately
70.degree. F. Supplemental lighting is provided by 1000 W sodium
vapor lamps. Tissue is collected when the maize plant is at the 7
week old immature tassel stage. At this stage, prior to anthesis,
the immature anthers are green and enclosed in the staminate
spikelet. The developing anthers are dissected away from the 7 week
old immature tassel and immediately transferred to liquid nitrogen
container. The harvested tissue is then stored at -80.degree. C.
until RNA preparation.
[0429] The CMz050 (Lib3114) cDNA library is generated from maize
silk tissue at the V10+ plant development stage. Seeds are planted
at a depth of approximately 3 cm into 2-3 inch peat pots containing
Metro 200 growing medium. After 2-3 weeks growth they are
transplanted into 10 inch pots containing the same growing medium.
Plants are watered daily before transplantation and three times a
week after transplantation. Peters 15-16-17 fertilizer is applied
three times per week after transplanting at a strength of 150 ppm
N. Two to three times during the lifetime of the plant, from
transplanting to flowering, a total of 900 mg Fe is added to each
pot. Maize plants are grown in the greenhouse in 15 hr day/9 hr
night cycles. The daytime temperature is approximately 80.degree.
F. and the nighttime temperature is approximately 70.degree. F.
Supplemental lighting is provided by 1000 W sodium vapor lamps.
Tissue is collected when the maize plant is beyond the 10-leaf
development stage and the ear shoots are approximately 15-20 cm in
length. The ears are pulled and silks are separated from the ears
and immediately transferred to liquid nitrogen containers. The
harvested tissue is then stored at -80.degree. C. until RNA
preparation.
[0430] The SOYMON001 cDNA library is generated from soybean
cultivar Asgrow 3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.)
total leaf tissue at the V4 plant development stage. Leaf tissue
from 38, field grown V4 stage plants is harvested from the 4.sup.th
node. Leaf tissue is removed from the plants and immediately frozen
in dry-ice. The harvested tissue is then stored at -80.degree. C.
until RNA preparation.
[0431] The SOYMON002 cDNA library is generated from soybean
cultivar Asgrow 3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.)
root tissue at the V4 plant development stage. Root tissue from 76,
field grown V4 stage plants is harvested. The root systems is cut
from the soybean plant and washed with water to free it from the
soil and immediately frozen in dry-ice. The harvested tissue is
then stored at -80.degree. C. until RNA preparation.
[0432] The SOYMON003 cDNA library is generated from soybean
cultivar Asgrow 3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.)
seedling hypocotyl axis tissue harvested 2 day post-imbibition.
Seeds are planted at a depth of approximately 2 cm into 2-3 inch
peat pots containing Metromix 350 medium. Trays are placed in an
environmental chamber and grown at 12 hr daytime/12 hr nighttime
cycles. The daytime temperature is approximately 29.degree. C. and
the nighttime temperature approximately 24.degree. C. Soil is
checked and watered daily to maintain even moisture conditions.
Tissue is collected 2 days after the start of imbibition. The 2
days after imbibition samples are separated into 3 collections
after removal of any adhering seed coat. At the 2 day stage, the
hypocotyl axis is emerging from the soil. A few seedlings have
cracked the soil surface and exhibited slight greening of the
exposed cotyledons. The seedlings are washed in water to remove
soil, hypocotyl axis harvested and immediately frozen in liquid
nitrogen. The harvested tissue is then stored at -80.degree. C.
until RNA preparation.
[0433] The SOYMON004 cDNA library is generated from soybean
cultivar Asgrow 3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.)
seedling cotyledon tissue harvested 2 day post-imbibition. Seeds
are planted at a depth of approximately 2 cm into 2-3 inch peat
pots containing Metromix 350 medium. Trays are placed in an
environmental chamber and grown at 12 hr daytime/12 hr nighttime
cycles. The daytime temperature is approximately 29.degree. C. and
the nighttime temperature approximately 24.degree. C. Soil is
checked and watered daily to maintain even moisture conditions.
Tissue is collected 2 days after the start of imbibition. The 2
days after imbibition samples are separated into 3 collections
after removal of any adhering seed coat. At the 2 day stage, the
hypocotyl axis is emerging from the soil. A few seedlings have
cracked the soil surface and exhibited slight greening of the
exposed cotyledons. The seedlings are washed in water to remove
soil, hypocotyl axis harvested and immediately frozen in liquid
nitrogen. The harvested tissue is then stored at -80.degree. C.
until RNA preparation.
[0434] The SOYMON005 cDNA library is generated from soybean
cultivar Asgrow 3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.)
seedling hypocotyl axis tissue harvested 6 hour post-imbibition.
Seeds are planted at a depth of approximately 2 cm into 2-3 inch
peat pots containing Metromix 350 medium. Trays are placed in an
environmental chamber and grown at 12 hr daytime/12 hr nighttime
cycles. The daytime temperature is approximately 29.degree. C. and
the nighttime temperature approximately 24.degree. C. Soil is
checked and watered daily to maintain even moisture conditions.
Tissue is collected 6 hours after the start of imbibition. The 6
hours after imbibition samples are separated into 3 collections
after removal of any adhering seed coat. The 6 hours after
imbibition sample is collected over the course of approximately 2
hours starting at 6 hours post imbibition. At the 6 hours after
imbibition stage, not all cotyledons have become fully hydrated and
germination, or radicle protrusion, has not occurred. The seedlings
are washed in water to remove soil, hypocotyl axis harvested and
immediately frozen in liquid nitrogen. The harvested tissue is then
stored at -80.degree. C. until RNA preparation.
[0435] The SOYMON006 cDNA library is generated from soybean
cultivar Asgrow 3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.)
seedling cotyledons tissue harvest 6 hour post-imbibition. Seeds
are planted at a depth of approximately 2 cm into 2-3 inch peat
pots containing Metromix 350 medium. Trays are placed in an
environmental chamber and grown at 12 hr daytime/12 hr nightime
cycles. The daytime temperature is approximately 29.degree. C. and
the nighttime temperature approximately 24.degree. C. Soil is
checked and watered daily to maintain even moisture conditions.
Tissue is collected 6 hours after imbibition. The 6 hours after
imbibition samples are separated into 3 collections after removal
of any adhering seed coat. The 6 hours after imbibition sample is
collected over the course of approximately 2 hours starting at 6
hours post-imbibition. At the 6 hours after imbibition, not all
cotyledons have become fully hydrated and germination or radicle
protrusion, have not occurred. The seedlings are washed in water to
remove soil, cotyledon harvested and immediately frozen in liquid
nitrogen. The harvested tissue is then stored at -80.degree. C.
until RNA preparation.
[0436] The SOYMON007 cDNA library is generated from soybean
cultivar Asgrow 3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.)
seed tissue harvested 25 and 35 days post-flowering. Seed pods from
field grown plants are harvested 25 and 35 days after flowering and
the seeds extracted from the pods. Approximately 4.4 g and 19.3 g
of seeds are harvested from the respective seed pods and
immediately frozen in dry ice. The harvested tissue is then stored
at -80.degree. C. until RNA preparation.
[0437] The SOYMON008 cDNA library is generated from soybean
cultivar Asgrow 3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.)
leaf tissue harvested from 25 and 35 days post-flowering plants.
Total leaf tissue is harvested from field grovin plants.
Approximately 19 g and 29 g of leaves are harvested from the fourth
node of the plant 25 and 35 days post-flowering and immediately
frozen in dry ice. The harvested tissue is then stored at
-80.degree. C. until RNA preparation.
[0438] The SOYMON009 cDNA library is generated from soybean
cutlivar C1944 (USDA Soybean Germplasm Collection, Urbana, Ill.
U.S.A.) pod and seed tissue harvested 15 days post-flowering. Pods
from field grown plants are harvested 15 days post-flowering.
Approximately 3 g of pod tissue is harvested and immediately frozen
in dry-ice. The harvested tissue is then stored at -80.degree. C.
until RNA preparation.
[0439] The SOYMON010 cDNA library is generated from soybean
cultivar C1944 (USDA Soybean Germplasm Collection, Urbana, Ill.
U.S.A.) seed tissue harvested 40 days post-flowering. Pods from
field grown plants are harvested 40 days post-flowering. Pods and
seeds are separated, approximately 19 g of seed tissue is harvested
and immediately frozen in dry-ice. The harvested tissue is then
stored at -80.degree. C. until RNA preparation.
[0440] The SOYMON011 cDNA library is generated from soybean
cultivars Cristalina (USDA Soybean Germplasm Collection, Urbana,
Ill. U.S.A.) and FT108 (Monsoy, Brazil) (tropical germ plasma) leaf
tissue. Leaves are harvested from plants grown in an environmental
chamber under 12 hr daytime/12 hr nighttime cycles. The daytime
temperature is approximately 29.degree. C. and the nighttime
temperature approximately 24.degree. C. Soil is checked and watered
daily to maintain even moisture conditions. Approximately 30 g of
leaves are harvested from the 4.sup.th node of each of the
Cristalina and FT108 cultivars and immediately frozen in dry ice.
The harvested tissue is then stored at -80.degree. C. until RNA
preparation.
[0441] The SOYMON012 cDNA library is generated from soybean
cultivar Asgrow 3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.)
leaf tissue. Leaves from field grown plants are harvested from the
fourth node 15 days post-flowering. Approximately 12 g of leaves
are harvested and immediately frozen in dry ice. The harvested
tissue is then stored at -80.degree. C. until RNA preparation.
[0442] The SOYMON013 cDNA library is generated from soybean
cultivar Asgrow 3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.)
root and nodule tissue. Approximately, 28 g of root tissue from
field grown plants is harvested 15 days post-flowering. The root
system is cut from the soybean plant, washed with water to free it
from the soil and immediately frozen in dry-ice. The harvested
tissue is then stored at -80.degree. C. until RNA preparation.
[0443] The SOYMON014 cDNA library is generated from soybean
cultivar Asgrow 3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.)
seed tissue harvested 25 and 35 days after flowering. Seed pods
from field grown plants are harvested 15 days after flowering and
the seeds extracted from the pods. Approximately 5 g of seeds are
harvested from the respective seed pods and immediately frozen in
dry ice. The harvested tissue is then stored at -80.degree. C.
until RNA preparation.
[0444] The SOYMON015 cDNA is generated from soybean cultivar Asgrow
3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.) seed tissue
harvested 45 and 55 days post-flowering. Seed pods from field grown
plants are harvested 45 and 55 days after flowering and the seeds
extracted from the pods. Approximately 19 g and 31 g of seeds are
harvested from the respective seed pods and immediately frozen in
dry ice. The harvested tissue is then stored at -80.degree. C.
until RNA preparation.
[0445] The SOYMON016 cDNA library is generated from soybean
cultivar Asgrow 3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.)
root tissue. Approximately, 61 g and 38 g of root tissue from field
grown plants is harvested 25 and 35 days post-flowering is
harvested. The root system is cut from the soybean plant and washed
with water to free it from the soil. The tissue is placed in 14 ml
polystyrene tubes and immediately frozen in dry-ice. The harvested
tissue is then stored at -80.degree. C. until RNA preparation.
[0446] The SOYMON017 cDNA library is generated from soybean
cultivar Asgrow 3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.)
root tissue. Approximately 28 g of root tissue from field grown
plants is harvested 45 and 55 days post-flowering. The root system
is cut from the soybean plant, washed with water to free it from
the soil and immediately frozen in dry-ice. The harvested tissue is
then stored at -80.degree. C. until RNA preparation.
[0447] The SOYMON018 cDNA is generated from soybean cultivar Asgrow
3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.) leaf tissue
harvested 45 and 55 days post-flowering. Leaves from field grown
plants are harvested 45 and 55 days after flowering from the fourth
node. Approximately 27 g and 33 g of seeds are harvested from the
respective seed pods and immediately frozen in dry ice. The
harvested tissue is then stored at -80.degree. C. until RNA
preparation.
[0448] The SOYMON019 cDNA library is generated from soybean
cultivars Cristalina (USDA Soybean Germplasm Collection, Urbana,
Ill. U.S.A.) and FT108 (Monsoy, Brazil) (tropical germ plasma) root
tissue. Roots are harvested from plants grown in an environmental
chamber under 12 hr daytime/12 hr nighttime cycles. The daytime
temperature is approximately 29.degree. C. and the nighttime
temperature approximately 24.degree. C. Soil is checked and watered
daily to maintain even moisture conditions. Approximately 50 g and
56 g of roots are harvested from each of the Cristalina and FT108
cultivars and immediately frozen in dry ice. The harvested tissue
is then stored at -80.degree. C. until RNA preparation.
[0449] The SOYMON020 cDNA is generated from soybean cultivar Asgrow
3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.) seed tissue
harvested 65 and 75 days post-flowering. Seed pods from field grown
plants are harvested 45 and 55 days after flowering and the seeds
extracted from the pods. Approximately 14 g and 31 g of seeds are
harvested from the respective seed pods and immediately frozen in
dry ice. The harvested tissue is then stored at -80.degree. C.
until RNA preparation.
[0450] The SOYMON021 cDNA library is generated from Soybean Cyst
Nematode-resistant soybean cultivar Hartwig (USDA Soybean Germplasm
Collection, Urbana, Ill. U.S.A.) root tissue. Plants are grown in
tissue culture at room temperature. At approximately 6 weeks
post-germination, the plants are exposed to sterilized Soybean Cyst
Nematode eggs. Infection is then allowed to progress for 10 days.
After the 10 day infection process, the tissue is harvested. Agar
from the culture medium and nematodes are removed and the root
tissue is immediately frozen in dry ice. The harvested tissue is
then stored at -80.degree. C. until RNA preparation.
[0451] The SOYMON022 (Lib3030) cDNA library is generated from
soybean cultivar Asgrow 3244 (Asgrow Seed Company, Des Moines, Iowa
U.S.A.) partially opened flower tissue. Partially to fully opened
flower tissue is harvested from plants grown in an environmental
chamber under 12 hr daytime/12 hr nighttime cycles. The daytime
temperature is approximately 29.degree. C. and the nighttime
temperature approximately 24.degree. C. Soil is checked and watered
daily to maintain even moisture conditions. A total of 3 g of
flower tissue is harvested and immediately frozen in dry ice. The
harvested tissue is then stored at -80.degree. C. until RNA
preparation.
[0452] The SOYMON023 cDNA library is generated from soybean
genotype BW211S Null (Tohoku University, Morioka, Japan) seed
tissue harvested 15 and 40 days post-flowering. Seed pods from
field grown plants are harvested 15 and 40 days post-flowering and
the seeds extracted from the pods. Approximately 0.7 g and 14.2 g
of seeds are harvested from the respective seed pods and
immediately frozen in dry ice. The harvested tissue is then stored
at -80.degree. C. until RNA preparation.
[0453] The SOYMON024 cDNA library is generated from soybean
cultivar Asgrow 3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.)
internode-2 tissue harvested 18 days post-imbibition. Seeds are
planted at a depth of approximately 2 cm into 2-3 inch peat pots
containing Metromix 350 medium. The plants are grown in a
greenhouse for 18 days after the start of imbibition at ambient
temperature. Soil is checked and watered daily to maintain even
moisture conditions. Stem tissue is harvested 18 days after the
start of imbibition. The samples are divided into hypocotyl and
internodes 1 through 5. The fifth internode contains some leaf bud
material. Approximately 3 g of each sample is harvested and
immediately frozen in dry ice. The harvested tissue is then stored
at -80.degree. C. until RNA preparation.
[0454] The SOYMON025 cDNA library is generated from soybean
cultivar Asgrow 3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.)
leaf tissue harvested 65 days post-flowering. Leaves are harvested
from the fourth node of field grown plants 65 days post-flowering.
Approximately 18.4 g of leaf tissue is harvested and immediately
frozen in dry ice. The harvested tissue is then stored at
-80.degree. C. until RNA preparation.
[0455] SOYMON026 cDNA library is generated from soybean cultivar
Asgrow 3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.) root
tissue harvested 65 and 75 days post-flowering. Approximately 27 g
and 40 g of root tissue from field grown plants is harvested 65 and
75 days post-flowering. The root system is cut from the soybean
plant, washed with water to free it from the soil and immediately
frozen in dry-ice. The harvested tissue is then stored at
-80.degree. C. until RNA preparation.
[0456] The SOYMON027 cDNA library is generated from soybean
cultivar Asgrow 3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.)
seed tissue harvested 25 days post-flowering. Seed pods from field
grown plants are harvested 25 days post-flowering and the seeds
extracted from the pods. Approximately 17 g of seeds are harvested
from the seed pods and immediately frozen in dry ice. The harvested
tissue is then stored at -80.degree. C. until RNA preparation.
[0457] The SOYMON028 cDNA library is generated from soybean
cultivar Asgrow 3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.)
drought-stressed root tissue. The plants are grown in an
environmental chamber under 12 hr daytime/12 hr nighttime cycles.
The daytime temperature is approximately 29.degree. C. and the
nighttime temperature 24.degree. C. Soil is checked and watered
daily to maintain even moisture conditions. At the R3 stage of
development, water is withheld from half of the plant collection
(drought stressed population). After 3 days, half of the plants
from the drought stressed condition and half of the plants from the
control population are harvested. After another 3 days (6 days post
drought induction) the remaining plants are harvested. A total of
27 g and 40 g of root tissue is harvested and immediately frozen in
dry ice. The harvested tissue is then stored at -80.degree. C.
until RNA preparation.
[0458] The SOYMON029 cDNA library is generated from Soybean Cyst
Nematode-resistant soybean cultivar PI07354 (USDA Soybean Germplasm
Collection, Urbana, Ill. U.S.A.) root tissue. Late fall to early
winter greenhouse grown plants are exposed to Soybean Cyst Nematode
eggs. At 10 days post-infection, the plants are uprooted, rinsed
briefly and the roots frozen in liquid nitrogen. Approximately 20
grams of root tissue is harvested from the infected plants. The
harvested tissue is then stored at -80.degree. C. until RNA
preparation.
[0459] The SOYMON030 cDNA library is generated from soybean
cultivar Asgrow 3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.)
flower bud tissue. Seeds are planted at a depth of approximately 2
cm into 2-3 inch peat pots containing Metromix 350 medium and the
plants are grown in an environmental chamber under 12 hr daytime/12
hr nighttime cycles. The daytime temperature is approximately
29.degree. C. and the nighttime temperature approximately
24.degree. C. Soil is checked and watered daily to maintain even
moisture conditions. Flower buds are removed from the plant at the
pedicel. A total of 100 mg of flower buds are harvested and
immediately frozen in liquid nitrogen. The harvested tissue is then
stored at -80.degree. C. until RNA preparation.
[0460] The SOYMON031 cDNA library is generated from soybean
cultivar Asgrow 3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.)
carpel and stamen tissue. Seeds are planted at a depth of
approximately 2 cm into 2-3 inch peat pots containing Metromix 350
medium and the plants are grown in an environmental chamber under
12 hr daytime/12 hr nighttime cycles. The daytime temperature is
approximately 29.degree. C. and the nighttime temperature
approximately 24.degree. C. Soil is checked and watered daily to
maintain even moisture conditions. Flower buds are removed from the
plant at the pedicel. Flowers are dissected to separate petals,
sepals and reproductive structures (carpels and stamens). A total
of 300 mg of carpel and stamen tissue are harvested and immediately
frozen in liquid nitrogen. The harvested tissue is then stored at
-80.degree. C. until RNA preparation.
[0461] The SOYMON032 cDNA library is prepared from the Asgrow
cultivar A4922 (Asgrow Seed Company, Des Moines, Iowa U.S.A.)
rehydrated dry soybean seed meristem tissue. Surface sterilized
seeds are germinated in liquid media for 24 hours. The seed axis is
then excised from the barely germinating seed, placed on tissue
culture media and incubated overnight at 20.degree. C. in the dark.
The supportive tissue is removed from the explant prior to harvest.
Approximately 570 mg of tissue is harvested and frozen in liquid
nitrogen. The harvested tissue is then stored at -80.degree. C.
until RNA preparation.
[0462] The SOYMON033 cDNA library is generated from soybean
cultivar Asgrow 3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.)
heat-shocked seedling tissue without cotyledons. Seeds are imbibed
and germinated in vermiculite for 2 days under constant
illumination. After 48 hours, the seedlings are transferred to an
incubator set at 40.degree. C. under constant illumination. After
30, 60 and 180 minutes seedlings are harvested and dissected. A
portion of the seedling consisting of the root, hypocotyl and
apical hook is frozen in liquid nitrogen and stored at -80.degree.
C. The seedlings after 2 days of imbibition are beginning to emerge
from the vermiculite surface. The apical hooks are dark green in
appearance. Total RNA and poly A.sup.+ RNA is prepared from equal
amounts of pooled tissue.
[0463] The SOYMON034 cDNA library is generated from soybean
cultivar Asgrow 3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.)
cold-shocked seedling tissue without cotyledons. Seeds are imbibed
and germinated in vermiculite for 2 days under constant
illumination. After 48 hours, the seedlings are transferred to a
cold room set at 5.degree. C. under constant illumination. After
30, 60 and 180 minutes seedlings are harvested and dissected. A
portion of the seedling consisting of the root, hypocotyl and
apical hook is frozen in liquid nitrogen and stored at -80.degree.
C. The seedlings after 2 days of imbibition are beginning to emerge
from the vermiculite surface. The apical hooks are dark green in
appearance.
[0464] The SOYMON035 cDNA library is generated from soybean
cultivar Asgrow 3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.)
seed coat tissue. Seeds are planted at a depth of approximately 2
cm into 2-3 inch peat pots containing Metromix 350 medium and the
plants are grown in an environmental chamber under 12 hr daytime/12
hr nighttime cycles. The daytime temperature is approximately
29.degree. C. and the nighttime temperature 24.degree. C. Soil is
checked and watered daily to maintain even moisture conditions.
Seeds are harvested from mid to nearly full maturation (seed coats
are not yellowing). The entire embryo proper is removed from the
seed coat sample and the seed coat tissue are harvested and
immediately frozen in liquid nitrogen. The harvested tissue is then
stored at -80.degree. C. until RNA preparation.
[0465] The SOYMON036 cDNA library is generated from soybean
cultivars PI171451, PI227687 and PI229358 (USDA Soybean Germplasm
Collection, Urbana, Ill. U.S.A.) insect challenged leaves. Plants
from each of the three cultivars are grown in screenhouse
conditions. The screenhouse is divided in half and one half of the
screenhouse is infested with soybean looper and the other half
infested with velvetbean caterpillar. A single leaf is taken from
each of the representative plants at 3 different time points, 11
days after infestation, 2 weeks after infestation and 5 weeks after
infestation and immediately frozen in liquid nitrogen. The
harvested tissue is then stored at -80.degree. C. until RNA
preparation. Total RNA and poly A+ RNA is isolated from pooled
tissue consisting of equal quantities of all 18 samples (3
genotypes X 3 sample times X 2 insect genotypes).
[0466] The SOYMON037 cDNA library is generated from soybean
cultivar A3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.)
etiolated axis and radical tissue. Seeds are planted in moist
vermiculite, wrapped and kept at room temperature in complete
darkness until harvest. Etiolated axis and hypocotyl tissue is
harvested at 2, 3 and 4 days post-planting. A total of 1 gram of
each tissue type is harvested at 2, 3 and 4 days after planting and
immediately frozen in liquid nitrogen. The harvested tissue is then
stored at -80.degree. C. until RNA preparation.
[0467] The SOYMON038 cDNA library is generated from soybean variety
Asgrow A3237 (Asgrow Seed Company, Des Moines, Iowa U.S.A.)
rehydrated dry seeds. Explants are prepared for transformation
after germination of surface-sterilized seeds on solid tissue
media. After 6 days, at 28.degree. C. and 18 hours of light per
day, the germinated seeds are cold shocked at 4.degree. C. for 24
hours. Meristemic tissue and part of the hypocotyl is remove and
cotyledon excised. The prepared explant is then wounded for
Agrobacterium infection. The 2 grams of harvested tissue is frozen
in liquid nitrogen and stored at -80.degree. C. until RNA
preparation.
[0468] The Soy51 (LIB3027) cDNA library is prepared from equal
amounts tissue harvested from SOYMON007, SOYMON015 and SOYMON020
prepared tissue. Single stranded and double stranded DNA
representing approximately 1.times.10.sup.6 colony forming units
are isolated using standard protocols. RNA, complementary to the
single stranded DNA, is synthesized using the double stranded DNA
as a template. Biotinylated dATP is incorporated into the RNA
during the synthesis reaction. The single stranded DNA is mixed
with the biotinylated RNA in a 1:10 molar ratio) and allowed to
hybridize. DNA-RNA hybrids are captured on Dynabeads M280
streptavidin (Dynabeads, Dynal Corporation, Lake Success, N.Y.
U.S.A.). The dynabeads with captured hybrids are collected with a
magnet. The non-hybridized single stranded molecules remaining
after hybrid capture are converted to double stranded form and
represent the primary normalized library.
[0469] The Soy52 (LIB3028) cDNA library is generated from
normalized flower DNA. Single stranded DNA representing
approximately 1.times.10.sup.6 colony forming units of SOYMON022
harvested tissue is used as the starting material for
normalization. RNA, complementary to the single stranded DNA, is
synthesized using the double stranded DNA as a template.
Biotinylated dATP is incorporated into the RNA during the synthesis
reaction. The single stranded DNA is mixed with the biotinylated
RNA in a 1:10 molar ratio) and allowed to hybridize. DNA-RNA
hybrids are captured on Dynabeads M280 streptavidin (Dynabeads,
Dynal Corporation, Lake Success, N.Y. U.S.A.). The dynabeads with
captured hybrids are collected with a magnet. The non-hybridized
single stranded molecules remaining after hybrid capture are
converted to double stranded form and represent the primary
normalized library.
[0470] The Soy53 (LIB3039) cDNA library is generated from soybean
cultivar Asgrow 3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.)
seedling shoot apical meristem tissue. Seeds are planted at a depth
of approximately 2 cm into 2-3 inch peat pots containing Metromix
350 medium and the plants are grown in an environmental chamber
under 12 hr daytime/12 hr nighttime cycles. The daytime temperature
is approximately 29.degree. C. and the nighttime temperature
24.degree. C. Soil is checked and watered daily to maintain even
moisture conditions. Apical tissue is harvested from seedling shoot
meristem tissue, 7-8 days after the start of imbibition. The apex
of each seedling is dissected to include the fifth node to the
apical meristem. The fifth node corresponds to the third trifoliate
leaf in the very early stages of development. Stipules completely
envelop the leaf primordia at this time. A total of 200 mg of
apical tissue is harvested and immediately frozen in liquid
nitrogen. The harvested tissue is then stored at -80.degree. C.
until RNA preparation.
[0471] The Soy54 (LIB3040) cDNA library is generated from soybean
cultivar Asgrow 3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.)
heart to torpedo stage embryo tissue. Seeds are planted at a depth
of approximately 2 cm into 2-3 inch peat pots containing Metromix
350 medium and the plants are grown in an environmental chamber
under 12 hr daytime/12 hr nighttime cycles. The daytime temperature
is approximately 29.degree. C. and the nighttime temperature
24.degree. C. Soil is checked and watered daily to maintain even
moisture conditions. Seeds are collected and embryos removed from
surrounding endosperm and maternal tissues. Embryos from globular
to young torpedo stages (by corresponding analogy to Arabidopsis)
are collected with a bias towards the middle of this spectrum.
Embryos which are beginning to show asymmetric development of
cotyledons are considered the upper developmental boundary for the
collection and are excluded. A total of 12 mg embryo tissue is
frozen in liquid nitrogen. The harvested tissue is stored at
-80.degree. C. until RNA preparation.
[0472] Soy55 (LIB3049) cDNA library is generated from soybean
cultivar Asgrow 3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.)
young seed tissue. Seeds are planted at a depth of approximately 2
cm into 2-3 inch peat pots containing Metromix 350 medium and the
plants are grown in an environmental chamber under 12 hr daytime/12
hr nighttime cycles. The daytime temperature is approximately
29.degree. C. and the nighttime temperature 24.degree. C. Soil is
checked and watered daily to maintain even moisture conditions.
Seeds are collected from very young pods (5 to 15 days after
flowering). A total of 100 mg of seeds are harvested and frozen in
liquid nitrogen. The harvested tissue is stored at -80.degree. C.
until RNA preparation.
[0473] Soy56 (LIB3029) cDNA library is prepared from equal amounts
tissue harvested from SOYMON007, SOYMON015 and SOYMON020 prepared
tissue. Single stranded and double stranded DNA representing
approximately 1.times.10.sup.6 colony forming units are isolated
using standard protocols. RNA, complementary to the single stranded
DNA, is synthesized using the double stranded DNA as a template.
Biotinylated dATP is incorporated into the RNA during the synthesis
reaction. The single stranded DNA is mixed with the biotinylated
RNA in a 1:10 molar ratio and allowed to hybridize. DNA-RNA hybrids
are captured on Dynabeads M280 streptavidin (Dynabeads, Dynal
Corporation, Lake Success, N.Y. U.S.A.). The dynabeads with
captured hybrids are collected with a magnet. The non-hybridized
single stranded molecules remaining after hybrid capture are not
converted to double stranded form and represent a non-normalized
seed pool for comparison to Soy51 cDNA libraries.
[0474] TheSoy58 (LIB3050) cDNA library is generated from soybean
cultivar Asgrow 3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.)
drought stressed root tissue subtracted from control root tissue.
Seeds are planted at a depth of approximately 2 cm into 2-3 inch
peat pots containing Metromix 350 medium and the plants are grown
in an environmental chamber under 12 hr daytime/12 hr nighttime
cycles. The daytime temperature is approximately 29.degree. C. and
the nighttime temperature 24.degree. C. Soil is checked and watered
daily to maintain even moisture conditions. At the R3 stage of the
plant drought is induced by withholding water. After 3 and 6 days
root tissue from both drought stressed and control (watered
regularly) plants are collected and frozen in dry-ice. The
harvested tissue is stored at -80.degree. C. until RNA preparation.
For subtraction, target cDNA is made from the drought stressed
tissue total RNA using the SMART cDNA synthesis system from
Clonetech (Clonetech Laboratories, Palo Alto, Calif. U.S.A.).
Driver first strand cDNA is covalently linked to Dynabeads
following a protocol similar to that described in the Dynal
literature (Dynabeads, Dynal Corporation, Lake Success, N.Y.
U.S.A.). The target cDNA is then heat denatured and the second
strand trapped using Dynabeads oligo-dT. The target second strand
cDNA is then hybridized to the driver cDNA in 400 .mu.l
2.times.SSPE for two rounds of hybridization at 65.degree. C. and
20 hours. After each hybridization, the hybridization solution is
removed from the system and the hybridized target cDNA removed from
the driver by heat denaturation in water. After hybridization, the
remaining cDNA is trapped with Dynabeads oligo-dT. The trapped cDNA
is then amplified as in previous PCR based libraries and the
resulting cDNA ligated into the pSPORT vector (Invitrogen, Carlsbad
Calif. U.S.A.).
[0475] The Soy59 (LIB3051) cDNA library is generated from soybean
cultivar Asgrow 3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.)
endosperm tissue. Seeds are germinated on paper towels under
laboratory ambient light conditions. At 8, 10 and 14 hours after
imbibition, the seed coats are harvested. The endosperm consists of
a very thin layer of tissue affixed to the inside of the seed coat.
The seed coat and endosperm are frozen immediately after harvest in
liquid nitrogen. The harvested tissue is stored at -80.degree. C.
until RNA preparation.
[0476] The Soy60 (LIB3072) cDNA library is generated from soybean
cultivar Asgrow 3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.)
drought stressed seed plus pod subtracted from control seed plus
pod tissue. Seeds are planted at a depth of approximately 2 cm into
2-3 inch peat pots containing Metromix 350 medium and the plants
are grown in an environmental chamber under 12 hr daytime/12 hr
nighttime cycles. The daytime temperature is approximately
26.degree. C. and the nighttime temperature 21.degree. C. and 70%
relative humidity. Soil is checked and watered daily to maintain
even moisture conditions. At the R3 stage of the plant drought is
induced by withholding water. After 3 and 6 days seeds and pods
from both drought stressed and control (watered regularly) plants
are collected from the fifth and sixth node and frozen in dry-ice.
The harvested tissue is stored at -80.degree. C. until RNA
preparation. For subtraction, target cDNA is made from the drought
stressed tissue total RNA using the SMART cDNA synthesis system
from Clonetech (Clonetech Laboratories, Palo Alto, Calif. U.S.A.).
Driver first strand cDNA is covalently linked to Dynabeads
following a protocol similar to that described in the Dynal
literature (Dynabeads, Dynal Corporation, Lake Success, N.Y.
U.S.A.). The target cDNA is then heat denatured and the second
strand trapped using Dynabeads oligo-dT. The target second strand
cDNA is then hybridized to the driver cDNA in 400 .mu.l
2.times.SSPE for two rounds of hybridization at 65.degree. C. and
20 hours. After each hybridization, the hybridization solution is
removed from the system and the hybridized target cDNA removed from
the driver by heat denaturation in water. After hybridization, the
remaining cDNA is trapped with Dynabeads oligo-dT. The trapped cDNA
is then amplified as in previous PCR based libraries and the
resulting cDNA ligated into the pSPORT vector (Invitrogen, Carlsbad
Calif. U.S.A.).
[0477] The Soy61 (LIB3073) cDNA library is generated from soybean
cultivar Asgrow 3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.)
jasmonic acid treated seedling subtracted from control tissue.
Seeds are planted at a depth of approximately 2 cm into 2-3 inch
peat pots containing Metromix 350 medium and the plants are grown
in a greenhouse. The daytime temperature is approximately
29.4.degree. C. and the nighttime temperature 20.degree. C. Soil is
checked and watered daily to maintain even moisture conditions. At
9 days post planting, the plantlets are sprayed with either control
buffer of 0.1% Tween-20 or jasmonic acid (Sigma J-2500, Sigma, St.
Loius, Mo. U.S.A.) at 1 mg/ml in 0.1% Tween-20. Plants are sprayed
until runoff and the soil and the stem is socked with the spraying
solution. At 18 hours post application of jasmonic acid, the
soybean plantlets appear growth retarded. After 18 hours, 24 hours
and 48 hours post treatment, the cotyledons are removed and the
remaining leaf and stem tissue above the soil is harvested and
frozen in liquid nitrogen. The harvested tissue is stored at
-80.degree. C. until RNA preparation. To make RNA, the three sample
timepoints were combined and ground. For subtraction, target cDNA
is made from the jasmonic acid treated tissue total RNA using the
SMART cDNA synthesis system from Clonetech (Clonetech Laboratories,
Palo Alto, Calif. U.S.A.). Driver first strand cDNA is covalently
linked to Dynabeads following a protocol similar to that described
in the Dynal literature (Dynabeads, Dynal Corporation, Lake
Success, N.Y. U.S.A.). The target cDNA is then heat denatured and
the second strand trapped using Dynabeads oligo-dT. The target
second strand cDNA is then hybridized to the driver cDNA in 400
.mu.l 2.times.SSPE for two rounds of hybridization at 65.degree. C.
and 20 hours. After each hybridization, the hybridization solution
is removed from the system and the hybridized target cDNA removed
from the driver by heat denaturation in water. After hybridization,
the remaining cDNA is trapped with Dynabeads oligo-dT. The trapped
cDNA is then amplified as in previous PCR based libraries and the
resulting cDNA ligated into the pSPORT vector (Invitrogen, Carlsbad
Calif. U.S.A.). For this library's construction, the eighth
fraction of the cDNA size fractionation step was used for
ligation.
[0478] The Soy62 (LIB3074) cDNA library is generated from soybean
cultivar Asgrow 3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.)
jasmonic acid treated seedling subtracted from control tissue.
Seeds are planted at a depth of approximately 2 cm into 2-3 inch
peat pots containing Metromix 350 medium and the plants are grown
in a greenhouse. The daytime temperature is approximately
29.4.degree. C. and the nighttime temperature 20.degree. C. Soil is
checked and watered daily to maintain even moisture conditions. At
9 days post planting, the plantlets are sprayed with either control
buffer of 0.1% Tween-20 or jasmonic acid (Sigma J-2500, Sigma, St.
Loius, Mo. U.S.A.) at 1 mg/ml in 0.1% Tween-20. Plants are sprayed
until runoff and the soil and the stem is socked with the spraying
solution. At 18 hours post application of jasmonic acid, the
soybean plantlets appear growth retarded. After 18 hours, 24 hours
and 48 hours post treatment, the cotyledons are removed and the
remaining leaf and stem tissue above the soil is harvested and
frozen in liquid nitrogen. The harvested tissue is stored at
-80.degree. C. until RNA preparation. To make RNA, the three sample
timepoints were combined and ground. For subtraction, target cDNA
is made from the jasmonic acid treated tissue total RNA using the
SMART cDNA synthesis system from Clonetech (Clonetech Laboratories,
Palo Alto, Calif. U.S.A.). Driver first strand cDNA is covalently
linked to Dynabeads following a protocol similar to that described
in the Dynal literature (Dynabeads, Dynal Corporation, Lake
Success, N.Y. U.S.A.). The target cDNA is then heat denatured and
the second strand trapped using Dynabeads oligo-dT. The target
second strand cDNA is then hybridized to the driver cDNA in 400
.mu.l 2.times.SSPE for two rounds of hybridization at 65.degree. C.
and 20 hours. After each hybridization, the hybridization solution
is removed from the system and the hybridized target cDNA removed
from the driver by heat denaturation in water. After hybridization,
the remaining cDNA is trapped with Dynabeads oligo-dT. The trapped
cDNA is then amplified as in previous PCR based libraries and the
resulting cDNA ligated into the pSPORT vector (Invitrogen, Carlsbad
Calif. U.S.A.). For this library's construction, the ninth fraction
of the cDNA size fractionation step was used for ligation.
[0479] The Soy65 (LIB3107) 07cDNA library is generated from soybean
cultivar Asgrow 3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.)
drought-stressed abscission zone tissue. Seeds are planted at a
depth of approximately 2 cm into 2-3 inch peat pots containing
Metromix 350 medium and the plants are grown in an environmental
chamber under 12 hr daytime/12 hr nighttime cycles. The daytime
temperature is approximately 29.degree. C. and the nighttime
temperature 24.degree. C. Soil is checked and watered daily to
maintain even moisture conditions. Plants are irrigated with
15-16-17 Peter's Mix. At the R3 stage of development, drought is
imposed by withholding water. At 3, 4, 5 and 6 days, tissue is
harvested and wilting is not obvious until the fourth day.
Abscission layers from reproductive organs are harvested by cutting
less than one millimeter proximal and distal to the layer and
immediately frozen in liquid nitrogen. The harvested tissue is
stored at -80.degree. C. until RNA preparation.
[0480] The Soy66 (LIB3109) cDNA library is generated from soybean
cultivar Asgrow 3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.)
non-drought stressed abscission zone tissue. Seeds are planted at a
depth of approximately 2 cm into 2-3 inch peat pots containing
Metromix 350 medium and the plants are grown in an environmental
chamber under 12 hr daytime/12 hr nighttime cycles. The daytime
temperature is approximately 29.degree. C. and the nighttime
temperature approximately 24.degree. C. Soil is checked and watered
daily to maintain even moisture conditions. Plants are irrigated
with 15-16-17 Peter's Mix. At 3, 4, 5 and 6 days, control
abscission layer tissue is harvested. Abscission layers from
reproductive organs are harvested by cutting less than one
millimeter proximal and distal to the layer and immediately frozen
in liquid nitrogen. The harvested tissue is stored at -80.degree.
C. until RNA preparation.
[0481] Soy67 (LIB3065) cDNA library is prepared from equal amounts
tissue harvested from SOYMON007, SOYMON015 and SOYMON020 prepared
tissue. Single stranded and double stranded DNA representing
approximately 1.times.10.sup.6 colony forming units are isolated
using standard protocols. RNA, complementary to the single stranded
DNA, is synthesized using the double stranded DNA as a template.
Biotinylated dATP is incorporated into the RNA during the synthesis
reaction. The single stranded DNA is mixed with the biotinylated
RNA in a 1:10 molar ratio) and allowed to hybridize. DNA-RNA
hybrids are captured on Dynabeads M280 streptavidin (Dynabeads,
Dynal Corporation, Lake Success, N.Y. U.S.A.). The dynabeads with
captured hybrids are collected with a magnet. Captured hybrids are
eluted with water.
[0482] Soy68 (LIB3052) cDNA library is prepared from equal amounts
tissue harvested from SOYMON007, SOYMON015 and SOYMON020 prepared
tissue. Single stranded and double stranded DNA representing
approximately 1.times.10.sup.6 colony forming units are isolated
using standard protocols. RNA, complementary to the single stranded
DNA, is synthesized using the double stranded DNA as a template.
Biotinylated dATP is incorporated into the RNA during the synthesis
reaction. The single stranded DNA is mixed with the biotinylated
RNA in a 1:10 molar ratio) and allowed to hybridize. DNA-RNA
hybrids are captured on Dynabeads M280 streptavidin (Dynabeads,
Dynal Corporation, Lake Success, N.Y. U.S.A.). The dynabeads with
captured hybrids are collected with a magnet. Captured hybrids are
eluted with water.
[0483] Soy69 (LIB3053) cDNA library is generated from soybean
cultivars Cristalina (USDA Soybean Germplasm Collection, Urbana,
Ill. U.S.A.) and FT108 (Monsoy, Brazil) (tropical germ plasma)
normalized leaf tissue. Leaves are harvested from plants grown in
an environmental chamber under 12 hr daytime/12 hr nighttime
cycles. The daytime temperature is approximately 29.degree. C. and
the nighttime temperature approximately 24.degree. C. Soil is
checked and watered daily to maintain even moisture conditions.
Approximately 30 g of leaves are harvested from the 4.sup.th node
of each of the Cristalina and FT108 cultivars and immediately
frozen in dry ice. The harvested tissue is then stored at
-80.degree. C. until RNA preparation. Single stranded and double
stranded DNA representing approximately 1.times.10.sup.6 colony
forming units are isolated using standard protocols. RNA,
complementary to the single stranded DNA, is synthesized using the
double stranded DNA as a template. Biotinylated dATP is
incorporated into the RNA during the synthesis reaction. The single
stranded DNA is mixed with the biotinylated RNA in a 1:10 molar
ratio) and allowed to hybridize. DNA-RNA hybrids are captured on
Dynabeads M280 streptavidin (Dynabeads, Dynal Corporation, Lake
Success, N.Y. U.S.A.). The dynabeads with captured hybrids are
collected with a magnet. The non-hybridized single stranded
molecules remaining after hybrid capture are converted to double
stranded form and represent the primary normalized library.
[0484] Soy70 (LIB3055) cDNA library is generated from soybean
cultivars Cristalina (USDA Soybean Germplasm Collection, Urbana,
Ill. U.S.A.) and FT108 (Monsoy, Brazil) (tropical germ plasma) leaf
tissue. Leaves are harvested from plants grown in an environmental
chamber under 12 hr daytime/12 hr nighttime cycles. The daytime
temperature is approximately 29.degree. C. and the nighttime
temperature approximately 24.degree. C. Soil is checked and watered
daily to maintain even moisture conditions. Approximately 30 g of
leaves are harvested from the 4.sup.th node of each of the
Cristalina and FT108 cultivars and immediately frozen in dry ice.
The harvested tissue is then stored at -80.degree. C. until RNA
preparation.
[0485] Soy71 (LIB3056) cDNA library is generated from soybean
cultivars Cristalina and FT108 (tropical germ plasma) root tissue.
Roots are harvested from plants grown in an environmental chamber
under 12 hr daytime/12 hr nighttime cycles. The daytime temperature
is approximately 29.degree. C. and the nighttime temperature
approximately 24.degree. C. Soil is checked and watered daily to
maintain even moisture conditions. Approximately 50 g and 56 g of
roots are harvested from each of the Cristalina and FT108 cultivars
and immediately frozen in dry ice. The harvested tissue is then
stored at -80.degree. C. until RNA preparation.
[0486] Soy72 (LIB3093) cDNA library is generated from soybean
cultivar Asgrow 3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.)
drought stressed leaf control tissue. Seeds are planted at a depth
of approximately 2 cm into 2-3 inch peat pots containing Metromix
350 medium and the plants are grown in an environmental chamber
under 12 hr daytime/12 hr nighttime cycles. The daytime temperature
is approximately 26.degree. C. and the nighttime temperature
21.degree. C. and 70% relative humidity. Soil is checked and
watered daily to maintain even moisture conditions. At the R3 stage
of the plant drought is induced by withholding water. After 3 and 6
days seeds and pods from both drought stressed and control (watered
regularly) plants are collected from the fifth and sixth node and
frozen in dry-ice. The harvested tissue is stored at -80.degree. C.
until RNA preparation. For subtraction, target cDNA is made from
the drought stressed tissue total RNA using the SMART cDNA
synthesis system from Clonetech (Clonetech Laboratories, Palo Alto,
Calif. U.S.A.). Driver first strand cDNA is covalently linked to
Dynabeads following a protocol similar to that described in the
Dynal literature (Dynabeads, Dynal Corporation, Lake Success, N.Y.
U.S.A.). The target cDNA is then heat denatured and the second
strand trapped using Dynabeads oligo-dT. The target second strand
cDNA is then hybridized to the driver cDNA in 400 .mu.l
2.times.SSPE for two rounds of hybridization at 65.degree. C. and
20 hours. After each hybridization, the hybridization solution is
removed from the system and the hybridized target cDNA removed from
the driver by heat denaturation in water. After hybridization, the
remaining cDNA is trapped with Dynabeads oligo-dT. The trapped cDNA
is then amplified as in previous PCR based libraries and the
resulting cDNA ligated into the pSPORT vector (Invitrogen, Carlsbad
Calif. U.S.A.).
[0487] Soy73 (LIB3093) cDNA library is generated from soybean
cultivar Asgrow 3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.)
drought stressed leaf subtracted from control tissue. Seeds are
planted at a depth of approximately 2 cm into 2-3 inch peat pots
containing Metromix 350 medium and the plants are grown in an
environmental chamber under 12 hr daytime/12 hr nighttime cycles.
The daytime temperature is approximately 26.degree. C. and the
nighttime temperature 21.degree. C. and 70% relative humidity. Soil
is checked and watered daily to maintain even moisture conditions.
At the R3 stage of the plant drought is induced by withholding
water. After 3 and 6 days seeds and pods from both drought stressed
and control (watered regularly) plants are collected from the fifth
and sixth node and frozen in dry-ice. The harvested tissue is
stored at -80.degree. C. until RNA preparation. For subtraction,
target cDNA is made from the drought stressed tissue total RNA
using the SMART cDNA synthesis system from Clonetech (Clonetech
Laboratories, Palo Alto, Calif. U.S.A.). Driver first strand cDNA
is covalently linked to Dynabeads following a protocol similar to
that described in the Dynal literature (Dynabeads, Dynal
Corporation, Lake Success, N.Y. U.S.A.). The target cDNA is then
heat denatured and the second strand trapped using Dynabeads
oligo-dT. The target second strand cDNA is then hybridized to the
driver cDNA in 400 .mu.l 2.times.SSPE for two rounds of
hybridization at 65.degree. C. and 20 hours. After each
hybridization, the hybridization solution is removed from the
system and the hybridized target cDNA removed from the driver by
heat denaturation in water. After hybridization, the remaining cDNA
is trapped with Dynabeads oligo-dT. The trapped cDNA is then
amplified as in previous PCR based libraries and the resulting cDNA
ligated into the pSPORT vector (Invitrogen, Carlsbad Calif.
U.S.A.).
[0488] The Soy76 (Lib3106) cDNA library is generated from soybean
cultivar Asgrow 3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.)
jasmonic acid and arachidonic treated seedling subtracted from
control tissue. Seeds are planted at a depth of approximately 2 cm
into 2-3 inch peat pots containing Metromix 350 medium and the
plants are grown in a greenhouse. The daytime temperature is
approximately 29.4.degree. C. and the nighttime temperature
20.degree. C. Soil is checked and watered daily to maintain even
moisture conditions. At 9 days post planting, the plantlets are
sprayed with either control buffer of 0.1% Tween-20 or jasmonic
acid (Sigma J-2500, Sigma, St. Loius, Mo. U.S.A.) at 1 mg/ml in
0.1% Tween-20. Plants are sprayed until runoff and the soil and the
stem is socked with the spraying solution. At 18 hours post
application of jasmonic acid, the soybean plantlets appear growth
retarded. Arachidonic treated seedlings are sprayed with 1 m/ml
arachidonic acid in 0.1% Tween-20. After 18 hours, 24 hours and 48
hours post treatment, the cotyledons are removed and the remaining
leaf and stem tissue above the soil is harvested and frozen in
liquid nitrogen. The harvested tissue is stored at -80.degree. C.
until RNA preparation. To make RNA, the three sample timepoints
were combined and ground. The RNA from the arachidonic treated
seedlings is isolated separately. For subtraction, target cDNA is
made from the jasmonic acid treated tissue total RNA using the
SMART cDNA synthesis system from Clonetech (Clonetech Laboratories,
Palo Alto, Calif. U.S.A.). Driver first strand cDNA is covalently
linked to Dynabeads following a protocol similar to that described
in the Dynal literature (Dynabeads, Dynal Corporation, Lake
Success, N.Y. U.S.A.). The target cDNA is then heat denatured and
the second strand trapped using Dynabeads oligo-dT. The target
second strand cDNA is then hybridized to the driver cDNA in 400
.mu.l 2.times.SSPE for two rounds of hybridization at 65.degree. C.
and 20 hours. After each hybridization, the hybridization solution
is removed from the system and the hybridized target cDNA removed
from the driver by heat denaturation in water. After hybridization,
the remaining cDNA is trapped with Dynabeads oligo-dT. The trapped
cDNA is then amplified as in previous PCR based libraries and the
resulting cDNA ligated into the pSPORT vector (Invitrogen, Carlsbad
Calif. U.S.A.). Fraction 10 of the size fractionated cDNA is
ligated into the pSPORT vector (Invitrogen, Carlsbad Calif. U.S.A.)
in order to capture some of the smaller transcripts characteristic
of antifungal proteins.
[0489] Soy77 (LIB3108) cDNA library is generated from soybean
cultivar Asgrow 3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.)
jasmonic acid control tissue. Seeds are planted at a depth of
approximately 2 cm into 2-3 inch peat pots containing Metromix 350
medium and the plants are grown in a greenhouse. The daytime
temperature is approximately 29.4.degree. C. and the nighttime
temperature 20.degree. C. Soil is checked and watered daily to
maintain even moisture conditions. At 9 days post planting, the
plantlets are sprayed with either control buffer of 0.1% Tween-20
or jasmonic acid (Sigma J-2500, Sigma, St. Loius, Mo. U.S.A.) at 1
mg/ml in 0.1% Tween-20. Plants are sprayed until runoff and the
soil and the stem is socked with the spraying solution. At 18 hours
post application of jasmonic acid, the soybean plantlets appear
growth retarded. Arachidonic treated seedlings are sprayed with 1
m/ml arachidonic acid in 0.1% Tween-20. After 18 hours, 24 hours
and 48 hours post treatment, the cotyledons are removed and the
remaining leaf and stem tissue above the soil is harvested and
frozen in liquid nitrogen. The harvested tissue is stored at
-80.degree. C. until RNA preparation. To make RNA, the three sample
timepoints were combined and ground. The RNA from the arachidonic
treated seedlings is isolated separately. For subtraction, target
cDNA is made from the jasmonic acid treated tissue total RNA using
the SMART cDNA synthesis system from Clonetech (Clonetech
Laboratories, Palo Alto, Calif. U.S.A.). Driver first strand cDNA
is covalently linked to Dynabeads following a protocol similar to
that described in the Dynal literature (Dynabeads, Dynal
Corporation, Lake Success, N.Y. U.S.A.). The target cDNA is then
heat denatured and the second strand trapped using Dynabeads
oligo-dT. The target second strand cDNA is then hybridized to the
driver cDNA in 400 .mu.l 2.times.SSPE for two rounds of
hybridization at 65.degree. C. and 20 hours. After each
hybridization, the hybridization solution is removed from the
system and the hybridized target cDNA removed from the driver by
heat denaturation in water. After hybridization, the remaining cDNA
is trapped with Dynabeads oligo-dT. The trapped cDNA is then
amplified as in previous PCR based libraries and the resulting cDNA
ligated into the pSPORT vector (Invitrogen, Carlsbad Calif.
U.S.A.). Fraction 10 of the size fractionated cDNA is ligated into
the pSPORT vector in order to capture some of the smaller
transcripts characteristic of antifungal proteins.
[0490] The stored RNA is purified using Trizol reagent from Life
Technologies (Gibco BRL, Life Technologies, Gaithersburg, Md.
U.S.A.), essentially as recommended by the manufacturer. Poly A+
RNA (mRNA) is purified using magnetic oligo dT beads essentially as
recommended by the manufacturer (Dynabeads, Dynal Corporation, Lake
Success, N.Y. U.S.A.).
[0491] Construction of plant cDNA libraries is well-known in the
art and a number of cloning strategies exist. A number of cDNA
library construction kits are commercially available. The
Superscript.TM. Plasmid System for cDNA synthesis and Plasmid
Cloning (Gibco BRL, Life Technologies, Gaithersburg, Md. U.S.A.) is
used, following the conditions suggested by the manufacturer.
[0492] Normalized libraries are made using essentially the Soares
procedure (Soares et al., Proc. Natl. Acad. Sci. (U.S.A.)
91:9228-9232 (1994), the entirety of which is herein incorporated
by reference). This approach is designed to reduce the initial
10.000-fold variation in individual cDNA frequencies to achieve
abundances within one order of magnitude while maintaining the
overall sequence complexity of the library. In the normalization
process, the prevalence of high-abundance cDNA clones decreases
dramatically, clones with mid-level abundance are relatively
unaffected and clones for rare transcripts are effectively
increased in abundance.
Example 2
[0493] The cDNA libraries are plated on LB agar containing the
appropriate antibiotics for selection and incubated at 37.degree.
for a sufficient time to allow the growth of individual colonies.
Single colonies are individually placed in each well of a 96-well
microtiter plates containing LB liquid including the selective
antibiotics. The plates are incubated overnight at approximately
37.degree. C. with gentle shaking to promote growth of the
cultures. The plasmid DNA is isolated from each clone using Qiaprep
plasmid isolation kits, using the conditions recommended by the
manufacturer (Qiagen Inc., Santa Clara, Calif. U.S.A.).
[0494] Template plasmid DNA clones are used for subsequent
sequencing. For sequencing, the ABI PRISM dRhodamine Terminator
Cycle Sequencing Ready Reaction Kit with AmpliTaq.RTM. DNA
Polymerase, FS, is used (PE Applied Biosystems, Foster City, Calif.
U.S.A.).
Example 3
[0495] Nucleic acid sequences that encode for the following
proteins: methionine adenosyltransferase, S-adenosylmethionine
decarboxylase, aspartate kinase, aspartate-semialdehyde
dehydrogenase, O-succinylhomoserine (thiol)-lyase, cystathionine
.beta.-lyase, 5-methyltetrahydropteroyltriglutamate,
adenosylhomocysteinase, cystathionine .beta.-synthase,
cystathionine .gamma.-lyase and O-acetylhomoserine (thiol)-lyase
are identified from the Monsanto EST PhytoSeq database using
TBLASTN (default values)(TBLASTN compares a protein query against
the six reading frames of a nucleic acid sequence). Matches found
with BLAST P values equal or less than 0.001 (probability) or BLAST
Score of equal or greater than 90 are classified as hits. If the
program used to determine the hit is HMMSW then the score refers to
HMMSW score.
[0496] In addition, the GenBank database is searched with BLASTN
and BLASTX (default values) using ESTs as queries. EST that pass
the hit probability threshold of 10 e.sup.-8 for the following
enzymes are combined with the hits generated by using TBLASTN
(described above) and classified by enzyme (see Table A below).
[0497] A cluster refers to a set of overlapping clones in the
PhytoSeq database. Such an overlapping relationship among clones is
designated as a "cluster" when BLAST scores from pairwise sequence
comparisons of the member clones meets a predetermined minimum
value or product score of 50 or more (Product Score=(BLAST
SCORE.times.Percentage Identity)/(5.times. minimum [length (Seq1),
length (Seq2)]))
[0498] Since clusters are formed on the basis of single-linkage
relationships, it is possible for two non-overlapping clones to be
members of the same cluster if, for instance, they both overlap a
third clone with at least the predetermined minimum BLAST score
(stringency). A cluster ID is arbitrarily assigned to all of those
clones which belong to the same cluster at a given stringency and a
particular clone will belong to only one cluster at a given
stringency. If a cluster contains only a single clone (a
"singleton"), then the cluster ID number will be negative, with an
absolute value equal to the clone ID number of its single member.
Clones grouped in a cluster in most cases represent a contiguous
sequence.
TABLE-US-00002 TABLE A* Seq No. Cluster ID Clone ID Library NCBI gi
Method Score P-value % Ident METHIONINE ADENOSYLTRANSFERASE (EC
2.5.1.6) 1 -700019427 700019427H1 SATMON001 g17262 BLASTX 120 1e-11
92 2 -700074004 700074004H1 SATMON007 g1778820 BLASTN 830 1e-60 80
3 -700149718 700149718H1 SATMON007 g1778820 BLASTN 263 1e-13 80 4
-700220251 700220251H1 SATMON011 g1127582 BLASTN 243 1e-9 90 5
-700458196 700458196H1 SATMON029 g1778820 BLASTN 202 1e-22 88 6
-701172459 701172459H1 SATMONN05 g882334 BLASTN 801 1e-57 89 7
-L1436317 LIB143-062-Q1-E1-B6 LIB143 g1778820 BLASTN 341 1e-19 77 8
-L1893126 LIB189-021-Q1-E1-F4 LIB189 g1778820 BLASTN 629 1e-41 88 9
-L1894542 LIB189-033-Q1-E1-H12 LIB189 g1778820 BLASTN 564 1e-60 76
10 -L30622839 LIB3062-028-Q1-K1-C2 LIB3062 g1778820 BLASTN 625
1e-65 74 11 -L30671966 LIB3067-036-Q1-K1-D8 LIB3067 g1655576 BLASTX
118 1e-25 85 12 -L30682166 LIB3068-004-Q1-K1-D5 LIB3068 g497900
BLASTX 111 1e-29 47 13 1 LIB143-036-Q1-E1-G7 LIB143 g450548 BLASTN
1766 1e-138 88 14 1 LIB3060-013-Q1-K1-F7 LIB3060 g1778820 BLASTN
1702 1e-133 90 15 1 LIB143-013-Q1-E1-A6 LIB143 g1778820 BLASTN 1712
1e-133 88 16 1 LIB148-007-Q1-E1-B8 LIB148 g450548 BLASTN 1693
1e-132 88 17 1 LIB3079-003-Q1-K1-G1 LIB3079 g1778820 BLASTN 1236
1e-130 88 18 1 LIB3066-011-Q1-K1-E4 LIB3066 g1778820 BLASTN 1661
1e-129 86 19 1 LIB3068-008-Q1-K1-D2 LIB3068 g450548 BLASTN 1192
1e-128 88 20 1 LIB143-061-Q1-E1-E7 LIB143 g1778820 BLASTN 1638
1e-127 88 21 1 LIB189-006-Q1-E1-B2 LIB189 g1778820 BLASTN 1627
1e-126 86 22 1 LIB143-031-Q1-E1-C7 LIB143 g450548 BLASTN 1595
1e-124 86 23 1 LIB3068-011-Q1-K1-B12 LIB3068 g450548 BLASTN 1552
1e-123 87 24 1 LIB143-013-Q1-E1-G11 LIB143 g450548 BLASTN 1585
1e-123 88 25 1 LIB3062-057-Q1-K1-H9 LIB3062 g450548 BLASTN 1577
1e-122 89 26 1 LIB143-008-Q1-E1-G9 LIB143 g450548 BLASTN 1556
1e-120 89 27 1 LIB3068-058-Q1-K1-B7 LIB3068 g450548 BLASTN 1470
1e-118 86 28 1 LIB3067-048-Q1-K1-G5 LIB3067 g1778820 BLASTN 1518
1e-117 89 29 1 LIB143-049-Q1-E1-A12 LIB143 g1778820 BLASTN 1480
1e-114 91 30 1 LIB3062-010-Q1-K1-C7 LIB3062 g1778820 BLASTN 1145
1e-113 87 31 1 LIB3068-055-Q1-K1-E10 LIB3068 g450548 BLASTN 1451
1e-112 88 32 1 LIB189-025-Q1-E1-H1 LIB189 g1778820 BLASTN 1460
1e-112 90 33 1 LIB143-066-Q1-E1-F4 LIB143 g1778820 BLASTN 1309
1e-111 86 34 1 LIB143-003-Q1-E1-D10 LIB143 g1778820 BLASTN 1440
1e-111 87 35 1 LIB3062-033-Q1-K1-G1 LIB3062 g1778820 BLASTN 1449
1e-111 86 36 1 LIB3066-029-Q1-K1-H11 LIB3066 g1778820 BLASTN 1427
1e-110 88 37 1 LIB3068-016-Q1-K1-D9 LIB3068 g450548 BLASTN 1213
1e-109 87 38 1 LIB3062-027-Q1-K1-B11 LIB3062 g1778820 BLASTN 1187
1e-108 87 39 1 LIB3068-048-Q1-K1-G4 LIB3068 g1778820 BLASTN 1410
1e-108 82 40 1 LIB148-020-Q1-E1-B6 LIB148 g1778820 BLASTN 1088
1e-107 84 41 1 LIB3066-046-Q1-K1-F2 LIB3066 g1778820 BLASTN 1400
1e-107 90 42 1 700084130H1 SATMON011 g1778820 BLASTN 960 1e-106 92
43 1 700092863H1 SATMON008 g1778820 BLASTN 1388 1e-106 92 44 1
LIB3068-043-Q1-K1-A12 LIB3068 g450548 BLASTN 827 1e-105 81 45 1
700092659H1 SATMON008 g1778820 BLASTN 1359 1e-104 92 46 1
LIB143-038-Q1-E1-H11 LIB143 g450548 BLASTN 1345 1e-103 84 47 1
LIB3062-028-Q1-K1-F12 LIB3062 g1778820 BLASTN 1082 1e-102 82 48 1
700103135H1 SATMON010 g1778820 BLASTN 1306 1e-100 93 49 1
LIB3078-050-Q1-K1-D9 LIB3078 g1778820 BLASTN 1309 1e-100 86 50 1
700084823H1 SATMON011 g1778820 BLASTN 1311 1e-100 90 51 1
700201311H1 SATMON003 g1778820 BLASTN 960 1e-97 89 52 1 700265914H1
SATMON017 g1778820 BLASTN 1275 1e-97 91 53 1 700205948H1 SATMON003
g1778820 BLASTN 983 1e-96 93 54 1 LIB189-003-Q1-E1-A6 LIB189
g450548 BLASTN 1262 1e-96 86 55 1 LIB143-017-Q1-E1-B9 LIB143
g1778820 BLASTN 779 1e-95 86 56 1 700097434H1 SATMON009 g450548
BLASTN 1247 1e-95 90 57 1 700089089H1 SATMON011 g450548 BLASTN 1251
1e-95 89 58 1 700071949H1 SATMON007 g1778820 BLASTN 1256 1e-95 87
59 1 700047892H1 SATMON003 g1778820 BLASTN 1236 1e-94 91 60 1
700077246H1 SATMON007 g450548 BLASTN 1237 1e-94 89 61 1 700085681H1
SATMON011 g1778820 BLASTN 1237 1e-94 89 62 1 LIB3067-010-Q1-K1-A7
LIB3067 g1778820 BLASTN 1244 1e-94 83 63 1 700619961H1 SATMON034
g450548 BLASTN 962 1e-93 89 64 1 700087395H1 SATMON011 g450548
BLASTN 1040 1e-93 89 65 1 700240431H1 SATMON010 g1778820 BLASTN
1156 1e-93 92 66 1 700053822H1 SATMON011 g1778820 BLASTN 1224 1e-93
91 67 1 700103856H1 SATMON010 g450548 BLASTN 1224 1e-93 88 68 1
700083703H1 SATMON011 g450548 BLASTN 1229 1e-93 89 69 1 700087360H1
SATMON011 g450548 BLASTN 1234 1e-93 90 70 1 700104713H1 SATMON010
g450548 BLASTN 1188 1e-92 87 71 1 700265996H1 SATMON017 g450548
BLASTN 1214 1e-92 88 72 1 700264870H1 SATMON017 g450548 BLASTN 945
1e-91 89 73 1 700219348H1 SATMON011 g1778820 BLASTN 1203 1e-91 91
74 1 700095527H1 SATMON008 g450548 BLASTN 1153 1e-90 88 75 1
LIB3068-023-Q1-K1-G5 LIB3068 g1778820 BLASTN 1187 1e-90 89 76 1
700102547H1 SATMON010 g1778820 BLASTN 1194 1e-90 88 77 1
700332179H1 SATMON019 g450548 BLASTN 1054 1e-89 88 78 1 700405470H1
SATMON029 g450548 BLASTN 1071 1e-89 90 79 1 LIB3069-043-Q1-K1-G2
LIB3069 g450548 BLASTN 1175 1e-89 84 80 1 700451332H1 SATMON028
g450548 BLASTN 1176 1e-89 90 81 1 700028108H1 SATMON003 g450548
BLASTN 1180 1e-89 89 82 1 700076154H1 SATMON007 g1778820 BLASTN
1180 1e-89 89 83 1 700089770H1 SATMON011 g450548 BLASTN 1185 1e-89
89 84 1 700051882H1 SATMON003 g1778820 BLASTN 844 1e-88 91 85 1
700242942H1 SATMON010 g1778820 BLASTN 1163 1e-88 91 86 1
700094996H1 SATMON008 g1778820 BLASTN 1166 1e-88 88 87 1
700476239H1 SATMON025 g450548 BLASTN 1166 1e-88 89 88 1 700219530H1
SATMON011 g1778820 BLASTN 1005 1e-87 91 89 1 700049726H1 SATMON003
g1778820 BLASTN 1041 1e-87 89 90 1 700096904H1 SATMON008 g1778820
BLASTN 1157 1e-87 86 91 1 700344026H1 SATMON021 g450548 BLASTN 1158
1e-87 88 92 1 700105590H1 SATMON010 g1778820 BLASTN 1139 1e-86 89
93 1 700465283H1 SATMON025 g450548 BLASTN 1145 1e-86 89 94 1
700104341H1 SATMON010 g450548 BLASTN 1146 1e-86 87 95 1
LIB3062-021-Q1-K1-F12 LIB3062 g1778820 BLASTN 923 1e-85 78 96 1
700105246H1 SATMON010 g450548 BLASTN 930 1e-85 89 97 1 700072146H1
SATMON007 g1778820 BLASTN 1131 1e-85 91 98 1 700028553H1 SATMON003
g1778820 BLASTN 1132 1e-85 88 99 1 700451419H1 SATMON028 g450548
BLASTN 1133 1e-85 89 100 1 700103241H1 SATMON010 g450548 BLASTN
1134 1e-85 90 101 1 700612549H1 SATMON033 g450548 BLASTN 1134 1e-85
89 102 1 700050434H1 SATMON003 g450548 BLASTN 723 1e-84 89 103 1
700094764H1 SATMON008 g1778820 BLASTN 977 1e-84 86 104 1
700163030H1 SATMON013 g1778820 BLASTN 1011 1e-84 93 105 1
700466542H1 SATMON025 g450548 BLASTN 1085 1e-84 89 106 1
700047380H1 SATMON003 g450548 BLASTN 1096 1e-84 89 107 1
700456429H1 SATMON029 g450548 BLASTN 1118 1e-84 88 108 1
700096049H1 SATMON008 g1778820 BLASTN 1122 1e-84 86 109 1
700075906H1 SATMON007 g1778820 BLASTN 838 1e-83 86 110 1
LIB3067-048-Q1-K1-G3 LIB3067 g1778820 BLASTN 1045 1e-83 86 111 1
700214043H1 SATMON016 g450548 BLASTN 1103 1e-83 88 112 1
700096909H1 SATMON008 g1778820 BLASTN 1103 1e-83 90 113 1
701184635H1 SATMONN06 g1778820 BLASTN 1104 1e-83 87 114 1
700158969H1 SATMON012 g1778820 BLASTN 1107 1e-83 93 115 1
700475902H1 SATMON025 g450548 BLASTN 1112 1e-83 89 116 1
700207076H1 SATMON003 g1778820 BLASTN 1113 1e-83 90 117 1
700161901H1 SATMON012 g1778820 BLASTN 1091 1e-82 91 118 1
700452183H1 SATMON028 g450548 BLASTN 1095 1e-82 88 119 1
700452326H1 SATMON028 g450548 BLASTN 1098 1e-82 88 120 1
700106918H1 SATMON010 g450548 BLASTN 1101 1e-82 88 121 1
700220146H1 SATMON011 g450548 BLASTN 1083 1e-81 89 122 1
700221060H1 SATMON011 g450548 BLASTN 1084 1e-81 88 123 1
700243344H1 SATMON010 g1778820 BLASTN 1084 1e-81 92 124 1
700475790H1 SATMON025 g450548 BLASTN 1084 1e-81 89 125 1
700452691H1 SATMON028 g1778820 BLASTN 1086 1e-81 87 126 1
700243733H1 SATMON010 g450548 BLASTN 1086 1e-81 90 127 1
LIB3059-018-Q1-K1-D10 LIB3059 g1778820 BLASTN 830 1e-80 91 128 1
700165958H1 SATMON013 g450548 BLASTN 1070 1e-80 89 129 1
700082659H1 SATMON011 g1778820 BLASTN 1071 1e-80 88 130 1
700172557H1 SATMON013 g1778820 BLASTN 1074 1e-80 92 131 1
700221526H1 SATMON011 g450548 BLASTN 1075 1e-80 88 132 1
700456696H1 SATMON029 g450548 BLASTN 1077 1e-80 89 133 1
700430776H1 SATMONN01 g1778820 BLASTN 574 1e-79 90 134 1
LIB3062-025-Q1-K1-B11 LIB3062 g450548 BLASTN 585 1e-79 85 135 1
700077126H1 SATMON007 g1778820 BLASTN 743 1e-79 88 136 1
700160310H1 SATMON012 g1778820 BLASTN 752 1e-79 91 137 1
700030466H1 SATMON003 g1778820 BLASTN 757 1e-79 83 138 1
700210204H1 SATMON016 g1778820 BLASTN 844 1e-79 91 139 1
700094716H1 SATMON008 g1778820 BLASTN 857 1e-79 86 140 1
700157051H1 SATMON012 g1778820 BLASTN 937 1e-79 89 141 1
700241415H1 SATMON010 g450548 BLASTN 1055 1e-79 90 142 1
700160127H1 SATMON012 g450548 BLASTN 1056 1e-79 88 143 1
700205485H1 SATMON003 g1778820 BLASTN 1059 1e-79 90 144 1
700475930H1 SATMON025 g450548 BLASTN 1060 1e-79 89 145 1
700161902H1 SATMON012 g1778820 BLASTN 1060 1e-79 91 146 1
700213573H1 SATMON016 g450548 BLASTN 1061 1e-79 89 147 1
700207677H1 SATMON016 g450548 BLASTN 1061 1e-79 88 148 1
700581117H1 SATMON031 g450548 BLASTN 1065 1e-79 88 149 1
700451418H1 SATMON028 g450548 BLASTN 546 1e-78 89 150 1 700212606H1
SATMON016 g1778820 BLASTN 1044 1e-78 85 151 1 700222605H1 SATMON011
g1778820 BLASTN 1047 1e-78 86 152 1 700050542H1 SATMON003 g1778820
BLASTN 1053 1e-78 86 153 1 700450709H1 SATMON028 g450548 BLASTN 834
1e-77 88 154 1 700213368H1 SATMON016 g450548 BLASTN 861 1e-77 89
155 1 700095926H1 SATMON008 g1778820 BLASTN 1030 1e-77 86 156 1
700261686H1 SATMON017 g1778820 BLASTN 1034 1e-77 89 157 1
700235993H1 SATMON010 g450548 BLASTN 1037 1e-77 88 158 1
700466152H1 SATMON025 g450548 BLASTN 1038 1e-77 89 159 1
700266410H1 SATMON017 g1778820 BLASTN 1041 1e-77 85 160 1
700216991H1 SATMON016 g1778820 BLASTN 491 1e-76 92 161 1
700464957H1 SATMON025 g450548 BLASTN 607 1e-76 88 162 1 700160463H1
SATMON012 g1778820 BLASTN 1018 1e-76 91 163 1 700158224H1 SATMON012
g1778820 BLASTN 1018 1e-76 92 164 1 700469971H1 SATMON025 g1778820
BLASTN 1020 1e-76 86 165 1 700222731H1 SATMON011 g450548 BLASTN
1022 1e-76 88 166 1 700087434H1 SATMON011 g1778820 BLASTN 1023
1e-76 86 167 1 700094482H1 SATMON008 g1778820 BLASTN 1023 1e-76 86
168 1 700235976H1 SATMON010 g450548 BLASTN 1028 1e-76 89 169 1
700088266H1 SATMON011 g1778820 BLASTN 983 1e-75 92 170 1
700075882H1 SATMON007 g1778820 BLASTN 1009 1e-75 83 171 1
700243324H1 SATMON010 g1778820 BLASTN 1012 1e-75 86 172 1
700159625H2 SATMON012 g450548 BLASTN 1015 1e-75 90 173 1
700204422H1 SATMON003 g1778820 BLASTN 1015 1e-75 88 174 1
700048228H1 SATMON003 g1778820 BLASTN 741 1e-74 85 175 1
700072474H1 SATMON007 g1778820 BLASTN 799 1e-74 85 176 1
700477790H1 SATMON025 g450548 BLASTN 960 1e-74 89 177 1
LIB3059-022-Q1-K1-A9 LIB3059 g1778820 BLASTN 995 1e-74 86 178 1
700241633H1 SATMON010 g450548 BLASTN 997 1e-74 89 179 1 700093978H1
SATMON008 g1778820 BLASTN 999 1e-74 89 180 1 700238569H1 SATMON010
g450548 BLASTN 844 1e-73 88 181 1 700455160H1 SATMON029 g450548
BLASTN 849 1e-73 86 182 1 700262745H1 SATMON017 g1778820 BLASTN 984
1e-73 85 183 1 700405029H1 SATMON027 g1778820 BLASTN 990 1e-73 90
184 1 700195230H1 SATMON014 g450548 BLASTN 991 1e-73 88 185 1
700264753H1 SATMON017 g1778820 BLASTN 993 1e-73 90 186 1
700050142H1 SATMON003 g1778820 BLASTN 433 1e-72 86 187 1
700452677H1 SATMON028 g450548 BLASTN 919 1e-72 87 188 1 700168870H1
SATMON013 g1778820 BLASTN 971 1e-72 90 189 1 700023193H1 SATMON003
g450548 BLASTN 972 1e-72 88 190 1 700086582H1 SATMON011 g1778820
BLASTN 980 1e-72 84 191 1 700457710H1 SATMON029 g450548 BLASTN 982
1e-72 87 192 1 700477601H1 SATMON025 g450548 BLASTN 635 1e-71 88
193 1 700377730H1 SATMON019 g1778820 BLASTN 691 1e-71 87 194 1
700169782H1 SATMON013 g1778820 BLASTN 960 1e-71 91 195 1
700461113H1 SATMON033 g1778820 BLASTN 962 1e-71 85 196 1
700258669H1 SATMON017 g1778820 BLASTN 963 1e-71 90 197 1
700454253H1 SATMON029 g1778820 BLASTN 964 1e-71 85 198 1
700092376H1 SATMON008 g1778820 BLASTN 780 1e-70 83 199 1
700242885H1 SATMON010 g960356 BLASTN 946 1e-70 86 200 1 700094333H1
SATMON008 g1778820 BLASTN 947 1e-70 86 201 1 700575124H1 SATMON030
g450548 BLASTN 948 1e-70 82 202 1 700072544H1 SATMON007 g1778820
BLASTN 949 1e-70 86 203 1 LIB143-046-Q1-E1-B5 LIB143 g450548 BLASTN
951 1e-70 87 204 1 700096534H1 SATMON008 g1778820 BLASTN 956 1e-70
85 205 1 700262906H1 SATMON017 g1778820 BLASTN 937 1e-69 87 206 1
700094478H1 SATMON008 g1778820 BLASTN 939 1e-69 90 207 1
700172985H1 SATMON013 g450548 BLASTN 939 1e-69 88 208 1 700097938H1
SATMON009 g1778820 BLASTN 941 1e-69 85 209 1 LIB3069-006-Q1-K1-D5
LIB3069 g450548 BLASTN 943 1e-69 88 210 1 700093125H1 SATMON008
g1778820 BLASTN 945 1e-69 88 211 1 700424156H1 SATMONN01 g450548
BLASTN 752 1e-68 85 212 1 700405486H1 SATMON029 g450548 BLASTN 864
1e-68 91 213 1 LIB189-026-Q1-E1-H12 LIB189 g1778820 BLASTN 879
1e-68 87 214 1 700106172H1 SATMON010 g1778820 BLASTN 925 1e-68 85
215 1 700096890H1 SATMON008 g1778820 BLASTN 926 1e-68 90 216 1
700154404H1 SATMON007 g450548 BLASTN 928 1e-68 89 217 1 700468337H1
SATMON025 g450548 BLASTN 445 1e-67 88 218 1 700158827H1 SATMON012
g1778820 BLASTN 522 1e-67 87 219 1 700241744H1 SATMON010 g1778820
BLASTN 575 1e-67 85 220 1 700624633H1 SATMON034 g960356 BLASTN 644
1e-67 85 221 1 700154564H1 SATMON007 g1778820 BLASTN 735 1e-67 85
222 1 700172424H1 SATMON013 g450548 BLASTN 912 1e-67 90 223 1
700096287H1 SATMON008 g1778820 BLASTN 914 1e-67 86 224 1
700207638H1 SATMON016 g1778820 BLASTN 914 1e-67 86 225 1
700093735H1 SATMON008 g960356 BLASTN 920 1e-67 89 226 1 700159494H1
SATMON012 g1778820 BLASTN 899 1e-66 89 227 1 700236013H1 SATMON010
g1778820 BLASTN 900 1e-66 83 228 1 700220267H1 SATMON011 g1778820
BLASTN 439 1e-65 84 229 1 700477578H1 SATMON025 g450548 BLASTN 576
1e-65 90 230 1 700047743H1 SATMON003 g1778820 BLASTN 601 1e-65 82
231 1 700343041H1 SATMON021 g1778820 BLASTN 633 1e-65 86 232 1
700159886H1 SATMON012 g450548 BLASTN 803 1e-65 85 233 1 700570242H1
SATMON030 g450548 BLASTN 834 1e-65 85 234 1 700021930H1 SATMON001
g1778820 BLASTN 888 1e-65 87 235 1 700454959H1 SATMON029 g450548
BLASTN 890 1e-65 89 236 1 700171336H1 SATMON013 g1778820 BLASTN 890
1e-65 89 237 1 700105361H1 SATMON010 g1778820 BLASTN 806 1e-64 87
238 1 LIB3068-031-Q1-K1-B1 LIB3068 g450548 BLASTN 853 1e-64 89 239
1 700236324H1 SATMON010 g450548 BLASTN 875 1e-64 89 240 1
700150620H1 SATMON007 g450548 BLASTN 880 1e-64 87 241 1 700220648H1
SATMON011 g450548 BLASTN 881 1e-64 90 242 1 700150733H1 SATMON007
g450548 BLASTN 881 1e-64 85 243 1 700157367H1 SATMON012 g1778820
BLASTN 882 1e-64 85
244 1 700259676H1 SATMON017 g1778820 BLASTN 885 1e-64 88 245 1
700616490H1 SATMON033 g450548 BLASTN 886 1e-64 82 246 1 700102511H1
SATMON010 g450548 BLASTN 695 1e-63 89 247 1 700202805H1 SATMON003
g1778820 BLASTN 817 1e-63 92 248 1 700105685H1 SATMON010 g1778820
BLASTN 866 1e-63 84 249 1 700106113H1 SATMON010 g450548 BLASTN 873
1e-63 91 250 1 700444778H1 SATMON027 g1778820 BLASTN 343 1e-62 84
251 1 700103584H1 SATMON010 g450548 BLASTN 521 1e-62 86 252 1
LIB189-008-Q1-E1-D9 LIB189 g1778820 BLASTN 850 1e-62 91 253 1
700155684H1 SATMON007 g1778820 BLASTN 852 1e-62 85 254 1
700261639H1 SATMON017 g1778820 BLASTN 853 1e-62 89 255 1
700158367H1 SATMON012 g450548 BLASTN 856 1e-62 81 256 1 700153242H1
SATMON007 g1778820 BLASTN 859 1e-62 90 257 1 700210738H1 SATMON016
g1778820 BLASTN 859 1e-62 90 258 1 700203008H1 SATMON003 g450548
BLASTN 698 1e-61 86 259 1 700206081H1 SATMON003 g1778820 BLASTN 840
1e-61 92 260 1 700028643H1 SATMON003 g1778820 BLASTN 846 1e-61 85
261 1 700223914H1 SATMON011 g1778820 BLASTN 849 1e-61 84 262 1
700571455H1 SATMON030 g1778820 BLASTN 378 1e-60 88 263 1
700075374H1 SATMON007 g1778820 BLASTN 830 1e-60 85 264 1
700150452H1 SATMON007 g450548 BLASTN 831 1e-60 89 265 1 700261318H1
SATMON017 g1778820 BLASTN 831 1e-60 83 266 1 700616390H1 SATMON033
g450548 BLASTN 835 1e-60 92 267 1 700208718H1 SATMON016 g450548
BLASTN 561 1e-59 89 268 1 700448948H1 SATMON028 g1778820 BLASTN 653
1e-59 83 269 1 LIB3060-035-Q1-K1-H3 LIB3060 g450548 BLASTN 734
1e-59 85 270 1 700049753H1 SATMON003 g450548 BLASTN 769 1e-59 90
271 1 700154489H1 SATMON007 g1778820 BLASTN 814 1e-59 83 272 1
700170783H1 SATMON013 g1778820 BLASTN 816 1e-59 87 273 1
700237571H1 SATMON010 g450548 BLASTN 817 1e-59 89 274 1 700154872H1
SATMON007 g1778820 BLASTN 822 1e-59 85 275 1 700025620H1 SATMON004
g1778820 BLASTN 686 1e-58 84 276 1 700158255H1 SATMON012 g450548
BLASTN 803 1e-58 86 277 1 700159282H1 SATMON012 g450548 BLASTN 805
1e-58 83 278 1 700222171H1 SATMON011 g1778820 BLASTN 807 1e-58 89
279 1 700205003H1 SATMON003 g1778820 BLASTN 441 1e-57 86 280 1
700049209H1 SATMON003 g1778820 BLASTN 443 1e-57 90 281 1
700016430H1 SATMON001 g1778820 BLASTN 492 1e-57 86 282 1
700212936H1 SATMON016 g450548 BLASTN 564 1e-57 86 283 1 700156939H1
SATMON012 g1778820 BLASTN 801 1e-57 84 284 1 700222183H1 SATMON011
g1778820 BLASTN 561 1e-56 83 285 1 700203453H1 SATMON003 g1778820
BLASTN 784 1e-56 90 286 1 700167708H1 SATMON013 g1778820 BLASTN 560
1e-55 83 287 1 700214356H1 SATMON016 g1778820 BLASTN 693 1e-55 87
288 1 700475377H1 SATMON025 g450548 BLASTN 705 1e-55 90 289 1
700029625H1 SATMON003 g450548 BLASTN 712 1e-55 88 290 1 700202091H1
SATMON003 g1778820 BLASTN 774 1e-55 90 291 1 700264594H1 SATMON017
g450548 BLASTN 775 1e-55 89 292 1 700074969H1 SATMON007 g450548
BLASTN 777 1e-55 87 293 1 LIB3068-005-Q1-K1-A9 LIB3068 g1778820
BLASTN 389 1e-54 80 294 1 700207147H1 SATMON017 g1778820 BLASTN 537
1e-54 88 295 1 LIB189-004-Q1-E1-F11 LIB189 g960356 BLASTN 757 1e-54
87 296 1 700171222H1 SATMON013 g1778820 BLASTN 757 1e-54 93 297 1
700239903H1 SATMON010 g1778820 BLASTN 760 1e-54 84 298 1
700048208H1 SATMON003 g1778820 BLASTN 762 1e-54 90 299 1
700264085H1 SATMON017 g450548 BLASTN 764 1e-54 87 300 1 700155761H1
SATMON007 g1778820 BLASTN 579 1e-53 85 301 1 700216516H1 SATMON016
g450548 BLASTN 380 1e-52 89 302 1 LIB3068-044-Q1-K1-F12 LIB3068
g450548 BLASTN 492 1e-52 73 303 1 700344420H1 SATMON021 g450548
BLASTN 534 1e-52 79 304 1 700219767H1 SATMON011 g450548 BLASTN 732
1e-52 88 305 1 700153757H1 SATMON007 g1778820 BLASTN 737 1e-52 89
306 1 700165841H1 SATMON013 g450548 BLASTN 738 1e-52 90 307 1
700381966H1 SATMON023 g1778820 BLASTN 740 1e-52 86 308 1
700170427H1 SATMON013 g450548 BLASTN 687 1e-51 89 309 1 700094344H1
SATMON008 g1778820 BLASTN 725 1e-51 93 310 1 700052248H1 SATMON003
g1778820 BLASTN 726 1e-51 84 311 1 700050628H1 SATMON003 g450548
BLASTN 726 1e-51 89 312 1 700223031H1 SATMON011 g1778820 BLASTN 729
1e-51 89 313 1 700475686H1 SATMON025 g450548 BLASTN 660 1e-50 89
314 1 700170325H1 SATMON013 g2305013 BLASTN 709 1e-50 82 315 1
700221107H1 SATMON011 g450548 BLASTN 698 1e-49 89 316 1 700612589H1
SATMON033 g960356 BLASTN 703 1e-49 89 317 1 700153770H1 SATMON007
g1778820 BLASTN 705 1e-49 88 318 1 LIB3078-006-Q1-K1-E7 LIB3078
g1778820 BLASTN 705 1e-49 79 319 1 700239724H1 SATMON010 g1778820
BLASTN 507 1e-48 78 320 1 700356777H1 SATMON024 g1778820 BLASTN 683
1e-48 86 321 1 700241568H1 SATMON010 g960356 BLASTN 691 1e-48 88
322 1 700051294H1 SATMON003 g1778820 BLASTN 462 1e-47 88 323 1
700343661H1 SATMON021 g450548 BLASTN 507 1e-47 81 324 1 700029419H1
SATMON003 g1778820 BLASTN 547 1e-47 87 325 1 700165936H1 SATMON013
g1778820 BLASTN 671 1e-47 83 326 1 700026152H1 SATMON003 g960356
BLASTN 673 1e-47 89 327 1 700075671H1 SATMON007 g450548 BLASTN 429
1e-46 89 328 1 700156674H1 SATMON012 g1778820 BLASTN 659 1e-46 91
329 1 700094981H1 SATMON008 g1778820 BLASTN 662 1e-46 85 330 1
700092879H1 SATMON008 g1778820 BLASTN 668 1e-46 87 331 1
700240685H1 SATMON010 g1778820 BLASTN 484 1e-45 84 332 1
700150286H1 SATMON007 g1778820 BLASTN 647 1e-45 82 333 1
700104990H1 SATMON010 g450548 BLASTN 652 1e-45 89 334 1 700203829H1
SATMON003 g450548 BLASTN 652 1e-45 87 335 1 700153718H1 SATMON007
g167961 BLASTN 656 1e-45 91 336 1 700050841H1 SATMON003 g450548
BLASTN 571 1e-44 86 337 1 700268037H1 SATMON017 g2305013 BLASTN 636
1e-44 86 338 1 700153630H1 SATMON007 g1778820 BLASTN 637 1e-44 82
339 1 700475317H1 SATMON025 g450548 BLASTN 530 1e-43 87 340 1
701163127H1 SATMONN04 g960356 BLASTN 626 1e-43 88 341 1 700203688H1
SATMON003 g960356 BLASTN 627 1e-43 89 342 1 700049893H1 SATMON003
g1778820 BLASTN 506 1e-42 90 343 1 700449155H1 SATMON028 g1778820
BLASTN 443 1e-41 85 344 1 701183780H1 SATMONN06 g450548 BLASTN 558
1e-41 86 345 1 700242162H1 SATMON010 g1778820 BLASTN 600 1e-41 90
346 1 700466994H1 SATMON025 g450548 BLASTN 604 1e-41 91 347 1
700259823H1 SATMON017 g450548 BLASTN 607 1e-41 83 348 1 700346242H1
SATMON021 g450548 BLASTN 608 1e-41 86 349 1 700156395H1 SATMON007
g1778820 BLASTN 609 1e-41 90 350 1 700236835H1 SATMON010 g1778820
BLASTN 335 1e-40 81 351 1 700172385H1 SATMON013 g1778820 BLASTN 397
1e-40 87 352 1 700210466H1 SATMON016 g1778820 BLASTN 586 1e-40 81
353 1 700257303H1 SATMON017 g1778820 BLASTN 589 1e-40 81 354 1
LIB3067-027-Q1-K1-G1 LIB3067 g1778820 BLASTN 623 1e-40 87 355 1
LIB3066-025-Q1-K1-D1 LIB3066 g450548 BLASTN 524 1e-39 85 356 1
700106853H1 SATMON010 g450548 BLASTN 580 1e-39 86 357 1 700160540H1
SATMON012 g960356 BLASTN 581 1e-39 88 358 1 700157780H1 SATMON012
g960356 BLASTN 581 1e-39 88 359 1 700149801H1 SATMON007 g450548
BLASTN 565 1e-38 86 360 1 700353243H1 SATMON024 g450548 BLASTN 570
1e-38 86 361 1 700166171H1 SATMON013 g1778820 BLASTN 254 1e-37 79
362 1 700142509H1 SATMON012 g960356 BLASTN 556 1e-37 88 363 1
700242131H1 SATMON010 g450548 BLASTN 560 1e-37 85 364 1 700455643H1
SATMON029 g450548 BLASTN 539 1e-36 86 365 1 700150248H1 SATMON007
g1778820 BLASTN 547 1e-36 87 366 1 700208549H1 SATMON016 g450548
BLASTN 559 1e-36 88 367 1 700027193H1 SATMON003 g450548 BLASTN 529
1e-35 86 368 1 700221390H1 SATMON011 g450548 BLASTN 530 1e-35 85
369 1 700455647H1 SATMON029 g450548 BLASTN 531 1e-35 85 370 1
700260103H1 SATMON017 g1778820 BLASTN 531 1e-35 80 371 1
700167344H1 SATMON013 g1778820 BLASTN 536 1e-35 88 372 1
700089913H1 SATMON011 g960356 BLASTN 546 1e-35 88 373 1 700570573H1
SATMON030 g450548 BLASTN 300 1e-34 89 374 1 700169889H1 SATMON013
g1778820 BLASTN 519 1e-34 89 375 1 700262857H1 SATMON017 g1778820
BLASTN 521 1e-34 87 376 1 700142644H2 SATMON013 g1778820 BLASTN 522
1e-34 87 377 1 700224417H1 SATMON011 g450548 BLASTN 522 1e-34 91
378 1 700073882H1 SATMON007 g450548 BLASTN 531 1e-34 87 379 1
700085803H1 SATMON011 g450548 BLASTN 338 1e-33 89 380 1 700162323H1
SATMON012 g1778820 BLASTN 513 1e-33 86 381 1 700468306H1 SATMON025
g450548 BLASTN 519 1e-33 89 382 1 700443224H2 SATMON026 g450548
BLASTN 275 1e-32 83 383 1 LIB3066-054-Q1-K1-E3 LIB3066 g450548
BLASTN 495 1e-32 87 384 1 700211827H1 SATMON016 g1778820 BLASTN 489
1e-31 80 385 1 700048741H1 SATMON003 g450548 BLASTN 500 1e-31 88
386 1 700613620H1 SATMON033 g1778820 BLASTN 385 1e-30 88 387 1
700029203H1 SATMON003 g1778820 BLASTN 461 1e-29 84 388 1
700378431H1 SATMON020 g450548 BLASTN 466 1e-29 89 389 1 700455641H1
SATMON029 g450548 BLASTN 472 1e-29 85 390 1 700447867H1 SATMON027
g960356 BLASTN 473 1e-29 88 391 1 700447511H1 SATMON027 g450548
BLASTN 474 1e-29 88 392 1 700025851H1 SATMON003 g450548 BLASTN 479
1e-29 88 393 1 LIB3066-024-Q1-K1-H4 LIB3066 g1778820 BLASTN 481
1e-29 81 394 1 LIB143-025-Q1-E1-C4 LIB143 g450549 BLASTX 64 1e-28
70 395 1 700242282H1 SATMON010 g1778820 BLASTN 280 1e-28 85 396 1
700087244H1 SATMON011 g450548 BLASTN 464 1e-28 88 397 1 700458127H1
SATMON029 g450548 BLASTN 238 1e-27 82 398 1 700025767H1 SATMON003
g1778820 BLASTN 438 1e-26 86 399 1 700235401H1 SATMON010 g450548
BLASTN 442 1e-26 88 400 1 700029457H1 SATMON003 g450548 BLASTN 446
1e-26 89 401 1 700266174H1 SATMON017 g450548 BLASTN 447 1e-26 89
402 1 700349254H1 SATMON023 g1778820 BLASTN 315 1e-25 85 403 1
LIB3068-041-Q1-K1-H6 LIB3068 g450548 BLASTN 438 1e-25 86 404 1
700092889H1 SATMON008 g1778820 BLASTN 397 1e-24 92 405 1
700049044H1 SATMON003 g1778820 BLASTN 425 1e-24 90 406 1
700202710H1 SATMON003 g960357 BLASTX 114 1e-19 97 407 1 700155117H1
SATMON007 g450548 BLASTN 314 1e-19 90 408 1 700449619H1 SATMON028
g1778820 BLASTN 341 1e-19 89 409 1 700150336H1 SATMON007 g1778820
BLASTN 343 1e-19 87 410 1 700166223H1 SATMON013 g960357 BLASTX 181
1e-18 97 411 1 700153796H1 SATMON007 g2305013 BLASTN 300 1e-16 82
412 1 700053266H1 SATMON008 g450548 BLASTN 292 1e-15 83 413 1
700405227H1 SATMON028 g450548 BLASTN 313 1e-15 90 414 1 700397410H1
SATMONN01 g17262 BLASTX 147 1e-14 96 415 1 700211524H1 SATMON016
g1033190 BLASTX 153 1e-14 100 416 1 700281415H2 SATMON019 g2315140
BLASTX 139 1e-13 89 417 1 700429873H1 SATMONN01 g16961 BLASTX 133
1e-12 94 418 1 700239660H1 SATMON010 g450548 BLASTN 245 1e-12 86
419 1 700213596H1 SATMON016 g450548 BLASTN 258 1e-12 95 420 1
700152367H1 SATMON007 g450548 BLASTN 273 1e-12 88 421 1 700452014H1
SATMON028 g17262 BLASTX 76 1e-11 72 422 1 700357106H1 SATMON024
g1724104 BLASTX 132 1e-11 100 423 1 700468611H1 SATMON025 g450549
BLASTX 93 1e-10 93 424 1 700213526H1 SATMON016 g450549 BLASTX 127
1e-10 96 425 1 700267065H1 SATMON017 g450549 BLASTX 130 1e-10 96
426 1 700152044H1 SATMON007 g450549 BLASTX 88 1e-8 93 427 1
700159090H1 SATMON012 g169665 BLASTX 113 1e-8 81 428 1 700266734H1
SATMON017 g450549 BLASTX 119 1e-8 96 429 1 700405367H1 SATMON029
g1778820 BLASTN 231 1e-8 65 1635 -700555532 700555532H1 SOYMON001
g429103 BLASTN 920 1e-67 84 1636 -700649594 700649594H1 SOYMON003
g609559 BLASTX 186 1e-23 85 1637 -700750590 700750590H1 SOYMON014
g609224 BLASTN 363 1e-40 79 1638 -700755802 700755802H1 SOYMON014
g609224 BLASTN 479 1e-43 74 1639 -700869211 700869211H1 SOYMON016
g169665 BLASTX 146 1e-13 92 1640 -700891960 700891960H1 SOYMON024
g726031 BLASTN 589 1e-56 82 1641 -700900377 700900377H1 SOYMON027
g1655577 BLASTN 235 1e-8 78 1642 -700902427 700902427H1 SOYMON027
g1655576 BLASTX 151 1e-13 76 1643 -700941686 700941686H1 SOYMON024
g497899 BLASTN 442 1e-26 89 1644 -700952418 700952418H1 SOYMON022
g726030 BLASTX 146 1e-17 83 1645 -700979651 700979651H2 SOYMON009
g1655579 BLASTN 1006 1e-75 84 1646 -700982809 700982809H1 SOYMON009
g1655579 BLASTN 945 1e-69 82 1647 -700982867 700982867H1 SOYMON009
g1127582 BLASTN 868 1e-63 78 1648 -701056884 701056884H1 SOYMON032
g609556 BLASTN 726 1e-51 84 1649 -701117318 701117318H1 SOYMON037
g609224 BLASTN 451 1e-37 80 1650 -701118224 701118224H1 SOYMON037
g2305013 BLASTN 285 1e-19 72 1651 -701121264 701121264H1 SOYMON037
g166873 BLASTN 238 1e-8 82 1652 -701122908 701122908H1 SOYMON037
g16508 BLASTN 444 1e-26 83 1653 -701128589 701128589H1 SOYMON037
g16508 BLASTN 539 1e-36 90 1654 -GM12798 LIB3049-039-Q1-E1-F2
LIB3049 g16508 BLASTN 578 1e-37 73 1655 -GM14331
LIB3049-055-Q1-E1-F5 LIB3049 g167961 BLASTN 497 1e-30 62 1656
-GM30881 LIB3050-005-Q1-K1-G1 LIB3050 g1655577 BLASTN 543 1e-34 82
1657 -GM30911 LIB3050-005-Q1-K1-B8 LIB3050 g1655577 BLASTN 387
1e-26 71 1658 -GM33921 LIB3051-028-Q1-K1-A9 LIB3051 g16508 BLASTN
338 1e-32 73 1659 12644 701131794H1 SOYMON038 g429107 BLASTN 877
1e-64 84 1660 12644 701142515H1 SOYMON038 g429107 BLASTN 871 1e-63
84 1661 12644 700888494H1 SOYMON024 g429107 BLASTN 662 1e-46 83
1662 16 LIB3030-009-Q1-B1-C1 LIB3030 g429105 BLASTN 1439 1e-119 84
1663 16 LIB3051-106-Q1-K1-B5 LIB3051 g609224 BLASTN 1516 1e-117 86
1664 16 LIB3050-023-Q1-K1-A12 LIB3050 g1724103 BLASTN 1388 1e-106
83 1665 16 LIB3028-003-Q1-B1-G11 LIB3028 g609224 BLASTN 1313 1e-100
87 1666 16 LIB3054-010-Q1-N1-E2 LIB3054 g609224 BLASTN 920 1e-95 87
1667 16 700651294H1 SOYMON003 g609224 BLASTN 672 1e-94 86 1668 16
LIB3027-007-Q1-B1-G3 LIB3027 g16508 BLASTN 891 1e-93 83 1669 16
LIB3053-013-Q1-N1-H9 LIB3053 g609224 BLASTN 996 1e-93 87 1670 16
LIB3051-061-Q1-K1-C7 LIB3051 g16508 BLASTN 1195 1e-93 86 1671 16
LIB3065-005-Q1-N1-A6 LIB3065 g609224 BLASTN 707 1e-91 78 1672 16
LIB3050-008-Q1-E1-E3 LIB3050 g609224 BLASTN 1102 1e-87 87 1673 16
LIB3051-011-Q1-E1-B2 LIB3051 g16508 BLASTN 1153 1e-87 82 1674 16
LIB3030-010-Q1-B1-F8 LIB3030 g609224 BLASTN 988 1e-86 83 1675 16
700652104H1 SOYMON003 g1655577 BLASTN 990 1e-86 83 1676 16
701205279H1 SOYMON035 g609556 BLASTN 1125 1e-85 86 1677 16
700662183H1 SOYMON005 g609224 BLASTN 678 1e-83 84 1678 16
LIB3051-039-Q1-K1-F5 LIB3051 g169664 BLASTN 1104 1e-83 86 1679 16
700557616H1 SOYMON001 g609556 BLASTN 1111 1e-83 86 1680 16
LIB3030-002-Q1-B1-C6 LIB3030 g16508 BLASTN 1113 1e-83 83 1681 16
700865235H1 SOYMON016 g1724103 BLASTN 1090 1e-82 84 1682 16
700563340H1 SOYMON002 g609224 BLASTN 911 1e-80 86 1683 16
LIB3040-044-Q1-E1-D7 LIB3040 g166873 BLASTN 1052 1e-80 82 1684 16
LIB3040-060-Q1-E1-D9 LIB3040 g609224 BLASTN 1071 1e-80 84 1685 16
LIB3049-001-Q1-E1-F12 LIB3049 g16508 BLASTN 1074 1e-80 84 1686 16
LIB3028-006-Q1-B1-F1 LIB3028 g16508 BLASTN 857 1e-79 83 1687 16
LIB3049-033-Q1-E1-G5 LIB3049 g2315139 BLASTN 933 1e-79 81 1688 16
700978240H1 SOYMON009 g609224 BLASTN 984 1e-79 88 1689 16
700980802H1 SOYMON009 g862999 BLASTN 1060 1e-79 85 1690 16
700755908H1 SOYMON014 g1724103 BLASTN 1062 1e-79 88 1691 16
700562226H1 SOYMON002 g1724103 BLASTN 1065 1e-79 84 1692 16
701119101H1 SOYMON037 g609224 BLASTN 1048 1e-78 86 1693 16
700646291H1 SOYMON012 g1655577 BLASTN 1049 1e-78 86 1694 16
LIB3050-022-Q1-K1-B9 LIB3050 g16508 BLASTN 1049 1e-78 83 1695 16
LIB3028-030-Q1-B1-F8 LIB3028 g16508 BLASTN 1053 1e-78 83 1696 16
701143128H1 SOYMON038 g609556 BLASTN 737 1e-77 86 1697 16
LIB3040-041-Q1-E1-D10 LIB3040 g166873 BLASTN 861 1e-77 81 1698 16
700756363H1 SOYMON014 g609556 BLASTN 1031 1e-77 88 1699 16
700729963H1 SOYMON009 g1724103 BLASTN 1036 1e-77 88
1700 16 701063046H1 SOYMON033 g609224 BLASTN 1037 1e-77 86 1701 16
LIB3030-005-Q1-B1-G2 LIB3030 g609224 BLASTN 1038 1e-77 84 1702 16
700564331H1 SOYMON002 g609556 BLASTN 1039 1e-77 84 1703 16
700562958H1 SOYMON002 g1724103 BLASTN 1040 1e-77 88 1704 16
LIB3039-014-Q1-E1-F7 LIB3039 g16508 BLASTN 569 1e-76 84 1705 16
700753632H1 SOYMON014 g497899 BLASTN 668 1e-76 83 1706 16
LIB3049-048-Q1-E1-F2 LIB3049 g16508 BLASTN 722 1e-76 83 1707 16
700648911H1 SOYMON003 g1655577 BLASTN 983 1e-76 83 1708 16
LIB3040-027-Q1-E1-H3 LIB3040 g166873 BLASTN 1003 1e-76 81 1709 16
700664905H1 SOYMON005 g609556 BLASTN 1020 1e-76 86 1710 16
701133404H1 SOYMON038 g1724103 BLASTN 1023 1e-76 85 1711 16
701208780H1 SOYMON035 g1655577 BLASTN 1025 1e-76 85 1712 16
700902279H1 SOYMON027 g169664 BLASTN 1026 1e-76 88 1713 16
LIB3040-048-Q1-E1-G10 LIB3040 g16508 BLASTN 1030 1e-76 83 1714 16
LIB3040-028-Q1-E1-A4 LIB3040 g16508 BLASTN 979 1e-75 84 1715 16
700807586H1 SOYMON016 g609224 BLASTN 1006 1e-75 86 1716 16
700755486H1 SOYMON014 g609224 BLASTN 1006 1e-75 87 1717 16
700724920H1 SOYMON009 g609224 BLASTN 1009 1e-75 87 1718 16
701007494H2 SOYMON019 g1724103 BLASTN 1013 1e-75 85 1719 16
700568310H1 SOYMON002 g497899 BLASTN 834 1e-74 83 1720 16
701208819H1 SOYMON035 g609224 BLASTN 857 1e-74 87 1721 16
700985084H1 SOYMON009 g609224 BLASTN 872 1e-74 85 1722 16
701013074H1 SOYMON019 g726031 BLASTN 894 1e-74 88 1723 16
700650843H1 SOYMON003 g609224 BLASTN 924 1e-74 86 1724 16
700646037H1 SOYMON011 g609556 BLASTN 930 1e-74 84 1725 16
LIB3040-039-Q1-E1-H9 LIB3040 g166873 BLASTN 973 1e-74 80 1726 16
700847231H1 SOYMON021 g726031 BLASTN 999 1e-74 85 1727 16
700792293H1 SOYMON011 g609224 BLASTN 999 1e-74 86 1728 16
701109644H1 SOYMON036 g609556 BLASTN 1004 1e-74 86 1729 16
701122929H1 SOYMON037 g1724103 BLASTN 459 1e-73 87 1730 16
700661491H1 SOYMON005 g16508 BLASTN 540 1e-73 90 1731 16
701120070H1 SOYMON037 g609224 BLASTN 786 1e-73 87 1732 16
700898895H1 SOYMON027 g609556 BLASTN 820 1e-73 88 1733 16
701096959H1 SOYMON028 g429105 BLASTN 839 1e-73 82 1734 16
700848356H1 SOYMON021 g497899 BLASTN 985 1e-73 85 1735 16
700864876H1 SOYMON016 g1724103 BLASTN 985 1e-73 88 1736 16
700868456H1 SOYMON016 g609224 BLASTN 987 1e-73 86 1737 16
701210488H1 SOYMON035 g1655577 BLASTN 988 1e-73 83 1738 16
700747764H1 SOYMON013 g609224 BLASTN 822 1e-72 87 1739 16
701063311H1 SOYMON033 g1655577 BLASTN 971 1e-72 86 1740 16
700868625H1 SOYMON016 g1724103 BLASTN 973 1e-72 85 1741 16
700992344H1 SOYMON011 g726031 BLASTN 973 1e-72 85 1742 16
701012719H1 SOYMON019 g1724103 BLASTN 973 1e-72 85 1743 16
700676769H1 SOYMON007 g609224 BLASTN 973 1e-72 87 1744 16
700946235H1 SOYMON024 g609224 BLASTN 975 1e-72 85 1745 16
700891218H1 SOYMON024 g1724103 BLASTN 975 1e-72 84 1746 16
700724907H1 SOYMON009 g609556 BLASTN 977 1e-72 84 1747 16
700833549H1 SOYMON019 g1655577 BLASTN 979 1e-72 85 1748 16
700942105H1 SOYMON024 g726031 BLASTN 980 1e-72 84 1749 16
700564290H1 SOYMON002 g726031 BLASTN 530 1e-71 84 1750 16
700891233H1 SOYMON024 g1724103 BLASTN 746 1e-71 86 1751 16
LIB3028-025-Q1-B1-B2 LIB3028 g609224 BLASTN 870 1e-71 85 1752 16
701120896H1 SOYMON037 g429105 BLASTN 959 1e-71 82 1753 16
LIB3051-027-Q1-K1-A9 LIB3051 g609224 BLASTN 962 1e-71 83 1754 16
701050696H1 SOYMON032 g609556 BLASTN 965 1e-71 86 1755 16
700653619H1 SOYMON003 g609224 BLASTN 966 1e-71 86 1756 16
701047994H1 SOYMON032 g1724103 BLASTN 967 1e-71 85 1757 16
701123056H1 SOYMON037 g1724103 BLASTN 968 1e-71 85 1758 16
700983479H1 SOYMON009 g726031 BLASTN 653 1e-70 85 1759 16
701063733H1 SOYMON034 g1655577 BLASTN 850 1e-70 85 1760 16
700738366H1 SOYMON012 g726031 BLASTN 946 1e-70 85 1761 16
700866319H1 SOYMON016 g1655575 BLASTN 947 1e-70 85 1762 16
700789013H2 SOYMON011 g1655577 BLASTN 948 1e-70 85 1763 16
700896080H1 SOYMON027 g609224 BLASTN 949 1e-70 87 1764 16
LIB3039-021-Q1-E1-F12 LIB3039 g16508 BLASTN 950 1e-70 83 1765 16
700898284H1 SOYMON027 g429105 BLASTN 950 1e-70 86 1766 16
700901743H1 SOYMON027 g609224 BLASTN 950 1e-70 86 1767 16
700831048H1 SOYMON019 g429105 BLASTN 951 1e-70 85 1768 16
701038107H1 SOYMON029 g726031 BLASTN 952 1e-70 87 1769 16
700559703H1 SOYMON001 g726031 BLASTN 954 1e-70 88 1770 16
700848817H1 SOYMON021 g609224 BLASTN 954 1e-70 84 1771 16
700896032H1 SOYMON027 g609224 BLASTN 956 1e-70 87 1772 16
700944764H1 SOYMON024 g16508 BLASTN 567 1e-69 85 1773 16
701011015H1 SOYMON019 g1724103 BLASTN 647 1e-69 81 1774 16
701125662H1 SOYMON037 g609224 BLASTN 751 1e-69 87 1775 16
701046895H1 SOYMON032 g16508 BLASTN 795 1e-69 83 1776 16
701012632H1 SOYMON019 g1655577 BLASTN 800 1e-69 85 1777 16
701005244H1 SOYMON019 g1724103 BLASTN 804 1e-69 82 1778 16
LIB3050-023-Q1-K1-H10 LIB3050 g609224 BLASTN 899 1e-69 83 1779 16
700892558H1 SOYMON024 g1655575 BLASTN 936 1e-69 85 1780 16
700988779H1 SOYMON011 g16508 BLASTN 937 1e-69 82 1781 16
701203923H2 SOYMON035 g429105 BLASTN 938 1e-69 86 1782 16
701010231H2 SOYMON019 g1724103 BLASTN 938 1e-69 83 1783 16
701041790H1 SOYMON029 g450548 BLASTN 939 1e-69 84 1784 16
700967887H1 SOYMON033 g1724103 BLASTN 940 1e-69 87 1785 16
701123361H1 SOYMON037 g16508 BLASTN 941 1e-69 84 1786 16
701123154H1 SOYMON037 g1724103 BLASTN 942 1e-69 86 1787 16
701045767H1 SOYMON032 g726031 BLASTN 942 1e-69 85 1788 16
700983288H1 SOYMON009 g609224 BLASTN 945 1e-69 83 1789 16
700653053H1 SOYMON003 g609224 BLASTN 945 1e-69 86 1790 16
701044226H1 SOYMON032 g1655577 BLASTN 486 1e-68 83 1791 16
700969927H1 SOYMON005 g497899 BLASTN 750 1e-68 85 1792 16
700547956H1 SOYMON001 g609224 BLASTN 779 1e-68 87 1793 16
LIB3049-048-Q1-E1-E4 LIB3049 g609224 BLASTN 827 1e-68 83 1794 16
701140780H1 SOYMON038 g1655577 BLASTN 922 1e-68 85 1795 16
700849166H1 SOYMON021 g1724103 BLASTN 923 1e-68 84 1796 16
700891945H1 SOYMON024 g609556 BLASTN 925 1e-68 87 1797 16
700658051H1 SOYMON004 g429105 BLASTN 925 1e-68 83 1798 16
700942415H1 SOYMON024 g1655575 BLASTN 928 1e-68 84 1799 16
701041890H1 SOYMON029 g1724103 BLASTN 930 1e-68 82 1800 16
LIB3028-006-Q1-B1-H12 LIB3028 g16508 BLASTN 931 1e-68 81 1801 16
700967039H1 SOYMON029 g429105 BLASTN 931 1e-68 85 1802 16
700836178H1 SOYMON019 g1724103 BLASTN 933 1e-68 85 1803 16
700897558H1 SOYMON027 g16508 BLASTN 737 1e-67 85 1804 16
LIB3050-004-Q1-E1-A2 LIB3050 g609224 BLASTN 796 1e-67 80 1805 16
700730236H1 SOYMON009 g726031 BLASTN 816 1e-67 84 1806 16
700833936H1 SOYMON019 g1724103 BLASTN 915 1e-67 84 1807 16
700897552H1 SOYMON027 g1655577 BLASTN 916 1e-67 84 1808 16
700945440H1 SOYMON024 g609556 BLASTN 917 1e-67 84 1809 16
700961368H1 SOYMON022 g169664 BLASTN 918 1e-67 88 1810 16
700789702H1 SOYMON011 g609224 BLASTN 921 1e-67 86 1811 16
700786541H1 SOYMON011 g429105 BLASTN 513 1e-66 84 1812 16
700987282H1 SOYMON009 g16508 BLASTN 523 1e-66 87 1813 16
700661002H1 SOYMON005 g2305013 BLASTN 836 1e-66 81 1814 16
701148312H1 SOYMON031 g862999 BLASTN 899 1e-66 84 1815 16
700738822H1 SOYMON012 g1655577 BLASTN 899 1e-66 85 1816 16
700940996H1 SOYMON024 g609556 BLASTN 901 1e-66 86 1817 16
700749195H1 SOYMON013 g609556 BLASTN 903 1e-66 81 1818 16
700893941H1 SOYMON024 g429105 BLASTN 903 1e-66 88 1819 16
700892888H1 SOYMON024 g609556 BLASTN 906 1e-66 86 1820 16
700901481H1 SOYMON027 g169664 BLASTN 638 1e-65 86 1821 16
700945269H1 SOYMON024 g169664 BLASTN 688 1e-65 86 1822 16
700746876H1 SOYMON013 g609556 BLASTN 740 1e-65 85 1823 16
700755043H1 SOYMON014 g609556 BLASTN 760 1e-65 87 1824 16
701097166H1 SOYMON028 g1655577 BLASTN 781 1e-65 82 1825 16
701129484H1 SOYMON037 g16508 BLASTN 898 1e-65 82 1826 16
700651014H1 SOYMON003 g167961 BLASTN 464 1e-64 81 1827 16
701134363H1 SOYMON038 g726031 BLASTN 687 1e-64 84 1828 16
701124677H1 SOYMON037 g726031 BLASTN 876 1e-64 85 1829 16
701000359H1 SOYMON018 g1724103 BLASTN 877 1e-64 87 1830 16
701139375H1 SOYMON038 g1724103 BLASTN 883 1e-64 84 1831 16
700832784H1 SOYMON019 g609224 BLASTN 883 1e-64 89 1832 16
700980227H1 SOYMON009 g1724103 BLASTN 884 1e-64 84 1833 16
700943177H1 SOYMON024 g1655577 BLASTN 885 1e-64 85 1834 16
700844446H1 SOYMON021 g2315139 BLASTN 355 1e-63 84 1835 16
700836453H1 SOYMON020 g862999 BLASTN 494 1e-63 85 1836 16
700983012H1 SOYMON009 g16508 BLASTN 756 1e-63 83 1837 16
700795805H1 SOYMON017 g1655577 BLASTN 833 1e-63 85 1838 16
701213663H1 SOYMON035 g1724103 BLASTN 862 1e-63 84 1839 16
700868366H1 SOYMON016 g167961 BLASTN 864 1e-63 83 1840 16
701134377H1 SOYMON038 g2305013 BLASTN 865 1e-63 79 1841 16
LIB3049-007-Q1-E1-C9 LIB3049 g16508 BLASTN 867 1e-63 79 1842 16
700944577H1 SOYMON024 g862999 BLASTN 869 1e-63 85 1843 16
700992289H1 SOYMON011 g16508 BLASTN 869 1e-63 84 1844 16
700891612H1 SOYMON024 g609556 BLASTN 872 1e-63 86 1845 16
700738277H1 SOYMON012 g609224 BLASTN 463 1e-62 85 1846 16
700895571H1 SOYMON027 g609224 BLASTN 562 1e-62 88 1847 16
LIB3049-008-Q1-E1-B3 LIB3049 g16508 BLASTN 853 1e-62 77 1848 16
LIB3027-010-Q1-B1-E12 LIB3027 g609224 BLASTN 854 1e-62 82 1849 16
701129389H1 SOYMON037 g16508 BLASTN 854 1e-62 83 1850 16
701045542H1 SOYMON032 g429105 BLASTN 855 1e-62 86 1851 16
700969901H1 SOYMON005 g726031 BLASTN 856 1e-62 84 1852 16
700984664H1 SOYMON009 g1724103 BLASTN 856 1e-62 81 1853 16
700561352H1 SOYMON002 g609224 BLASTN 858 1e-62 85 1854 16
701138828H1 SOYMON038 g16508 BLASTN 858 1e-62 84 1855 16
700845962H1 SOYMON021 g1655577 BLASTN 861 1e-62 87 1856 16
700896147H1 SOYMON027 g16508 BLASTN 336 1e-61 82 1857 16
701137929H1 SOYMON038 g1655577 BLASTN 502 1e-61 84 1858 16
LIB3040-032-Q1-E1-B8 LIB3040 g16508 BLASTN 521 1e-61 82 1859 16
701009537H1 SOYMON019 g726031 BLASTN 545 1e-61 86 1860 16
LIB3049-031-Q1-E1-E4 LIB3049 g609224 BLASTN 677 1e-61 82 1861 16
LIB3049-031-Q1-E1-C7 LIB3049 g167961 BLASTN 696 1e-61 82 1862 16
700891843H1 SOYMON024 g1655577 BLASTN 737 1e-61 84 1863 16
700561231H1 SOYMON002 g16508 BLASTN 839 1e-61 82 1864 16
700548238H1 SOYMON002 g429105 BLASTN 843 1e-61 85 1865 16
700901018H1 SOYMON027 g2305013 BLASTN 845 1e-61 82 1866 16
701134413H1 SOYMON038 g2305013 BLASTN 848 1e-61 82 1867 16
700653524H1 SOYMON003 g609224 BLASTN 499 1e-60 85 1868 16
LIB3049-017-Q1-E1-G8 LIB3049 g16508 BLASTN 514 1e-60 82 1869 16
701121077H1 SOYMON037 g16508 BLASTN 671 1e-60 84 1870 16
LIB3056-013-Q1-N1-A9 LIB3056 g609224 BLASTN 784 1e-60 86 1871 16
700943524H1 SOYMON024 g16508 BLASTN 831 1e-60 85 1872 16
700952889H1 SOYMON022 g429103 BLASTN 835 1e-60 84 1873 16
700730632H1 SOYMON009 g1655577 BLASTN 431 1e-59 82 1874 16
700649136H1 SOYMON003 g609224 BLASTN 486 1e-59 84 1875 16
700895591H1 SOYMON027 g609556 BLASTN 492 1e-59 87 1876 16
701009829H1 SOYMON019 g429103 BLASTN 572 1e-59 80 1877 16
LIB3039-011-Q1-E1-C11 LIB3039 g16508 BLASTN 619 1e-59 83 1878 16
LIB3040-055-Q1-E1-C4 LIB3040 g16508 BLASTN 622 1e-59 82 1879 16
701123571H1 SOYMON037 g2305013 BLASTN 682 1e-59 81 1880 16
LIB3049-004-Q1-E1-E5 LIB3049 g167961 BLASTN 771 1e-59 83 1881 16
701002239H1 SOYMON018 g609224 BLASTN 782 1e-59 86 1882 16
700971088H1 SOYMON005 g1724103 BLASTN 793 1e-59 81 1883 16
700864624H1 SOYMON016 g167961 BLASTN 817 1e-59 83 1884 16
700863534H1 SOYMON027 g16508 BLASTN 818 1e-59 86 1885 16
700749489H1 SOYMON013 g16508 BLASTN 823 1e-59 86 1886 16
700730689H1 SOYMON009 g16508 BLASTN 825 1e-59 83 1887 16
LIB3049-050-Q1-E1-A11 LIB3049 g16508 BLASTN 831 1e-59 83 1888 16
700850803H1 SOYMON023 g1655577 BLASTN 511 1e-58 83 1889 16
700992317H1 SOYMON011 g1655577 BLASTN 520 1e-58 79 1890 16
701119129H1 SOYMON037 g16508 BLASTN 553 1e-58 83 1891 16
700657623H1 SOYMON004 g1724103 BLASTN 600 1e-58 83 1892 16
LIB3049-015-Q1-E1-F8 LIB3049 g16508 BLASTN 674 1e-58 79 1893 16
701007027H1 SOYMON019 g16508 BLASTN 804 1e-58 81 1894 16
701120820H1 SOYMON037 g609224 BLASTN 807 1e-58 85 1895 16
700654317H1 SOYMON004 g1655577 BLASTN 811 1e-58 84 1896 16
700889071H1 SOYMON024 g497899 BLASTN 812 1e-58 83 1897 16
701010676H1 SOYMON019 g609224 BLASTN 813 1e-58 85 1898 16
701099590H1 SOYMON028 g1655577 BLASTN 486 1e-57 82 1899 16
700649684H1 SOYMON003 g16508 BLASTN 540 1e-57 82 1900 16
700994107H1 SOYMON011 g16508 BLASTN 567 1e-57 80 1901 16
700898629H1 SOYMON027 g16508 BLASTN 687 1e-57 82 1902 16
700902333H1 SOYMON027 g609224 BLASTN 700 1e-57 82 1903 16
LIB3039-017-Q1-E1-D9 LIB3039 g16508 BLASTN 703 1e-57 81 1904 16
LIB3040-026-Q1-E1-A4 LIB3040 g16508 BLASTN 709 1e-57 83 1905 16
701130101H1 SOYMON037 g1724103 BLASTN 791 1e-57 88 1906 16
700902482H1 SOYMON027 g169664 BLASTN 797 1e-57 81 1907 16
700730789H1 SOYMON009 g16508 BLASTN 798 1e-57 82 1908 16
700868670H1 SOYMON016 g16508 BLASTN 800 1e-57 83 1909 16
LIB3029-011-Q1-B1-D1 LIB3029 g609224 BLASTN 800 1e-57 82 1910 16
701119278H1 SOYMON037 g609224 BLASTN 801 1e-57 83 1911 16
700747731H1 SOYMON013 g16508 BLASTN 801 1e-57 86 1912 16
LIB3049-017-Q1-E1-A11 LIB3049 g16508 BLASTN 811 1e-57 80 1913 16
700890428H1 SOYMON024 g609224 BLASTN 482 1e-56 83 1914 16
700562326H1 SOYMON002 g609224 BLASTN 778 1e-56 82 1915 16
700567740H1 SOYMON002 g609224 BLASTN 781 1e-56 85 1916 16
701061514H1 SOYMON033 g1724103 BLASTN 784 1e-56 86 1917 16
701135670H1 SOYMON038 g429105 BLASTN 785 1e-56 81 1918 16
701139657H1 SOYMON038 g609224 BLASTN 786 1e-56 85 1919 16
701123371H1 SOYMON037 g16508 BLASTN 788 1e-56 86 1920 16
700838744H1 SOYMON020 g16508 BLASTN 789 1e-56 82 1921 16
701070458H1 SOYMON034 g1655577 BLASTN 457 1e-55 78 1922 16
701003437H1 SOYMON019 g429105 BLASTN 474 1e-55 83 1923 16
700835976H1 SOYMON019 g16508 BLASTN 562 1e-55 84 1924 16
700894535H1 SOYMON024 g16508 BLASTN 570 1e-55 86 1925 16
700752755H1 SOYMON014 g1655577 BLASTN 690 1e-55 83 1926 16
LIB3049-032-Q1-E1-C9 LIB3049 g16508 BLASTN 700 1e-55 83 1927 16
LIB3040-030-Q1-E1-B5 LIB3040 g16508 BLASTN 709 1e-55 84 1928 16
701136935H1 SOYMON038 g609224 BLASTN 766 1e-55 85 1929 16
700682088H1 SOYMON008 g169664 BLASTN 767 1e-55 90 1930 16
700682188H1 SOYMON008 g169664 BLASTN 767 1e-55 90 1931 16
701068649H1 SOYMON034 g16508 BLASTN 770 1e-55 83 1932 16
700726623H1 SOYMON009 g16508 BLASTN 771 1e-55 85 1933 16
700751129H1 SOYMON014 g16508 BLASTN 772 1e-55 86 1934 16
700945234H1 SOYMON024 g16508 BLASTN 773 1e-55 83 1935 16
701133507H2 SOYMON038 g609224 BLASTN 776 1e-55 85 1936 16
700902459H1 SOYMON027 g726031 BLASTN 777 1e-55 84 1937 16
700986624H1 SOYMON009 g16508 BLASTN 612 1e-54 83 1938 16
701097020H1 SOYMON028 g609224 BLASTN 615 1e-54 84 1939 16
701131409H1 SOYMON038 g16508 BLASTN 755 1e-54 91 1940 16
700750123H1 SOYMON013 g726031 BLASTN 756 1e-54 86 1941 16
701040251H1 SOYMON029 g16508 BLASTN 756 1e-54 85 1942 16
700732290H1 SOYMON010 g169664 BLASTN 758 1e-54 90 1943 16
701117458H1 SOYMON037 g609224 BLASTN 759 1e-54 82 1944 16
701118709H1 SOYMON037 g16508 BLASTN 760 1e-54 91 1945 16
701012834H1 SOYMON019 g16508 BLASTN 760 1e-54 91 1946 16
701133316H1 SOYMON038 g16508 BLASTN 760 1e-54 91 1947 16
701129646H1 SOYMON037 g16508 BLASTN 760 1e-54 91 1948 16
700732265H1 SOYMON010 g169664 BLASTN 760 1e-54 90 1949 16
701040796H1 SOYMON029 g1724103 BLASTN 760 1e-54 89 1950 16
700846350H1 SOYMON021 g16508 BLASTN 761 1e-54 86
1951 16 701137005H1 SOYMON038 g16508 BLASTN 761 1e-54 86 1952 16
701046506H1 SOYMON032 g16508 BLASTN 761 1e-54 91 1953 16
700894462H1 SOYMON024 g16508 BLASTN 761 1e-54 86 1954 16
700973847H1 SOYMON005 g16508 BLASTN 766 1e-54 87 1955 16
701128215H1 SOYMON037 g609556 BLASTN 466 1e-53 82 1956 16
700842468H1 SOYMON020 g429105 BLASTN 548 1e-53 86 1957 16
701051552H1 SOYMON032 g609224 BLASTN 744 1e-53 85 1958 16
700944049H1 SOYMON024 g609224 BLASTN 744 1e-53 85 1959 16
701120261H1 SOYMON037 g609224 BLASTN 746 1e-53 84 1960 16
700897524H1 SOYMON027 g609224 BLASTN 751 1e-53 84 1961 16
LIB3049-042-Q1-E1-F7 LIB3049 g450548 BLASTN 761 1e-53 75 1962 16
700896909H1 SOYMON027 g429105 BLASTN 536 1e-52 82 1963 16
700974820H1 SOYMON005 g1724103 BLASTN 582 1e-52 86 1964 16
701143224H1 SOYMON038 g609224 BLASTN 731 1e-52 84 1965 16
701010322H1 SOYMON019 g609224 BLASTN 731 1e-52 85 1966 16
701130510H1 SOYMON038 g609224 BLASTN 732 1e-52 83 1967 16
700724904H1 SOYMON009 g609224 BLASTN 733 1e-52 85 1968 16
701119845H1 SOYMON037 g609224 BLASTN 734 1e-52 85 1969 16
701107871H1 SOYMON036 g16508 BLASTN 737 1e-52 86 1970 16
700983380H1 SOYMON009 g609224 BLASTN 738 1e-52 84 1971 16
700896469H1 SOYMON027 g16508 BLASTN 741 1e-52 86 1972 16
700792178H1 SOYMON011 g16508 BLASTN 742 1e-52 85 1973 16
701099906H1 SOYMON028 g16508 BLASTN 431 1e-51 89 1974 16
701134724H2 SOYMON038 g16508 BLASTN 553 1e-51 85 1975 16
700749712H1 SOYMON013 g16508 BLASTN 689 1e-51 87 1976 16
700982567H1 SOYMON009 g609224 BLASTN 723 1e-51 85 1977 16
701013758H1 SOYMON019 g609224 BLASTN 723 1e-51 85 1978 16
700868508H1 SOYMON016 g609224 BLASTN 723 1e-51 85 1979 16
700749316H1 SOYMON013 g609224 BLASTN 723 1e-51 85 1980 16
701045367H1 SOYMON032 g16508 BLASTN 724 1e-51 91 1981 16
701205494H1 SOYMON035 g609224 BLASTN 724 1e-51 84 1982 16
700556784H1 SOYMON001 g16508 BLASTN 724 1e-51 91 1983 16
701099879H1 SOYMON028 g16508 BLASTN 726 1e-51 80 1984 16
701131671H1 SOYMON038 g609224 BLASTN 728 1e-51 85 1985 16
701011505H1 SOYMON019 g609224 BLASTN 728 1e-51 85 1986 16
701009973H2 SOYMON019 g609224 BLASTN 728 1e-51 85 1987 16
700793520H1 SOYMON017 g609224 BLASTN 728 1e-51 85 1988 16
700753622H1 SOYMON014 g609224 BLASTN 728 1e-51 85 1989 16
700984052H1 SOYMON009 g609224 BLASTN 728 1e-51 83 1990 16
701108521H1 SOYMON036 g609224 BLASTN 728 1e-51 85 1991 16
700957203H1 SOYMON022 g1724103 BLASTN 464 1e-50 86 1992 16
700554120H1 SOYMON001 g609224 BLASTN 528 1e-50 84 1993 16
700555954H1 SOYMON001 g167961 BLASTN 528 1e-50 86 1994 16
701124141H1 SOYMON037 g609224 BLASTN 537 1e-50 85 1995 16
701003232H1 SOYMON019 g16508 BLASTN 544 1e-50 88 1996 16
700653112H1 SOYMON003 g16508 BLASTN 559 1e-50 91 1997 16
701103353H1 SOYMON028 g1655577 BLASTN 581 1e-50 84 1998 16
700562790H1 SOYMON002 g497899 BLASTN 708 1e-50 86 1999 16
701097303H1 SOYMON028 g609224 BLASTN 711 1e-50 84 2000 16
700561195H1 SOYMON002 g609224 BLASTN 712 1e-50 84 2001 16
701121162H1 SOYMON037 g16508 BLASTN 712 1e-50 88 2002 16
700981317H1 SOYMON009 g16508 BLASTN 714 1e-50 89 2003 16
700981595H1 SOYMON009 g16508 BLASTN 716 1e-50 86 2004 16
700994305H1 SOYMON011 g609224 BLASTN 528 1e-49 85 2005 16
701101565H1 SOYMON028 g429105 BLASTN 552 1e-49 82 2006 16
701103456H1 SOYMON028 g429105 BLASTN 617 1e-49 81 2007 16
700993331H1 SOYMON011 g16508 BLASTN 622 1e-49 86 2008 16
LIB3052-011-Q1-N1-B12 LIB3052 g16508 BLASTN 695 1e-49 85 2009 16
701140613H1 SOYMON038 g609224 BLASTN 697 1e-49 85 2010 16
700745426H1 SOYMON013 g16508 BLASTN 697 1e-49 84 2011 16
701046530H1 SOYMON032 g609224 BLASTN 697 1e-49 84 2012 16
701003787H1 SOYMON019 g609224 BLASTN 697 1e-49 85 2013 16
700840830H1 SOYMON020 g1724103 BLASTN 699 1e-49 79 2014 16
700901588H1 SOYMON027 g609224 BLASTN 702 1e-49 85 2015 16
701037824H1 SOYMON029 g497899 BLASTN 702 1e-49 87 2016 16
701131888H1 SOYMON038 g609224 BLASTN 702 1e-49 85 2017 16
701009947H2 SOYMON019 g609224 BLASTN 702 1e-49 85 2018 16
700729216H1 SOYMON009 g497899 BLASTN 702 1e-49 87 2019 16
700831705H1 SOYMON019 g609224 BLASTN 702 1e-49 85 2020 16
700900329H1 SOYMON027 g429105 BLASTN 705 1e-49 88 2021 16
700563946H1 SOYMON002 g16508 BLASTN 705 1e-49 83 2022 16
700808370H1 SOYMON024 g609556 BLASTN 459 1e-48 83 2023 16
LIB3040-001-Q1-E1-H4 LIB3040 g1724103 BLASTN 475 1e-48 76 2024 16
700989482H1 SOYMON011 g16508 BLASTN 498 1e-48 86 2025 16
701001311H1 SOYMON018 g16508 BLASTN 655 1e-48 85 2026 16
700734733H1 SOYMON010 g497899 BLASTN 682 1e-48 87 2027 16
701013070H1 SOYMON019 g497899 BLASTN 682 1e-48 87 2028 16
701120989H1 SOYMON037 g16508 BLASTN 683 1e-48 91 2029 16
LIB3049-025-Q1-E1-B4 LIB3049 g167961 BLASTN 687 1e-48 82 2030 16
700808455H1 SOYMON024 g16508 BLASTN 688 1e-48 87 2031 16
701101319H1 SOYMON028 g16508 BLASTN 688 1e-48 91 2032 16
701037087H1 SOYMON029 g609224 BLASTN 689 1e-48 85 2033 16
701205225H1 SOYMON035 g16508 BLASTN 689 1e-48 84 2034 16
701136807H1 SOYMON038 g609224 BLASTN 690 1e-48 85 2035 16
701118459H1 SOYMON037 g609224 BLASTN 690 1e-48 85 2036 16
700942825H1 SOYMON024 g16508 BLASTN 691 1e-48 85 2037 16
701139796H1 SOYMON038 g497899 BLASTN 692 1e-48 87 2038 16
700557040H1 SOYMON001 g609224 BLASTN 692 1e-48 85 2039 16
701015715H1 SOYMON038 g497899 BLASTN 692 1e-48 87 2040 16
700726424H1 SOYMON009 g497899 BLASTN 693 1e-48 87 2041 16
700790811H1 SOYMON011 g497899 BLASTN 693 1e-48 87 2042 16
LIB3040-038-Q1-E1-E5 LIB3040 g16508 BLASTN 709 1e-48 85 2043 16
700896626H1 SOYMON027 g429107 BLASTN 430 1e-47 84 2044 16
700562158H1 SOYMON002 g609224 BLASTN 518 1e-47 84 2045 16
700747095H1 SOYMON013 g16508 BLASTN 538 1e-47 87 2046 16
700726532H1 SOYMON009 g609224 BLASTN 622 1e-47 85 2047 16
701137686H1 SOYMON038 g497899 BLASTN 671 1e-47 85 2048 16
701009148H1 SOYMON019 g16508 BLASTN 673 1e-47 91 2049 16
701048279H1 SOYMON032 g16508 BLASTN 674 1e-47 86 2050 16
701120185H1 SOYMON037 g16508 BLASTN 674 1e-47 91 2051 16
701040355H1 SOYMON029 g497899 BLASTN 675 1e-47 86 2052 16
701130203H1 SOYMON037 g609224 BLASTN 678 1e-47 83 2053 16
701015794H1 SOYMON038 g16508 BLASTN 681 1e-47 91 2054 16
700732181H1 SOYMON010 g16508 BLASTN 681 1e-47 86 2055 16
LIB3050-018-Q1-E1-D10 LIB3050 g2305013 BLASTN 696 1e-47 81 2056 16
700899157H1 SOYMON027 g609224 BLASTN 486 1e-46 84 2057 16
701119905H1 SOYMON037 g497899 BLASTN 507 1e-46 86 2058 16
701062873H1 SOYMON033 g497899 BLASTN 509 1e-46 88 2059 16
701210886H1 SOYMON035 g16508 BLASTN 540 1e-46 84 2060 16
700893278H1 SOYMON024 g16508 BLASTN 542 1e-46 77 2061 16
701036970H1 SOYMON029 g16508 BLASTN 579 1e-46 88 2062 16
700901932H1 SOYMON027 g609224 BLASTN 640 1e-46 85 2063 16
701044214H1 SOYMON032 g429105 BLASTN 658 1e-46 85 2064 16
701056917H1 SOYMON033 g16508 BLASTN 661 1e-46 85 2065 16
700981560H1 SOYMON009 g497899 BLASTN 661 1e-46 87 2066 16
701213107H1 SOYMON035 g609224 BLASTN 661 1e-46 83 2067 16
700834235H1 SOYMON019 g497899 BLASTN 661 1e-46 87 2068 16
700749413H1 SOYMON013 g16508 BLASTN 662 1e-46 85 2069 16
700889148H1 SOYMON024 g16508 BLASTN 662 1e-46 85 2070 16
700745009H1 SOYMON013 g609224 BLASTN 662 1e-46 85 2071 16
701206618H1 SOYMON035 g16508 BLASTN 663 1e-46 91 2072 16
701131927H1 SOYMON038 g497899 BLASTN 666 1e-46 87 2073 16
701119069H1 SOYMON037 g497899 BLASTN 666 1e-46 87 2074 16
701106908H1 SOYMON036 g497899 BLASTN 666 1e-46 87 2075 16
701103063H1 SOYMON028 g497899 BLASTN 666 1e-46 87 2076 16
700686659H1 SOYMON008 g166873 BLASTN 667 1e-46 86 2077 16
701040620H1 SOYMON029 g16508 BLASTN 669 1e-46 86 2078 16
701012134H1 SOYMON019 g497899 BLASTN 669 1e-46 85 2079 16
700957508H1 SOYMON022 g1724103 BLASTN 669 1e-46 82 2080 16
701008548H1 SOYMON019 g497899 BLASTN 500 1e-45 87 2081 16
700978303H1 SOYMON009 g497899 BLASTN 506 1e-45 87 2082 16
701119990H1 SOYMON037 g497899 BLASTN 512 1e-45 87 2083 16
700646222H1 SOYMON012 g16508 BLASTN 526 1e-45 85 2084 16
701040631H1 SOYMON029 g609556 BLASTN 542 1e-45 87 2085 16
700894009H1 SOYMON024 g497899 BLASTN 646 1e-45 87 2086 16
700908702H1 SOYMON022 g609224 BLASTN 646 1e-45 85 2087 16
700838642H1 SOYMON020 g609224 BLASTN 646 1e-45 85 2088 16
700832089H1 SOYMON019 g609224 BLASTN 646 1e-45 85 2089 16
700978279H1 SOYMON009 g609224 BLASTN 647 1e-45 80 2090 16
700555058H1 SOYMON001 g16508 BLASTN 649 1e-45 84 2091 16
700906478H1 SOYMON022 g497899 BLASTN 651 1e-45 87 2092 16
700897237H1 SOYMON027 g16508 BLASTN 653 1e-45 86 2093 16
701125786H1 SOYMON037 g16508 BLASTN 654 1e-45 84 2094 16
700562836H1 SOYMON002 g16508 BLASTN 655 1e-45 84 2095 16
700665969H1 SOYMON005 g497899 BLASTN 656 1e-45 87 2096 16
701009838H1 SOYMON019 g497899 BLASTN 656 1e-45 87 2097 16
701211770H1 SOYMON035 g16508 BLASTN 657 1e-45 84 2098 16
700654550H1 SOYMON004 g16508 BLASTN 657 1e-45 83 2099 16
700567949H1 SOYMON002 g16508 BLASTN 657 1e-45 84 2100 16
701048175H1 SOYMON032 g16508 BLASTN 658 1e-45 91 2101 16
700901037H1 SOYMON027 g16508 BLASTN 658 1e-45 86 2102 16
700656917H1 SOYMON004 g1724103 BLASTN 407 1e-44 81 2103 16
700565684H1 SOYMON002 g16508 BLASTN 463 1e-44 86 2104 16
701002808H1 SOYMON019 g497899 BLASTN 503 1e-44 87 2105 16
701107115H1 SOYMON036 g497899 BLASTN 507 1e-44 87 2106 16
701122669H1 SOYMON037 g16508 BLASTN 636 1e-44 82 2107 16
700894409H1 SOYMON024 g497899 BLASTN 636 1e-44 87 2108 16
700848374H1 SOYMON021 g497899 BLASTN 636 1e-44 87 2109 16
700902420H1 SOYMON027 g497899 BLASTN 636 1e-44 87 2110 16
700747450H1 SOYMON013 g16508 BLASTN 638 1e-44 91 2111 16
701125869H1 SOYMON037 g16508 BLASTN 638 1e-44 91 2112 16
700731302H1 SOYMON010 g16508 BLASTN 639 1e-44 85 2113 16
701099068H1 SOYMON028 g2305013 BLASTN 640 1e-44 81 2114 16
700975556H1 SOYMON009 g450548 BLASTN 641 1e-44 84 2115 16
700889660H1 SOYMON024 g16508 BLASTN 642 1e-44 86 2116 16
700951744H1 SOYMON022 g16508 BLASTN 642 1e-44 86 2117 16
700750251H1 SOYMON013 g609224 BLASTN 643 1e-44 86 2118 16
700834611H1 SOYMON019 g16508 BLASTN 643 1e-44 91 2119 16
700565356H1 SOYMON002 g16508 BLASTN 643 1e-44 80 2120 16
700673803H1 SOYMON007 g16508 BLASTN 644 1e-44 84 2121 16
700958721H1 SOYMON022 g16508 BLASTN 645 1e-44 86 2122 16
700985717H1 SOYMON009 g16508 BLASTN 646 1e-44 82 2123 16
LIB3049-032-Q1-E1-A5 LIB3049 g16508 BLASTN 662 1e-44 76 2124 16
700733454H1 SOYMON010 g609224 BLASTN 432 1e-43 85 2125 16
700987656H1 SOYMON009 g497899 BLASTN 476 1e-43 87 2126 16
700755957H1 SOYMON014 g497899 BLASTN 483 1e-43 86 2127 16
700749331H1 SOYMON013 g2305013 BLASTN 521 1e-43 84 2128 16
700830984H1 SOYMON019 g497899 BLASTN 622 1e-43 86 2129 16
700906176H1 SOYMON022 g16508 BLASTN 623 1e-43 86 2130 16
701100070H2 SOYMON028 g16508 BLASTN 623 1e-43 91 2131 16
700954053H1 SOYMON022 g497899 BLASTN 624 1e-43 88 2132 16
700833949H1 SOYMON019 g497899 BLASTN 624 1e-43 88 2133 16
700976710H1 SOYMON009 g2305013 BLASTN 625 1e-43 79 2134 16
701117925H2 SOYMON037 g16508 BLASTN 625 1e-43 84 2135 16
700748825H1 SOYMON013 g16508 BLASTN 626 1e-43 84 2136 16
700899651H1 SOYMON027 g16508 BLASTN 626 1e-43 84 2137 16
700746739H1 SOYMON013 g497899 BLASTN 626 1e-43 88 2138 16
701009051H1 SOYMON019 g16508 BLASTN 628 1e-43 91 2139 16
700754423H1 SOYMON014 g16508 BLASTN 628 1e-43 91 2140 16
700760701H1 SOYMON015 g16508 BLASTN 628 1e-43 83 2141 16
701098124H1 SOYMON028 g2305013 BLASTN 628 1e-43 78 2142 16
700984979H1 SOYMON009 g16508 BLASTN 628 1e-43 91 2143 16
700565262H1 SOYMON002 g497899 BLASTN 629 1e-43 86 2144 16
700954979H1 SOYMON022 g497899 BLASTN 630 1e-43 87 2145 16
700834601H1 SOYMON019 g497899 BLASTN 630 1e-43 87 2146 16
700953156H1 SOYMON022 g609224 BLASTN 630 1e-43 85 2147 16
700865217H1 SOYMON016 g16508 BLASTN 631 1e-43 77 2148 16
700852947H1 SOYMON023 g16508 BLASTN 631 1e-43 84 2149 16
700943547H1 SOYMON024 g497899 BLASTN 631 1e-43 86 2150 16
701139277H1 SOYMON038 g609224 BLASTN 632 1e-43 82 2151 16
700900155H1 SOYMON027 g429105 BLASTN 264 1e-42 81 2152 16
700970581H1 SOYMON005 g497899 BLASTN 296 1e-42 86 2153 16
700654232H1 SOYMON003 g497899 BLASTN 348 1e-42 88 2154 16
700743608H1 SOYMON012 g497899 BLASTN 354 1e-42 86 2155 16
700986620H1 SOYMON009 g609224 BLASTN 360 1e-42 84 2156 16
700562590H1 SOYMON002 g16508 BLASTN 476 1e-42 83 2157 16
701119690H1 SOYMON037 g167961 BLASTN 506 1e-42 84 2158 16
700951730H1 SOYMON022 g497899 BLASTN 507 1e-42 87 2159 16
701004525H1 SOYMON019 g16508 BLASTN 516 1e-42 91 2160 16
700892637H1 SOYMON024 g16508 BLASTN 523 1e-42 81 2161 16
700560679H1 SOYMON001 g16508 BLASTN 526 1e-42 83 2162 16
700897888H1 SOYMON027 g16508 BLASTN 611 1e-42 86 2163 16
700547973H1 SOYMON001 g497899 BLASTN 611 1e-42 84 2164 16
701012166H1 SOYMON019 g497899 BLASTN 612 1e-42 87 2165 16
700747550H1 SOYMON013 g16508 BLASTN 613 1e-42 84 2166 16
700946136H1 SOYMON024 g16508 BLASTN 615 1e-42 85 2167 16
700665932H1 SOYMON005 g16508 BLASTN 616 1e-42 84 2168 16
701010969H1 SOYMON019 g16508 BLASTN 618 1e-42 84 2169 16
700962515H1 SOYMON022 g16508 BLASTN 618 1e-42 91 2170 16
700895685H1 SOYMON027 g429107 BLASTN 618 1e-42 77 2171 16
700752015H1 SOYMON014 g16508 BLASTN 620 1e-42 84 2172 16
701015704H1 SOYMON038 g16508 BLASTN 620 1e-42 85 2173 16
700966710H1 SOYMON028 g16508 BLASTN 620 1e-42 83 2174 16
701004268H1 SOYMON019 g16508 BLASTN 620 1e-42 86 2175 16
701138901H1 SOYMON038 g497899 BLASTN 285 1e-41 85 2176 16
700648891H1 SOYMON003 g16508 BLASTN 481 1e-41 84 2177 16
700741421H1 SOYMON012 g609224 BLASTN 511 1e-41 82 2178 16
700555653H1 SOYMON001 g166873 BLASTN 512 1e-41 85 2179 16
700958314H1 SOYMON022 g497899 BLASTN 599 1e-41 87 2180 16
700960055H1 SOYMON022 g497899 BLASTN 599 1e-41 87 2181 16
700901925H1 SOYMON027 g16508 BLASTN 599 1e-41 84 2182 16
700741987H1 SOYMON012 g497899 BLASTN 599 1e-41 87 2183 16
700662837H1 SOYMON005 g497899 BLASTN 599 1e-41 87 2184 16
701060927H1 SOYMON033 g497899 BLASTN 599 1e-41 87 2185 16
700562973H1 SOYMON002 g16508 BLASTN 601 1e-41 87 2186 16
700754586H1 SOYMON014 g16508 BLASTN 603 1e-41 84 2187 16
701132674H1 SOYMON038 g16508 BLASTN 604 1e-41 85 2188 16
701010012H2 SOYMON019 g609224 BLASTN 604 1e-41 84 2189 16
701123643H1 SOYMON037 g16508 BLASTN 604 1e-41 85 2190 16
700894196H1 SOYMON024 g16508 BLASTN 604 1e-41 85 2191 16
700899248H1 SOYMON027 g16508 BLASTN 604 1e-41 85 2192 16
701207680H1 SOYMON035 g16508 BLASTN 605 1e-41 84 2193 16
700899210H1 SOYMON027 g16508 BLASTN 605 1e-41 84 2194 16
700898762H1 SOYMON027 g16508 BLASTN 605 1e-41 87 2195 16
700983636H1 SOYMON009 g497899 BLASTN 606 1e-41 87 2196 16
700845511H1 SOYMON021 g16508 BLASTN 606 1e-41 89 2197 16
700563416H1 SOYMON002 g16508 BLASTN 606 1e-41 83 2198 16
701004041H1 SOYMON019 g16508 BLASTN 607 1e-41 84 2199 16
701097184H1 SOYMON028 g16508 BLASTN 607 1e-41 87 2200 16
700750691H1 SOYMON014 g609224 BLASTN 608 1e-41 84 2201 16
701208091H1 SOYMON035 g16508 BLASTN 608 1e-41 84
2202 16 700973888H1 SOYMON005 g609224 BLASTN 609 1e-41 85 2203 16
700831972H1 SOYMON019 g16508 BLASTN 609 1e-41 85 2204 16
701010090H2 SOYMON019 g16508 BLASTN 610 1e-41 87 2205 16
700890067H1 SOYMON024 g16508 BLASTN 610 1e-41 87 2206 16
700833227H1 SOYMON019 g16508 BLASTN 610 1e-41 85 2207 16
700563171H1 SOYMON002 g16508 BLASTN 370 1e-40 90 2208 16
701121457H1 SOYMON037 g609224 BLASTN 372 1e-40 85 2209 16
700566851H1 SOYMON002 g16508 BLASTN 402 1e-40 82 2210 16
700753221H1 SOYMON014 g609224 BLASTN 422 1e-40 84 2211 16
700749039H1 SOYMON013 g16508 BLASTN 475 1e-40 84 2212 16
700558940H1 SOYMON001 g16508 BLASTN 493 1e-40 80 2213 16
700757695H1 SOYMON015 g16508 BLASTN 526 1e-40 83 2214 16
701102262H1 SOYMON028 g2305013 BLASTN 587 1e-40 81 2215 16
700747484H1 SOYMON013 g16508 BLASTN 587 1e-40 85 2216 16
700891875H1 SOYMON024 g16508 BLASTN 587 1e-40 85 2217 16
700894439H1 SOYMON024 g497899 BLASTN 588 1e-40 88 2218 16
701062577H1 SOYMON033 g16508 BLASTN 588 1e-40 87 2219 16
700964366H1 SOYMON022 g16508 BLASTN 589 1e-40 85 2220 16
701210016H1 SOYMON035 g16508 BLASTN 589 1e-40 83 2221 16
700901573H1 SOYMON027 g16508 BLASTN 589 1e-40 85 2222 16
700750533H1 SOYMON014 g16508 BLASTN 589 1e-40 85 2223 16
700888820H1 SOYMON024 g497899 BLASTN 589 1e-40 87 2224 16
700891965H1 SOYMON024 g497899 BLASTN 589 1e-40 87 2225 16
700865001H1 SOYMON016 g16508 BLASTN 589 1e-40 85 2226 16
700654923H1 SOYMON004 g16508 BLASTN 590 1e-40 84 2227 16
700746169H1 SOYMON013 g16508 BLASTN 590 1e-40 84 2228 16
700745792H1 SOYMON013 g16508 BLASTN 591 1e-40 85 2229 16
700966953H1 SOYMON029 g16508 BLASTN 591 1e-40 85 2230 16
701141657H1 SOYMON038 g16508 BLASTN 592 1e-40 86 2231 16
700943078H1 SOYMON024 g16508 BLASTN 598 1e-40 85 2232 16
701137073H1 SOYMON038 g16508 BLASTN 598 1e-40 90 2233 16
700966730H1 SOYMON028 g16508 BLASTN 598 1e-40 84 2234 16
701137731H1 SOYMON038 g16508 BLASTN 371 1e-39 84 2235 16
701044050H1 SOYMON032 g497899 BLASTN 428 1e-39 89 2236 16
700993309H1 SOYMON011 g497899 BLASTN 434 1e-39 88 2237 16
700874483H1 SOYMON018 g497899 BLASTN 434 1e-39 87 2238 16
701014866H1 SOYMON019 g16508 BLASTN 454 1e-39 89 2239 16
700894603H1 SOYMON024 g16508 BLASTN 454 1e-39 85 2240 16
700958677H1 SOYMON022 g497899 BLASTN 455 1e-39 87 2241 16
700754001H1 SOYMON014 g609224 BLASTN 518 1e-39 85 2242 16
700946430H1 SOYMON024 g497899 BLASTN 578 1e-39 87 2243 16
700729892H1 SOYMON009 g497899 BLASTN 578 1e-39 87 2244 16
700753230H1 SOYMON014 g497899 BLASTN 578 1e-39 87 2245 16
700562584H1 SOYMON002 g16508 BLASTN 578 1e-39 82 2246 16
700842880H1 SOYMON020 g16508 BLASTN 580 1e-39 82 2247 16
700852561H1 SOYMON023 g16508 BLASTN 582 1e-39 85 2248 16
700941290H1 SOYMON024 g497899 BLASTN 583 1e-39 87 2249 16
700750361H1 SOYMON013 g497899 BLASTN 584 1e-39 80 2250 16
700841915H1 SOYMON020 g16508 BLASTN 585 1e-39 85 2251 16
700944462H1 SOYMON024 g16508 BLASTN 585 1e-39 85 2252 16
700548174H1 SOYMON002 g16508 BLASTN 342 1e-38 82 2253 16
700942324H1 SOYMON024 g497899 BLASTN 403 1e-38 87 2254 16
700788450H1 SOYMON011 g497899 BLASTN 416 1e-38 86 2255 16
701127954H1 SOYMON037 g16508 BLASTN 449 1e-38 76 2256 16
700742223H1 SOYMON012 g1724103 BLASTN 451 1e-38 77 2257 16
701102722H1 SOYMON028 g16508 BLASTN 466 1e-38 92 2258 16
701046957H1 SOYMON032 g16508 BLASTN 517 1e-38 85 2259 16
700836169H1 SOYMON019 g497899 BLASTN 517 1e-38 87 2260 16
700697967H1 SOYMON015 g16508 BLASTN 521 1e-38 84 2261 16
700678867H1 SOYMON007 g609224 BLASTN 566 1e-38 82 2262 16
700838753H1 SOYMON020 g16508 BLASTN 567 1e-38 85 2263 16
700726468H1 SOYMON009 g16508 BLASTN 570 1e-38 85 2264 16
701014631H1 SOYMON019 g497899 BLASTN 571 1e-38 87 2265 16
700728719H1 SOYMON009 g497899 BLASTN 573 1e-38 87 2266 16
700962582H1 SOYMON022 g497899 BLASTN 573 1e-38 87 2267 16
701123183H1 SOYMON037 g16508 BLASTN 573 1e-38 84 2268 16
700752347H1 SOYMON014 g16508 BLASTN 574 1e-38 87 2269 16
701103315H1 SOYMON028 g16508 BLASTN 394 1e-37 86 2270 16
700750955H1 SOYMON014 g497899 BLASTN 414 1e-37 88 2271 16
700867493H1 SOYMON016 g609224 BLASTN 433 1e-37 85 2272 16
701054760H1 SOYMON032 g16508 BLASTN 475 1e-37 85 2273 16
700900903H1 SOYMON027 g16508 BLASTN 501 1e-37 84 2274 16
701102294H1 SOYMON028 g16508 BLASTN 502 1e-37 87 2275 16
701124034H1 SOYMON037 g16508 BLASTN 512 1e-37 82 2276 16
700981476H1 SOYMON009 g16508 BLASTN 526 1e-37 83 2277 16
700903712H1 SOYMON022 g609224 BLASTN 552 1e-37 85 2278 16
700648751H1 SOYMON003 g1655577 BLASTN 554 1e-37 86 2279 16
700829930H1 SOYMON019 g609224 BLASTN 556 1e-37 84 2280 16
700758078H1 SOYMON015 g16508 BLASTN 556 1e-37 84 2281 16
700754248H1 SOYMON014 g16508 BLASTN 557 1e-37 81 2282 16
700834650H1 SOYMON019 g609224 BLASTN 557 1e-37 85 2283 16
700746279H1 SOYMON013 g16508 BLASTN 558 1e-37 82 2284 16
700869124H1 SOYMON016 g16508 BLASTN 559 1e-37 83 2285 16
701100219H1 SOYMON028 g2305013 BLASTN 560 1e-37 80 2286 16
700992589H1 SOYMON011 g497899 BLASTN 560 1e-37 80 2287 16
700865642H1 SOYMON016 g16508 BLASTN 562 1e-37 84 2288 16
701068835H1 SOYMON034 g429105 BLASTN 321 1e-36 86 2289 16
700746908H1 SOYMON013 g16508 BLASTN 373 1e-36 81 2290 16
700954719H1 SOYMON022 g16508 BLASTN 475 1e-36 86 2291 16
701042790H1 SOYMON029 g497899 BLASTN 489 1e-36 88 2292 16
700835259H1 SOYMON019 g497899 BLASTN 507 1e-36 88 2293 16
700565816H1 SOYMON002 g16508 BLASTN 519 1e-36 82 2294 16
700895811H1 SOYMON027 g609224 BLASTN 540 1e-36 83 2295 16
700975709H1 SOYMON009 g609224 BLASTN 541 1e-36 84 2296 16
700751645H1 SOYMON014 g16508 BLASTN 542 1e-36 85 2297 16
700567087H1 SOYMON002 g16508 BLASTN 542 1e-36 85 2298 16
700893027H1 SOYMON024 g609224 BLASTN 542 1e-36 85 2299 16
700891185H1 SOYMON024 g609224 BLASTN 543 1e-36 84 2300 16
700851376H1 SOYMON023 g16508 BLASTN 543 1e-36 83 2301 16
700898322H1 SOYMON027 g609224 BLASTN 547 1e-36 85 2302 16
700762881H1 SOYMON015 g16508 BLASTN 323 1e-35 85 2303 16
700960023H1 SOYMON022 g16508 BLASTN 370 1e-35 90 2304 16
701136486H1 SOYMON038 g497899 BLASTN 378 1e-35 85 2305 16
700987688H1 SOYMON009 g497899 BLASTN 387 1e-35 83 2306 16
700836508H1 SOYMON020 g16508 BLASTN 431 1e-35 85 2307 16
700990779H1 SOYMON011 g167961 BLASTN 471 1e-35 86 2308 16
700753019H1 SOYMON014 g497899 BLASTN 476 1e-35 83 2309 16
701138490H1 SOYMON038 g16508 BLASTN 481 1e-35 83 2310 16
700751771H1 SOYMON014 g609224 BLASTN 528 1e-35 84 2311 16
701048324H1 SOYMON032 g609224 BLASTN 529 1e-35 84 2312 16
700835739H1 SOYMON019 g16508 BLASTN 532 1e-35 85 2313 16
700986295H1 SOYMON009 g497899 BLASTN 533 1e-35 86 2314 16
701205693H1 SOYMON035 g16508 BLASTN 535 1e-35 84 2315 16
700865674H1 SOYMON016 g609224 BLASTN 536 1e-35 85 2316 16
700943777H1 SOYMON024 g16508 BLASTN 536 1e-35 84 2317 16
700968207H1 SOYMON035 g609224 BLASTN 536 1e-35 85 2318 16
700898370H1 SOYMON027 g609224 BLASTN 536 1e-35 85 2319 16
700987890H1 SOYMON009 g609224 BLASTN 536 1e-35 85 2320 16
700896016H1 SOYMON027 g609224 BLASTN 536 1e-35 85 2321 16
700554440H1 SOYMON001 g16508 BLASTN 283 1e-34 82 2322 16
700789855H2 SOYMON011 g16508 BLASTN 293 1e-34 91 2323 16
701101393H1 SOYMON028 g16508 BLASTN 311 1e-34 84 2324 16
700946335H1 SOYMON024 g16508 BLASTN 323 1e-34 86 2325 16
701207420H1 SOYMON035 g169664 BLASTN 350 1e-34 88 2326 16
700733034H1 SOYMON010 g16508 BLASTN 384 1e-34 81 2327 16
700790653H2 SOYMON011 g497899 BLASTN 399 1e-34 81 2328 16
700901808H1 SOYMON027 g497899 BLASTN 430 1e-34 81 2329 16
700760954H1 SOYMON015 g609224 BLASTN 514 1e-34 84 2330 16
700896206H1 SOYMON027 g16508 BLASTN 516 1e-34 85 2331 16
701039157H1 SOYMON029 g609224 BLASTN 520 1e-34 76 2332 16
700991056H1 SOYMON011 g166873 BLASTN 520 1e-34 80 2333 16
701097173H1 SOYMON028 g609224 BLASTN 521 1e-34 85 2334 16
700900989H1 SOYMON027 g497899 BLASTN 521 1e-34 86 2335 16
700895584H1 SOYMON027 g609224 BLASTN 521 1e-34 85 2336 16
700726668H1 SOYMON009 g16508 BLASTN 521 1e-34 85 2337 16
701011191H1 SOYMON019 g16508 BLASTN 521 1e-34 85 2338 16
701100827H1 SOYMON028 g16508 BLASTN 522 1e-34 84 2339 16
701156732H1 SOYMON031 g16508 BLASTN 522 1e-34 84 2340 16
700889752H1 SOYMON024 g609224 BLASTN 523 1e-34 84 2341 16
701105873H1 SOYMON036 g16508 BLASTN 524 1e-34 87 2342 16
700958346H1 SOYMON022 g497899 BLASTN 525 1e-34 84 2343 16
700962972H1 SOYMON022 g609224 BLASTN 525 1e-34 84 2344 16
701055918H1 SOYMON032 g16508 BLASTN 526 1e-34 85 2345 16
701123061H1 SOYMON037 g16508 BLASTN 526 1e-34 85 2346 16
701109796H1 SOYMON036 g166873 BLASTN 335 1e-33 86 2347 16
701207350H1 SOYMON035 g497899 BLASTN 349 1e-33 89 2348 16
700835692H1 SOYMON019 g16508 BLASTN 377 1e-33 81 2349 16
700729083H1 SOYMON009 g16508 BLASTN 448 1e-33 85 2350 16
LIB3040-049-Q1-E1-E8 LIB3040 g16508 BLASTN 464 1e-33 80 2351 16
700896567H1 SOYMON027 g497899 BLASTN 502 1e-33 81 2352 16
700668032H1 SOYMON006 g609224 BLASTN 504 1e-33 85 2353 16
700895062H1 SOYMON024 g609224 BLASTN 505 1e-33 83 2354 16
700961178H1 SOYMON022 g16508 BLASTN 506 1e-33 84 2355 16
701202590H1 SOYMON035 g497899 BLASTN 508 1e-33 86 2356 16
700754960H1 SOYMON014 g16508 BLASTN 512 1e-33 83 2357 16
700748584H1 SOYMON013 g16508 BLASTN 512 1e-33 84 2358 16
700791669H1 SOYMON011 g609224 BLASTN 512 1e-33 84 2359 16
701210405H1 SOYMON035 g16508 BLASTN 526 1e-33 83 2360 16
LIB3040-054-Q1-E1-D8 LIB3040 g16508 BLASTN 531 1e-33 80 2361 16
700791176H1 SOYMON011 g497899 BLASTN 311 1e-32 87 2362 16
LIB3049-029-Q1-E1-C5 LIB3049 g167961 BLASTN 368 1e-32 73 2363 16
700893458H1 SOYMON024 g497899 BLASTN 380 1e-32 87 2364 16
700829728H1 SOYMON019 g497899 BLASTN 490 1e-32 88 2365 16
700833137H1 SOYMON019 g609224 BLASTN 490 1e-32 85 2366 16
701204592H2 SOYMON035 g609224 BLASTN 493 1e-32 86 2367 16
700834821H1 SOYMON019 g609224 BLASTN 494 1e-32 78 2368 16
700757167H1 SOYMON015 g16508 BLASTN 494 1e-32 85 2369 16
701203730H2 SOYMON035 g16508 BLASTN 496 1e-32 82 2370 16
701044359H1 SOYMON032 g609224 BLASTN 497 1e-32 84 2371 16
701010095H2 SOYMON019 g16508 BLASTN 498 1e-32 86 2372 16
701100059H2 SOYMON028 g16508 BLASTN 499 1e-32 80 2373 16
700844256H1 SOYMON021 g16508 BLASTN 500 1e-32 80 2374 16
700968296H1 SOYMON035 g497899 BLASTN 501 1e-32 85 2375 16
701110404H1 SOYMON036 g16508 BLASTN 333 1e-31 84 2376 16
700869025H1 SOYMON016 g497899 BLASTN 340 1e-31 86 2377 16
700556245H1 SOYMON001 g609224 BLASTN 355 1e-31 84 2378 16
700566166H1 SOYMON002 g726031 BLASTN 402 1e-31 77 2379 16
700941481H1 SOYMON024 g497899 BLASTN 423 1e-31 79 2380 16
700896940H1 SOYMON027 g169664 BLASTN 477 1e-31 85 2381 16
701145429H1 SOYMON031 g609224 BLASTN 478 1e-31 85 2382 16
700946496H1 SOYMON024 g609224 BLASTN 488 1e-31 86 2383 16
700902149H1 SOYMON027 g1655577 BLASTN 488 1e-31 87 2384 16
700653072H1 SOYMON003 g497899 BLASTN 489 1e-31 88 2385 16
701000261H1 SOYMON018 g16508 BLASTN 495 1e-31 78 2386 16
701039358H1 SOYMON029 g609224 BLASTN 320 1e-30 85 2387 16
701156951H1 SOYMON031 g609224 BLASTN 467 1e-30 86 2388 16
701040880H1 SOYMON029 g497899 BLASTN 477 1e-30 89 2389 16
700979632H2 SOYMON009 g609224 BLASTN 490 1e-30 83 2390 16
700983678H1 SOYMON009 g16508 BLASTN 192 1e-29 86 2391 16
700893644H1 SOYMON024 g609224 BLASTN 320 1e-29 85 2392 16
700962437H1 SOYMON022 g609224 BLASTN 364 1e-29 83 2393 16
701150545H1 SOYMON031 g16508 BLASTN 457 1e-29 86 2394 16
700555403H1 SOYMON001 g16508 BLASTN 458 1e-29 90 2395 16
700794307H1 SOYMON017 g16508 BLASTN 460 1e-29 87 2396 16
701137971H1 SOYMON038 g16508 BLASTN 460 1e-29 84 2397 16
700893774H1 SOYMON024 g609224 BLASTN 461 1e-29 86 2398 16
700565272H1 SOYMON002 g609224 BLASTN 464 1e-29 74 2399 16
700667161H1 SOYMON006 g16508 BLASTN 468 1e-29 75 2400 16
700752952H1 SOYMON014 g609224 BLASTN 279 1e-28 78 2401 16
700901534H1 SOYMON027 g2305013 BLASTN 288 1e-28 85 2402 16
701006617H1 SOYMON019 g497899 BLASTN 358 1e-28 79 2403 16
700753459H1 SOYMON014 g609224 BLASTN 446 1e-28 81 2404 16
701157089H1 SOYMON031 g609224 BLASTN 448 1e-28 84 2405 16
700831271H1 SOYMON019 g16508 BLASTN 453 1e-28 87 2406 16
701207325H1 SOYMON035 g167961 BLASTN 251 1e-27 81 2407 16
700566121H1 SOYMON002 g16508 BLASTN 259 1e-27 75 2408 16
701137878H1 SOYMON038 g609224 BLASTN 280 1e-27 85 2409 16
700763957H1 SOYMON019 g497899 BLASTN 430 1e-27 88 2410 16
701015858H1 SOYMON038 g609224 BLASTN 431 1e-27 86 2411 16
700742849H1 SOYMON012 g497899 BLASTN 434 1e-27 88 2412 16
701102648H1 SOYMON028 g16508 BLASTN 436 1e-27 89 2413 16
700940955H1 SOYMON024 g609556 BLASTN 438 1e-27 85 2414 16
700889805H1 SOYMON024 g16508 BLASTN 441 1e-27 89 2415 16
701061930H1 SOYMON033 g16508 BLASTN 315 1e-26 77 2416 16
700901375H1 SOYMON027 g609224 BLASTN 323 1e-26 84 2417 16
700989056H1 SOYMON011 g609224 BLASTN 345 1e-26 82 2418 16
700665988H1 SOYMON005 g960356 BLASTN 420 1e-26 79 2419 16
700893352H1 SOYMON024 g497899 BLASTN 428 1e-26 87 2420 16
701122945H1 SOYMON037 g16508 BLASTN 269 1e-25 76 2421 16
700992929H1 SOYMON011 g16508 BLASTN 328 1e-25 78 2422 16
700557564H1 SOYMON001 g609224 BLASTN 413 1e-25 85 2423 16
701102620H1 SOYMON028 g609224 BLASTN 414 1e-25 82 2424 16
701062321H1 SOYMON033 g497899 BLASTN 267 1e-24 87 2425 16
701062258H1 SOYMON033 g450548 BLASTN 391 1e-24 83 2426 16
701145703H1 SOYMON031 g609224 BLASTN 411 1e-24 85 2427 16
701144943H1 SOYMON031 g450548 BLASTN 421 1e-24 85 2428 16
701134190H1 SOYMON038 g2305013 BLASTN 268 1e-23 87 2429 16
701132116H1 SOYMON038 g450548 BLASTN 385 1e-23 85 2430 16
701144522H1 SOYMON031 g450548 BLASTN 406 1e-23 85 2431 16
700738002H1 SOYMON012 g1655578 BLASTX 205 1e-21 84 2432 16
700756686H1 SOYMON014 g16508 BLASTN 235 1e-21 76 2433 16
700649058H1 SOYMON003 g16508 BLASTN 360 1e-21 85 2434 16
701152050H1 SOYMON031 g497899 BLASTN 233 1e-20 87 2435 16
701156991H1 SOYMON031 g497900 BLASTX 146 1e-19 98 2436 16
700761949H1 SOYMON015 g497900 BLASTX 173 1e-19 98 2437 16
700988163H1 SOYMON009 g609224 BLASTN 243 1e-19 81 2438 16
700666050H1 SOYMON005 g960357 BLASTX 183 1e-18 89 2439 16
700896896H1 SOYMON027 g497900 BLASTX 95 1e-17 82 2440 16
701100249H1 SOYMON028 g497900 BLASTX 104 1e-17 79 2441 16
701061719H1 SOYMON033 g609224 BLASTN 310 1e-17 67 2442 16
700674836H1 SOYMON007 g16508 BLASTN 331 1e-17 84 2443 16
700908822H1 SOYMON022 g169665 BLASTX 171 1e-16 100 2444 16
700898670H1 SOYMON027 g166872 BLASTX 172 1e-16 93 2445 16
700650967H1 SOYMON003 g16845 BLASTX 147 1e-15 100 2446 16
701132185H1 SOYMON038 g16845 BLASTX 151 1e-15 100 2447 16
700648929H1 SOYMON003 g1033190 BLASTX 156 1e-14 96 2448 16
700960809H1 SOYMON022 g16508 BLASTN 168 1e-14 88 2449 16
701101753H1 SOYMON028 g16845 BLASTX 141 1e-13 88 2450 16
700742010H1 SOYMON012 g609224 BLASTN 273 1e-13 88 2451 16
700735080H1 SOYMON010 g166874 BLASTX 121 1e-12 86 2452 16
700979242H1 SOYMON009 g609224 BLASTN 165 1e-12 81
2453 16 700740838H1 SOYMON012 g609224 BLASTN 253 1e-12 88 2454 16
700832383H1 SOYMON019 g16845 BLASTX 82 1e-11 71 2455 16 700830938H1
SOYMON019 g16845 BLASTX 92 1e-11 90 2456 16 700564686H1 SOYMON002
g1655578 BLASTX 95 1e-11 97 2457 16 700753178H1 SOYMON014 g16845
BLASTX 107 1e-10 78 2458 16 700563834H1 SOYMON002 g497900 BLASTX
124 1e-10 100 2459 16 700897739H1 SOYMON027 g16845 BLASTX 125 1e-10
86 2460 16 700725722H1 SOYMON009 g609225 BLASTX 125 1e-10 92 2461
16 701210357H1 SOYMON035 g497900 BLASTX 128 1e-10 100 2462 16
700833654H1 SOYMON019 g609224 BLASTN 243 1e-10 87 2463 16
701010334H1 SOYMON019 g609224 BLASTN 258 1e-10 86 2464 16
700567622H1 SOYMON002 g166874 BLASTX 93 1e-9 100 2465 16
700647933H1 SOYMON003 g609225 BLASTX 120 1e-9 81 2466 16
700742255H1 SOYMON012 g429107 BLASTN 160 1e-9 86 2467 16
700981506H1 SOYMON009 g609224 BLASTN 248 1e-9 88 2468 16
700962044H1 SOYMON022 g497900 BLASTX 113 1e-8 100 2469 16
701039604H1 SOYMON029 g497900 BLASTX 113 1e-8 100 2470 16
701009941H2 SOYMON019 g609225 BLASTX 114 1e-8 70 2471 16
700976956H1 SOYMON009 g609225 BLASTX 118 1e-8 96 2472 18138
701120722H1 SOYMON037 g169664 BLASTN 514 1e-33 92 2473 18138
700946044H1 SOYMON024 g169664 BLASTN 437 1e-26 90 2474 18138
700664443H1 SOYMON005 g17262 BLASTX 162 1e-16 88 2475 18138
700665162H1 SOYMON005 g17262 BLASTX 162 1e-16 88 2476 18138
701143667H1 SOYMON038 g16961 BLASTX 123 1e-11 95 2477 18138
701099150H1 SOYMON028 g16961 BLASTX 112 1e-9 90 2478 27686
700909141H1 SOYMON022 g726027 BLASTN 723 1e-51 84 2479 27686
701145458H1 SOYMON031 g726027 BLASTN 694 1e-49 86
ADENOSYLMETHIONINE DECARBOXYLASE (EC 4.1.1.50) 430 -700151703
700151703H1 SATMON007 g1532072 BLASTN 717 1e-50 91 431 -700165608
700165608H1 SATMON013 g1532072 BLASTN 605 1e-41 75 432 -700166868
700166868H1 SATMON013 g1532072 BLASTN 593 1e-40 90 433 -700242148
700242148H1 SATMON010 g1532048 BLASTX 142 1e-12 88 434 -700354923
700354923H1 SATMON024 g1532072 BLASTN 887 1e-74 89 435 -700422279
700422279H1 SATMONN01 g1532072 BLASTN 623 1e-61 88 436 -700455682
700455682H1 SATMON029 g1532047 BLASTN 681 1e-47 77 437 -700477645
700477645H1 SATMON025 g1532048 BLASTX 90 1e-12 71 438 -700550509
700550509H1 SATMON022 g1532047 BLASTN 399 1e-41 80 439 -700572502
700572502H1 SATMON030 g1532048 BLASTX 93 1e-20 70 440 -700618258
700618258H1 SATMON033 g1532072 BLASTN 368 1e-67 92 441 -701165456
701165456H1 SATMONN04 g1532072 BLASTN 436 1e-26 71 442 -L30622289
LIB3062-004-Q1-K1-E10 LIB3062 g1532047 BLASTN 406 1e-26 82 443
-L30623185 LIB3062-026-Q1-K1-E2 LIB3062 g1532072 BLASTN 338 1e-19
62 444 -L30625601 LIB3062-033-Q1-K1-A9 LIB3062 g1403043 BLASTN 758
1e-54 68 445 -L30661842 LIB3066-009-Q1-K1-E4 LIB3066 g1532072
BLASTN 398 1e-22 86 446 -L30681932 LIB3068-020-Q1-K1-C8 LIB3068
g1532072 BLASTN 205 1e-28 89 447 -L30687176 LIB3068-059-Q1-K1-C1
LIB3068 g1532072 BLASTN 216 1e-20 87 448 -L30692075
LIB3069-004-Q1-K1-G11 LIB3069 g1532072 BLASTN 619 1e-40 90 449
-L30783194 LIB3078-052-Q1-K1-E4 LIB3078 g1532072 BLASTN 447 1e-40
76 450 -L30792336 LIB3079-021-Q1-K1-E12 LIB3079 g1532072 BLASTN 217
1e-13 80 451 1471 700106762H1 SATMON010 g1532072 BLASTN 323 1e-59
84 452 1471 701163744H1 SATMONN04 g1532072 BLASTN 220 1e-41 86 453
1471 LIB3059-023-Q1-K1-A11 LIB3059 g1532072 BLASTN 340 1e-39 80 454
1471 700574530H1 SATMON030 g1532072 BLASTN 247 1e-34 82 455 1471
LIB83-003-Q1-E1-G1 LIB83 g1532072 BLASTN 239 1e-24 80 456 1471
701161147H1 SATMONN04 g1532072 BLASTN 239 1e-16 80 457 1471
700477336H1 SATMON025 g1532072 BLASTN 235 1e-10 91 458 16729
700169502H1 SATMON013 g1403043 BLASTN 456 1e-28 65 459 16729
700088201H1 SATMON011 g1532048 BLASTX 123 1e-22 60 460 16729
700165440H1 SATMON013 g1532048 BLASTX 162 1e-15 65 461 16866
700072905H1 SATMON007 g1532047 BLASTN 368 1e-19 71 462 16866
700350558H1 SATMON023 g1532047 BLASTN 301 1e-14 70 463 16866
700088370H1 SATMON011 g1532047 BLASTN 306 1e-14 70 464 2324
LIB3066-042-Q1-K1-H2 LIB3066 g1403043 BLASTN 1344 1e-103 79 465
2324 LIB3059-011-Q1-K1-B2 LIB3059 g1403043 BLASTN 1351 1e-103 78
466 2324 LIB3059-042-Q1-K1-B7 LIB3059 g1403043 BLASTN 1230 1e-100
80 467 2324 LIB3062-036-Q1-K1-F2 LIB3062 g1532072 BLASTN 1097 1e-91
80 468 2324 LIB3069-020-Q1-K1-A5 LIB3069 g1403043 BLASTN 925 1e-82
80 469 2324 LIB189-023-Q1-E1-H3 LIB189 g1532072 BLASTN 1085 1e-81
78 470 2324 700266088H1 SATMON017 g1532047 BLASTN 998 1e-74 80 471
2324 LIB3062-026-Q1-K1-E6 LIB3062 g1532047 BLASTN 982 1e-72 77 472
2324 700083335H1 SATMON011 g1403043 BLASTN 960 1e-71 79 473 2324
LIB189-015-Q1-E1-D3 LIB189 g1532047 BLASTN 970 1e-71 77 474 2324
LIB3079-013-Q1-K1-B3 LIB3079 g1403043 BLASTN 953 1e-70 81 475 2324
700262129H1 SATMON017 g1403043 BLASTN 936 1e-69 79 476 2324
700265667H1 SATMON017 g1403043 BLASTN 694 1e-66 81 477 2324
700807255H1 SATMON036 g1532072 BLASTN 850 1e-62 81 478 2324
700196129H1 SATMON014 g1403043 BLASTN 853 1e-62 83 479 2324
LIB3066-042-Q1-K1-H1 LIB3066 g1403043 BLASTN 842 1e-61 80 480 2324
700458321H1 SATMON029 g1403043 BLASTN 501 1e-58 80 481 2324
700197643H1 SATMON014 g1403043 BLASTN 461 1e-57 80 482 2324
LIB143-029-Q1-E1-H3 LIB143 g1532072 BLASTN 786 1e-56 78 483 2324
700197842H1 SATMON014 g1403043 BLASTN 688 1e-48 85 484 2324
700196807H1 SATMON014 g1532072 BLASTN 678 1e-47 75 485 2324
700263712H1 SATMON017 g1532072 BLASTN 403 1e-45 75 486 2324
LIB143-006-Q1-E1-F9 LIB143 g1403043 BLASTN 324 1e-42 83 487 2324
700267496H1 SATMON017 g1532047 BLASTN 594 1e-40 83 488 2324
700172551H1 SATMON013 g1532072 BLASTN 546 1e-36 74 489 2324
700211893H1 SATMON016 g1532047 BLASTN 542 1e-35 82 490 2324
700264065H1 SATMON017 g1532047 BLASTN 515 1e-34 83 491 2324
700465305H1 SATMON025 g1532073 BLASTX 148 1e-26 61 492 2324
700455052H1 SATMON029 g1403043 BLASTN 274 1e-26 79 493 2324
700473305H1 SATMON025 g1403043 BLASTN 446 1e-26 74 494 2324
700475851H1 SATMON025 g1532047 BLASTN 333 1e-18 78 495 2324
700263111H1 SATMON017 g1532073 BLASTX 165 1e-15 72 496 2324
700465705H1 SATMON025 g1403044 BLASTX 78 1e-8 76 497 3185
700264424H1 SATMON017 g1403043 BLASTN 469 1e-41 81 498 3185
LIB143-007-Q1-E1-H2 LIB143 g1403043 BLASTN 464 1e-36 80 499 3185
700262548H1 SATMON017 g1532047 BLASTN 368 1e-34 80 500 3185
700263845H1 SATMON017 g1403043 BLASTN 455 1e-34 81 501 3185
LIB3068-025-Q1-K1-G10 LIB3068 g1403043 BLASTN 288 1e-32 81 502 3185
700264569H1 SATMON017 g1403043 BLASTN 469 1e-32 82 503 3185
700243571H1 SATMON010 g1532047 BLASTN 461 1e-28 77 504 3185
700265453H1 SATMON017 g1532047 BLASTN 269 1e-27 84 505 3185
700267779H1 SATMON017 g1532047 BLASTN 257 1e-26 85 506 3185
700382373H1 SATMON024 g1532047 BLASTN 255 1e-24 86 507 3185
700258727H1 SATMON017 g1532047 BLASTN 255 1e-24 84 508 3185
LIB3069-018-Q1-K1-F5 LIB3069 g1532047 BLASTN 255 1e-21 77 509 3185
LIB3066-038-Q1-K1-C2 LIB3066 g1403043 BLASTN 242 1e-17 68 510 3185
700334654H1 SATMON019 g1532047 BLASTN 248 1e-17 90 511 3185
700262830H1 SATMON017 g1403043 BLASTN 276 1e-12 78 512 3185
700264727H1 SATMON017 g1532047 BLASTN 255 1e-10 92 513 3185
700238407H1 SATMON010 g1532047 BLASTN 255 1e-10 92 514 3185
700441952H1 SATMON026 g1532047 BLASTN 255 1e-10 92 515 3185
700268188H1 SATMON017 g1532047 BLASTN 255 1e-10 92 516 3185
700801627H1 SATMON036 g1532047 BLASTN 255 1e-10 92 517 3185
700261609H1 SATMON017 g1532047 BLASTN 255 1e-10 92 518 3185
700258510H1 SATMON017 g1532047 BLASTN 255 1e-10 92 519 3185
700258779H1 SATMON017 g1532047 BLASTN 255 1e-10 92 520 3185
700256838H1 SATMON017 g1532047 BLASTN 255 1e-10 92 521 3185
700263361H1 SATMON017 g1532047 BLASTN 255 1e-10 92 522 3185
700257102H1 SATMON017 g1532047 BLASTN 255 1e-10 92 523 3185
700239452H1 SATMON010 g1532047 BLASTN 245 1e-9 91 524 3185
700262045H1 SATMON017 g1532047 BLASTN 250 1e-9 90 525 8
LIB3066-027-Q1-K1-C6 LIB3066 g1532072 BLASTN 1414 1e-181 97 526 8
LIB3066-048-Q1-K1-C9 LIB3066 g1532072 BLASTN 1715 1e-173 98 527 8
LIB3066-007-Q1-K1-B4 LIB3066 g1532072 BLASTN 2146 1e-170 97 528 8
LIB3066-019-Q1-K1-B9 LIB3066 g1532072 BLASTN 1884 1e-168 98 529 8
LIB148-038-Q1-E1-A3 LIB148 g1532072 BLASTN 1585 1e-164 99 530 8
LIB3068-002-Q1-K1-H5 LIB3068 g1532072 BLASTN 2051 1e-162 98 531 8
LIB3067-035-Q1-K1-E9 LIB3067 g1532072 BLASTN 1594 1e-161 98 532 8
LIB189-020-Q1-E1-E8 LIB189 g1532072 BLASTN 1501 1e-160 98 533 8
LIB3078-057-Q1-K1-C12 LIB3078 g1532072 BLASTN 2026 1e-160 96 534 8
LIB3066-053-Q1-K1-A8 LIB3066 g1532072 BLASTN 1713 1e-159 93 535 8
LIB189-006-Q1-E1-C6 LIB189 g1532072 BLASTN 1676 1e-157 98 536 8
LIB3078-054-Q1-K1-E4 LIB3078 g1532072 BLASTN 1997 1e-157 95 537 8
LIB3069-028-Q1-K1-A11 LIB3069 g1532072 BLASTN 1980 1e-156 96 538 8
LIB3069-045-Q1-K1-H7 LIB3069 g1532072 BLASTN 1989 1e-156 98 539 8
LIB148-025-Q1-E1-F7 LIB148 g1532072 BLASTN 1954 1e-154 97 540 8
LIB189-014-Q1-E1-F11 LIB189 g1532072 BLASTN 1925 1e-151 94 541 8
LIB3060-014-Q1-K1-C5 LIB3060 g1532072 BLASTN 1780 1e-150 96 542 8
LIB148-057-Q1-E1-C2 LIB148 g1532072 BLASTN 1000 1e-149 98 543 8
LIB148-015-Q1-E1-F2 LIB148 g1532072 BLASTN 1886 1e-148 94 544 8
LIB3066-045-Q1-K1-A2 LIB3066 g1532072 BLASTN 914 1e-146 91 545 8
LIB148-064-Q1-E1-A3 LIB148 g1532072 BLASTN 1660 1e-146 93 546 8
LIB3061-047-Q1-K1-G1 LIB3061 g1532072 BLASTN 1867 1e-146 92 547 8
LIB148-040-Q1-E1-F1 LIB148 g1532072 BLASTN 1706 1e-145 94 548 8
LIB3069-012-Q1-K1-B6 LIB3069 g1532072 BLASTN 1630 1e-144 87 549 8
LIB3068-009-Q1-K1-A8 LIB3068 g1532072 BLASTN 1802 1e-141 97 550 8
LIB148-004-Q1-E1-D2 LIB148 g1532072 BLASTN 818 1e-139 91 551 8
LIB3068-002-Q1-K1-A1 LIB3068 g1532072 BLASTN 944 1e-137 95 552 8
LIB3067-035-Q1-K1-H9 LIB3067 g1532072 BLASTN 1701 1e-137 98 553 8
LIB3068-033-Q1-K1-G12 LIB3068 g1532072 BLASTN 1093 1e-130 89 554 8
700572229H1 SATMON030 g1532072 BLASTN 1135 1e-128 99 555 8
LIB3078-052-Q1-K1-E1 LIB3078 g1532072 BLASTN 1235 1e-127 85 556 8
700572579H1 SATMON030 g1532072 BLASTN 1625 1e-126 98 557 8
LIB3066-025-Q1-K1-F2 LIB3066 g1532072 BLASTN 1625 1e-126 100 558 8
LIB3068-048-Q1-K1-F9 LIB3068 g1532072 BLASTN 1538 1e-125 98 559 8
700098413H1 SATMON009 g1532072 BLASTN 1595 1e-124 100 560 8
700573235H1 SATMON030 g1532072 BLASTN 1513 1e-123 98 561 8
700090946H1 SATMON011 g1532072 BLASTN 1585 1e-123 100 562 8
700092465H1 SATMON008 g1532072 BLASTN 1541 1e-122 98 563 8
700074625H1 SATMON007 g1532072 BLASTN 1570 1e-122 100 564 8
LIB3059-007-Q1-K1-C10 LIB3059 g1532072 BLASTN 1401 1e-119 94 565 8
700072828H1 SATMON007 g1532072 BLASTN 1540 1e-119 100 566 8
700619106H1 SATMON034 g1532072 BLASTN 833 1e-118 97 567 8
700074853H1 SATMON007 g1532072 BLASTN 1515 1e-117 100 568 8
700201293H1 SATMON003 g1532072 BLASTN 1517 1e-117 97 569 8
700075896H1 SATMON007 g1532072 BLASTN 1006 1e-115 99 570 8
LIB3059-052-Q1-K1-A1 LIB3059 g1532072 BLASTN 1241 1e-115 92 571 8
700091576H1 SATMON011 g1532072 BLASTN 943 1e-114 98 572 8
LIB3078-015-Q1-K1-D7 LIB3078 g1532072 BLASTN 1218 1e-114 87 573 8
700074733H1 SATMON007 g1532072 BLASTN 1475 1e-114 100 574 8
700381421H1 SATMON023 g1532072 BLASTN 1475 1e-114 100 575 8
700085594H1 SATMON011 g1532072 BLASTN 1477 1e-114 99 576 8
700095883H1 SATMON008 g1532072 BLASTN 1477 1e-114 99 577 8
700338237H1 SATMON020 g1532072 BLASTN 1478 1e-114 99 578 8
700549813H1 SATMON022 g1532072 BLASTN 1481 1e-114 99 579 8
700097935H1 SATMON009 g1532072 BLASTN 1484 1e-114 98 580 8
700572978H1 SATMON030 g1532072 BLASTN 865 1e-113 96 581 8
700196464H1 SATMON014 g1532072 BLASTN 1044 1e-113 92 582 8
700381412H1 SATMON023 g1532072 BLASTN 1168 1e-113 98 583 8
700027839H1 SATMON003 g1532072 BLASTN 1472 1e-113 99 584 8
700623344H1 SATMON034 g1532072 BLASTN 1085 1e-112 94 585 8
700025858H1 SATMON003 g1532072 BLASTN 1455 1e-112 100 586 8
LIB3059-017-Q1-K1-H5 LIB3059 g1532072 BLASTN 1459 1e-112 97 587 8
700475019H1 SATMON025 g1532072 BLASTN 1390 1e-111 100 588 8
700338357H1 SATMON020 g1532072 BLASTN 1440 1e-111 100 589 8
700256796H1 SATMON017 g1532072 BLASTN 1400 1e-110 100 590 8
700071692H1 SATMON007 g1532072 BLASTN 1430 1e-110 98 591 8
700339073H1 SATMON020 g1532072 BLASTN 1357 1e-109 95 592 8
700106916H1 SATMON010 g1532072 BLASTN 1415 1e-109 93 593 8
700468234H1 SATMON025 g1532072 BLASTN 1420 1e-109 100 594 8
700214482H1 SATMON016 g1532072 BLASTN 1425 1e-109 100 595 8
700466437H1 SATMON025 g1532072 BLASTN 1403 1e-108 98 596 8
700205576H1 SATMON003 g1532072 BLASTN 1404 1e-108 98 597 8
700043455H1 SATMON004 g1532072 BLASTN 1405 1e-108 100 598 8
700348439H1 SATMON023 g1532072 BLASTN 1407 1e-108 99 599 8
700093131H1 SATMON008 g1532072 BLASTN 755 1e-107 100 600 8
700571851H1 SATMON030 g1532072 BLASTN 1302 1e-107 99 601 8
700088142H1 SATMON011 g1532072 BLASTN 1392 1e-107 99 602 8
700085934H1 SATMON011 g1532072 BLASTN 1397 1e-107 98 603 8
700028465H1 SATMON003 g1532072 BLASTN 1258 1e-106 98 604 8
700236818H1 SATMON010 g1532072 BLASTN 1380 1e-106 100 605 8
700583691H1 SATMON031 g1532072 BLASTN 1382 1e-106 99 606 8
700095585H1 SATMON008 g1532072 BLASTN 1383 1e-106 94 607 8
700105140H1 SATMON010 g1532072 BLASTN 1375 1e-105 100 608 8
700338486H1 SATMON020 g1532072 BLASTN 893 1e-104 98 609 8
700214178H1 SATMON016 g1532072 BLASTN 1361 1e-104 99 610 8
700090613H1 SATMON011 g1532072 BLASTN 699 1e-103 99 611 8
700576189H1 SATMON030 g1532072 BLASTN 1254 1e-103 93 612 8
700475734H1 SATMON025 g1532072 BLASTN 1256 1e-103 99 613 8
700028218H1 SATMON003 g1532072 BLASTN 1275 1e-103 100 614 8
700088644H1 SATMON011 g1532072 BLASTN 1351 1e-103 98 615 8
700378768H1 SATMON020 g1532072 BLASTN 1067 1e-102 98 616 8
LIB3067-035-Q1-K1-H10 LIB3067 g1532072 BLASTN 1233 1e-102 95 617 8
700043466H1 SATMON004 g1532072 BLASTN 1331 1e-102 97 618 8
LIB148-057-Q1-E1-C3 LIB148 g1532072 BLASTN 1283 1e-101 92 619 8
700217574H1 SATMON016 g1532072 BLASTN 1320 1e-101 100 620 8
700042332H1 SATMON004 g1532072 BLASTN 1321 1e-101 97 621 8
700440926H1 SATMON026 g1532072 BLASTN 1323 1e-101 99 622 8
700552284H1 SATMON022 g1532072 BLASTN 1329 1e-101 97 623 8
700216613H1 SATMON016 g1532072 BLASTN 689 1e-100 97 624 8
LIB3067-054-Q1-K1-F6 LIB3067 g1532072 BLASTN 1030 1e-100 94 625 8
LIB3060-038-Q1-K1-B9 LIB3060 g1532072 BLASTN 1108 1e-100 92 626 8
700218755H1 SATMON011 g1532072 BLASTN 1116 1e-100 99 627 8
700338042H1 SATMON020 g1532072 BLASTN 1270 1e-100 99 628 8
700268009H1 SATMON017 g1532072 BLASTN 1307 1e-100 92 629 8
700578491H1 SATMON031 g1532072 BLASTN 1309 1e-100 97 630 8
700030037H1 SATMON003 g1532072 BLASTN 1310 1e-100 97 631 8
700221406H1 SATMON011 g1532072 BLASTN 1315 1e-100 100 632 8
700157357H1 SATMON012 g1532072 BLASTN 1315 1e-100 98 633 8
700476926H1 SATMON025 g1532072 BLASTN 680 1e-99 97 634 8
700475068H1 SATMON025 g1532072 BLASTN 792 1e-99 98 635 8
700469741H1 SATMON025 g1532072 BLASTN 831 1e-99 99 636 8
700082377H1 SATMON011 g1532072 BLASTN 1081 1e-99 99 637 8
700217143H1 SATMON016 g1532072 BLASTN 1208 1e-99 98 638 8
700339433H1 SATMON020 g1532072 BLASTN 1295 1e-99 100 639 8
700196627H1 SATMON014 g1532072 BLASTN 1295 1e-99 100 640 8
700259354H1 SATMON017 g1532072 BLASTN 1305 1e-99 92 641 8
700610842H1 SATMON022 g1532072 BLASTN 843 1e-98 97 642 8
700218059H1 SATMON016 g1532072 BLASTN 1036 1e-98 99 643 8
700421727H1 SATMONN01 g1532072 BLASTN 1172 1e-98 97 644 8
700158660H1 SATMON012 g1532072 BLASTN 1285 1e-98 100 645 8
700806856H1 SATMON036 g1532072 BLASTN 1194 1e-97 97 646 8
700583512H1 SATMON031 g1532072 BLASTN 1219 1e-97 97 647 8
700025502H1 SATMON004 g1532072 BLASTN 1276 1e-97 99 648 8
700159562H1 SATMON012 g1532072 BLASTN 1277 1e-97 99 649 8
700223731H1 SATMON011 g1532072 BLASTN 1277 1e-97 99 650 8
700156494H1 SATMON012 g1532072 BLASTN 1280 1e-97 100 651 8
700160533H1 SATMON012 g1532072 BLASTN 1281 1e-97 97 652 8
LIB3066-055-Q1-K1-D8 LIB3066 g1532072 BLASTN 672 1e-96 99
653 8 700405152H1 SATMON028 g1532072 BLASTN 828 1e-96 98 654 8
LIB148-040-Q1-E1-A8 LIB148 g1532072 BLASTN 1268 1e-96 83 655 8
700438437H1 SATMON026 g1532072 BLASTN 1246 1e-95 99 656 8
700267245H1 SATMON017 g1532072 BLASTN 1248 1e-95 93 657 8
700156129H2 SATMON007 g1532072 BLASTN 1250 1e-95 100 658 8
700203246H1 SATMON003 g1532072 BLASTN 1250 1e-95 100 659 8
700168339H1 SATMON013 g1532072 BLASTN 915 1e-94 98 660 8
LIB3067-036-Q1-K1-F9 LIB3067 g1532072 BLASTN 1110 1e-94 95 661 8
700193769H1 SATMON014 g1532072 BLASTN 1240 1e-94 100 662 8
700094014H1 SATMON008 g1532072 BLASTN 1241 1e-94 91 663 8
700020311H1 SATMON001 g1532072 BLASTN 1228 1e-93 99 664 8
700195083H1 SATMON014 g1532072 BLASTN 1229 1e-93 98 665 8
700193901H1 SATMON014 g1532072 BLASTN 1229 1e-93 98 666 8
700194639H1 SATMON014 g1532072 BLASTN 1231 1e-93 99 667 8
700350117H1 SATMON023 g1532072 BLASTN 1143 1e-92 99 668 8
700239867H1 SATMON010 g1532072 BLASTN 1210 1e-92 98 669 8
LIB3069-028-Q1-K1-D1 LIB3069 g1532072 BLASTN 1217 1e-92 94 670 8
700355756H1 SATMON024 g1532072 BLASTN 618 1e-91 96 671 8
700265492H1 SATMON017 g1532072 BLASTN 620 1e-91 94 672 8
700579021H1 SATMON031 g1532072 BLASTN 656 1e-91 96 673 8
LIB3079-015-Q1-K1-B11 LIB3079 g1532072 BLASTN 1008 1e-91 83 674 8
700572888H2 SATMON030 g1532072 BLASTN 1159 1e-91 99 675 8
701183990H1 SATMONN06 g1532072 BLASTN 1198 1e-91 93 676 8
700085757H1 SATMON011 g1532072 BLASTN 1200 1e-91 100 677 8
700162756H1 SATMON013 g1532072 BLASTN 1208 1e-91 99 678 8
700193076H1 SATMON014 g1532072 BLASTN 1208 1e-91 99 679 8
700224378H1 SATMON011 g1532072 BLASTN 696 1e-90 99 680 8
700218773H1 SATMON011 g1532072 BLASTN 1187 1e-90 93 681 8
700159338H1 SATMON012 g1532072 BLASTN 1193 1e-90 97 682 8
700169217H1 SATMON013 g1532072 BLASTN 1196 1e-90 99 683 8
700551548H1 SATMON022 g1532072 BLASTN 839 1e-89 96 684 8
700197059H1 SATMON014 g1532072 BLASTN 1025 1e-89 95 685 8
700469834H1 SATMON025 g1532072 BLASTN 650 1e-88 97 686 8
700569778H1 SATMON030 g1532072 BLASTN 682 1e-88 91 687 8
700194845H1 SATMON014 g1532072 BLASTN 1171 1e-88 99 688 8
700445861H1 SATMON027 g1532072 BLASTN 536 1e-87 99 689 8
700100536H1 SATMON009 g1532072 BLASTN 666 1e-87 94 690 8
701163801H1 SATMONN04 g1532072 BLASTN 805 1e-87 94 691 8
LIB3060-035-Q1-K1-E3 LIB3060 g1532072 BLASTN 922 1e-87 95 692 8
700170872H1 SATMON013 g1532072 BLASTN 943 1e-87 97 693 8
700241270H1 SATMON010 g1532072 BLASTN 1066 1e-86 96 694 8
700457981H1 SATMON029 g1532072 BLASTN 1140 1e-86 92 695 8
700158339H1 SATMON012 g1532072 BLASTN 1142 1e-86 99 696 8
700019442H1 SATMON001 g1532072 BLASTN 1145 1e-86 100 697 8
700149885H1 SATMON007 g1532072 BLASTN 1133 1e-85 99 698 8
700018255H1 SATMON001 g1532072 BLASTN 1135 1e-85 100 699 8
700244026H1 SATMON010 g1532072 BLASTN 1044 1e-84 89 700 8
LIB3060-027-Q1-K1-B2 LIB3060 g1532072 BLASTN 1117 1e-84 99 701 8
700170960H1 SATMON013 g1532072 BLASTN 696 1e-83 99 702 8
700267487H1 SATMON017 g1532072 BLASTN 1061 1e-83 92 703 8
700152754H1 SATMON007 g1532072 BLASTN 1107 1e-83 99 704 8
700378260H1 SATMON019 g1532072 BLASTN 1112 1e-83 92 705 8
700455089H1 SATMON029 g1532072 BLASTN 583 1e-82 97 706 8
700167322H1 SATMON013 g1532072 BLASTN 1080 1e-81 98 707 8
LIB3060-035-Q1-K1-H1 LIB3060 g1532072 BLASTN 782 1e-80 94 708 8
700204067H1 SATMON003 g1532072 BLASTN 1011 1e-80 98 709 8
700049271H1 SATMON003 g1532072 BLASTN 627 1e-79 93 710 8
700442065H1 SATMON026 g1532072 BLASTN 1043 1e-78 91 711 8
700244175H1 SATMON010 g1532072 BLASTN 1044 1e-78 90 712 8
700045296H1 SATMON004 g1532072 BLASTN 1041 1e-77 93 713 8
700578391H1 SATMON031 g1532072 BLASTN 624 1e-76 88 714 8
700346136H1 SATMON021 g1532072 BLASTN 751 1e-76 90 715 8
700457852H1 SATMON029 g1532072 BLASTN 777 1e-76 92 716 8
700570111H1 SATMON030 g1532072 BLASTN 814 1e-75 94 717 8
LIB3066-030-Q1-K1-A12 LIB3066 g1532072 BLASTN 922 1e-75 94 718 8
700149657H1 SATMON007 g1532072 BLASTN 1012 1e-75 92 719 8
LIB3060-043-Q1-K1-B4 LIB3060 g1532072 BLASTN 1013 1e-75 97 720 8
700156752H1 SATMON012 g1532072 BLASTN 1013 1e-75 98 721 8
700454114H1 SATMON029 g1532072 BLASTN 455 1e-74 93 722 8
LIB143-053-Q1-E1-E8 LIB143 g1532072 BLASTN 672 1e-74 98 723 8
700029323H1 SATMON003 g1532072 BLASTN 996 1e-74 99 724 8
700156381H1 SATMON007 g1532072 BLASTN 1002 1e-74 97 725 8
700159603H2 SATMON012 g1532072 BLASTN 1003 1e-74 94 726 8
LIB3060-035-Q1-K1-E5 LIB3060 g1532072 BLASTN 603 1e-72 94 727 8
LIB3069-025-Q1-K1-E12 LIB3069 g1532072 BLASTN 734 1e-72 90 728 8
700166680H1 SATMON013 g1532072 BLASTN 971 1e-72 99 729 8
700171123H1 SATMON013 g1532072 BLASTN 973 1e-72 94 730 8
LIB148-042-Q1-E1-G10 LIB148 g1532072 BLASTN 975 1e-72 100 731 8
700612951H1 SATMON033 g1532072 BLASTN 401 1e-71 96 732 8
700265213H1 SATMON017 g1532072 BLASTN 626 1e-71 92 733 8
LIB189-030-Q1-E1-D11 LIB189 g1532072 BLASTN 740 1e-70 92 734 8
700212877H1 SATMON016 g1532072 BLASTN 918 1e-70 93 735 8
700161545H1 SATMON012 g1532072 BLASTN 621 1e-69 99 736 8
700021259H1 SATMON001 g1532072 BLASTN 940 1e-69 93 737 8
LIB3079-015-Q1-K1-C7 LIB3079 g1532072 BLASTN 508 1e-68 96 738 8
LIB3066-055-Q1-K1-F11 LIB3066 g1532072 BLASTN 832 1e-68 85 739 8
700166771H1 SATMON013 g1532072 BLASTN 922 1e-68 88 740 8
LIB3066-003-Q1-K1-E6 LIB3066 g1532072 BLASTN 927 1e-68 86 741 8
700208131H1 SATMON016 g1532047 BLASTN 747 1e-66 85 742 8
700020688H1 SATMON001 g1532072 BLASTN 905 1e-66 90 743 8
700206405H1 SATMON003 g1532072 BLASTN 905 1e-66 100 744 8
LIB3069-042-Q1-K1-C5 LIB3069 g1532072 BLASTN 891 1e-65 99 745 8
700353835H1 SATMON024 g1532072 BLASTN 896 1e-65 96 746 8
700471533H1 SATMON025 g1532072 BLASTN 496 1e-63 99 747 8
700165425H1 SATMON013 g1532072 BLASTN 850 1e-62 100 748 8
LIB3069-054-Q1-K1-C4 LIB3069 g1532072 BLASTN 613 1e-60 87 749 8
700160896H1 SATMON012 g1532072 BLASTN 746 1e-60 95 750 8
LIB3069-042-Q1-K1-A11 LIB3069 g1532072 BLASTN 840 1e-60 98 751 8
700466625H1 SATMON025 g1532072 BLASTN 635 1e-59 86 752 8
700571770H1 SATMON030 g1532072 BLASTN 820 1e-59 84 753 8
LIB143-024-Q1-E1-D2 LIB143 g1532072 BLASTN 813 1e-58 95 754 8
700453677H1 SATMON028 g1532072 BLASTN 508 1e-57 92 755 8
700193979H1 SATMON014 g1532072 BLASTN 790 1e-57 100 756 8
700467828H1 SATMON025 g1532072 BLASTN 779 1e-56 96 757 8
700150072H1 SATMON007 g1532072 BLASTN 776 1e-55 99 758 8
700802869H1 SATMON036 g1532072 BLASTN 702 1e-54 95 759 8
LIB148-051-Q1-E1-B12 LIB148 g1532072 BLASTN 421 1e-53 92 760 8
700075035H1 SATMON007 g1532072 BLASTN 506 1e-53 93 761 8
700471283H1 SATMON025 g1532072 BLASTN 742 1e-53 91 762 8
700166625H1 SATMON013 g1532072 BLASTN 748 1e-53 99 763 8
700019894H1 SATMON001 g1532072 BLASTN 643 1e-51 89 764 8
700204863H1 SATMON003 g1532072 BLASTN 721 1e-51 99 765 8
700431071H1 SATMONN01 g1532072 BLASTN 366 1e-50 94 766 8
700171582H1 SATMON013 g1532072 BLASTN 492 1e-50 99 767 8
700450579H1 SATMON028 g1532072 BLASTN 338 1e-49 97 768 8
700084149H1 SATMON011 g1532072 BLASTN 672 1e-47 98 769 8
700095070H1 SATMON008 g1532072 BLASTN 662 1e-46 98 770 8
700570827H1 SATMON030 g1532072 BLASTN 344 1e-44 83 771 8
LIB3061-057-Q1-K1-G12 LIB3061 g1532072 BLASTN 378 1e-43 75 772 8
700194918H1 SATMON014 g1532072 BLASTN 626 1e-43 89 773 8
700378894H1 SATMON020 g1532072 BLASTN 495 1e-42 97 774 8
700468029H1 SATMON025 g1532072 BLASTN 616 1e-42 98 775 8
LIB143-012-Q1-E1-H8 LIB143 g1532072 BLASTN 637 1e-42 98 776 8
LIB3060-037-Q1-K1-H4 LIB3060 g1532072 BLASTN 638 1e-42 86 777 8
700433327H1 SATMONN01 g1532072 BLASTN 365 1e-41 86 778 8
700623611H1 SATMON034 g1532072 BLASTN 316 1e-39 95 779 8
700166123H1 SATMON013 g1532072 BLASTN 585 1e-39 100 780 8
700616273H1 SATMON033 g1532072 BLASTN 534 1e-38 98 781 8
700464702H1 SATMON025 g1532072 BLASTN 566 1e-38 99 782 8
700338809H1 SATMON020 g1532072 BLASTN 550 1e-37 100 783 8
700092622H1 SATMON008 g1532072 BLASTN 561 1e-37 99 784 8
LIB3060-043-Q1-K1-A10 LIB3060 g1532072 BLASTN 352 1e-36 93 785 8
700265611H1 SATMON017 g1532072 BLASTN 548 1e-36 95 786 8
700100319H1 SATMON009 g1532072 BLASTN 552 1e-36 98 787 8
700092696H1 SATMON008 g1532072 BLASTN 552 1e-36 98 788 8
700076750H1 SATMON007 g1532072 BLASTN 526 1e-35 99 789 8
700082896H1 SATMON011 g1532072 BLASTN 526 1e-35 99 790 8
700075411H1 SATMON007 g1532072 BLASTN 319 1e-34 91 791 8
701178051H1 SATMONN05 g1532072 BLASTN 342 1e-34 94 792 8
700266358H1 SATMON017 g1532072 BLASTN 520 1e-34 95 793 8
700453981H1 SATMON029 g1532072 BLASTN 505 1e-33 96 794 8
700584238H1 SATMON031 g1532072 BLASTN 509 1e-33 91 795 8
700103871H1 SATMON010 g1532072 BLASTN 491 1e-32 99 796 8
700264460H1 SATMON017 g1532072 BLASTN 495 1e-32 95 797 8
700205122H1 SATMON003 g1532072 BLASTN 501 1e-32 99 798 8
700165768H1 SATMON013 g1532072 BLASTN 420 1e-31 98 799 8
700077179H1 SATMON007 g1532072 BLASTN 471 1e-30 98 800 8
700476654H1 SATMON025 g1532072 BLASTN 476 1e-30 98 801 8
700027374H1 SATMON003 g1532072 BLASTN 476 1e-30 98 802 8
700266994H1 SATMON017 g1532072 BLASTN 476 1e-30 98 803 8
700088193H1 SATMON011 g1532072 BLASTN 488 1e-30 97 804 8
700214511H1 SATMON016 g1532072 BLASTN 298 1e-29 93 805 8
700257186H1 SATMON017 g1532072 BLASTN 465 1e-29 95 806 8
700335814H1 SATMON019 g1532072 BLASTN 469 1e-29 93 807 8
700236166H1 SATMON010 g1532072 BLASTN 450 1e-28 95 808 8
700468243H1 SATMON025 g1532072 BLASTN 456 1e-28 98 809 8
700096327H1 SATMON008 g1532072 BLASTN 451 1e-27 98 810 8
700471139H1 SATMON025 g1532072 BLASTN 451 1e-27 98 811 8
700050420H1 SATMON003 g1532072 BLASTN 365 1e-26 92 812 8
700264846H1 SATMON017 g1532072 BLASTN 430 1e-25 94 813 8
700266803H1 SATMON017 g1532072 BLASTN 293 1e-24 75 814 8
700086134H1 SATMON011 g1532072 BLASTN 422 1e-24 97 815 8
700162001H1 SATMON012 g1532072 BLASTN 279 1e-23 96 816 8
700044825H1 SATMON004 g1532072 BLASTN 411 1e-23 98 817 8
700074679H1 SATMON007 g1532072 BLASTN 411 1e-23 98 818 8
700267694H1 SATMON017 g1532072 BLASTN 320 1e-22 92 819 8
700267990H1 SATMON017 g1532072 BLASTN 361 1e-22 99 820 8
700456715H1 SATMON029 g1532072 BLASTN 293 1e-21 97 821 8
700454806H1 SATMON029 g1532072 BLASTN 356 1e-21 98 822 8
700466812H1 SATMON025 g1532072 BLASTN 380 1e-21 95 823 8
700046476H1 SATMON004 g1532072 BLASTN 381 1e-21 97 824 8
700207160H1 SATMON017 g1532072 BLASTN 393 1e-21 96 825 8
700383037H1 SATMON024 g1532072 BLASTN 280 1e-20 98 826 8
700549660H1 SATMON022 g1532072 BLASTN 376 1e-20 97 827 8
LIB3066-004-Q1-K1-G12 LIB3066 g1532072 BLASTN 379 1e-20 91 828 8
700801422H1 SATMON036 g1532072 BLASTN 321 1e-19 97 829 8
700045319H1 SATMON004 g1532072 BLASTN 336 1e-17 99 830 8
700046047H1 SATMON004 g1532072 BLASTN 336 1e-17 99 831 8
700264328H1 SATMON017 g1532072 BLASTN 339 1e-17 95 832 8
700083485H1 SATMON011 g1532072 BLASTN 341 1e-17 99 833 8
700461268H1 SATMON033 g1532072 BLASTN 342 1e-17 95 834 8
700053271H1 SATMON008 g1532072 BLASTN 311 1e-15 98 835 8
700215275H1 SATMON016 g1532072 BLASTN 321 1e-15 98 836 8
700623133H1 SATMON034 g1532072 BLASTN 275 1e-14 75 837 8
700454949H1 SATMON029 g1532072 BLASTN 280 1e-14 99 838 8
700440763H1 SATMON026 g1532072 BLASTN 307 1e-14 95 839 8
LIB3068-022-Q1-K1-C4 LIB3068 g1532072 BLASTN 150 1e-12 94 840 8
700798849H1 SATMON036 g1532072 BLASTN 281 1e-12 98 841 8
700333057H1 SATMON019 g1532072 BLASTN 184 1e-11 91 842 8
700265786H1 SATMON017 g1532072 BLASTN 200 1e-11 90 843 8
700193030H1 SATMON014 g1532072 BLASTN 158 1e-9 94 844 8 700262964H1
SATMON017 g1532047 BLASTN 171 1e-9 81 845 8 LIB3069-023-Q1-K1-B2
LIB3069 g1403043 BLASTN 195 1e-9 84 846 8 700474096H1 SATMON025
g1532072 BLASTN 199 1e-9 97 847 8 700028326H1 SATMON003 g1532072
BLASTN 151 1e-8 96 848 8011 700440327H1 SATMON026 g1532047 BLASTN
292 1e-14 85 849 851 LIB3059-052-Q1-K1-E6 LIB3059 g1403043 BLASTN
1033 1e-85 75 850 851 700090958H1 SATMON011 g1532072 BLASTN 1044
1e-78 80 851 851 700224605H1 SATMON011 g1532072 BLASTN 850 1e-62 80
852 851 700551562H1 SATMON022 g1403043 BLASTN 374 1e-55 79 853 851
700153939H1 SATMON007 g1403043 BLASTN 772 1e-55 80 854 851
700469231H1 SATMON025 g1532072 BLASTN 560 1e-52 76 855 851
700349854H1 SATMON023 g1403043 BLASTN 669 1e-46 81 856 851
701169324H1 SATMONN05 g1403043 BLASTN 669 1e-46 78 857 851
700088319H1 SATMON011 g1532072 BLASTN 492 1e-30 78 2480 -700998660
700998660H1 SOYMON018 g1531764 BLASTN 249 1e-29 90 2481 -GM927
LIB3028-005-Q1-B1-B4 LIB3028 g1421750 BLASTN 261 1e-10 80 2482
13379 700842514H1 SOYMON020 g1915980 BLASTN 575 1e-41 73 2483 13379
701042786H1 SOYMON029 g1421750 BLASTN 377 1e-20 80 2484 15556
701101584H1 SOYMON028 g1421750 BLASTN 526 1e-35 69 2485 15556
701099749H1 SOYMON028 g1155239 BLASTN 453 1e-27 72 2486 16
LIB3051-018-Q1-E1-A7 LIB3051 g1421750 BLASTN 1146 1e-93 81 2487 16
LIB3056-005-Q1-N1-E10 LIB3056 g1421750 BLASTN 944 1e-88 80 2488 16
LIB3051-101-Q1-K1-C7 LIB3051 g1421750 BLASTN 1135 1e-85 80 2489 16
LIB3056-001-Q1-B1-H12 LIB3056 g1421750 BLASTN 752 1e-82 81 2490 16
700661334H1 SOYMON005 g1421750 BLASTN 1092 1e-82 80 2491 16
LIB3056-012-Q1-N1-B12 LIB3056 g1421750 BLASTN 1085 1e-81 80 2492 16
LIB3055-011-Q1-N1-G2 LIB3055 g1421750 BLASTN 958 1e-78 79 2493 16
700663981H1 SOYMON005 g1421750 BLASTN 996 1e-74 82 2494 16
701109737H1 SOYMON036 g1421750 BLASTN 955 1e-70 83 2495 16
701130190H1 SOYMON037 g1421750 BLASTN 935 1e-69 81 2496 16
701003301H1 SOYMON019 g1421750 BLASTN 412 1e-68 82 2497 16
700894137H1 SOYMON024 g1421750 BLASTN 923 1e-68 84 2498 16
700980096H1 SOYMON009 g1421750 BLASTN 923 1e-68 81 2499 16
700942625H1 SOYMON024 g1421750 BLASTN 911 1e-67 80 2500 16
700829695H1 SOYMON019 g1421750 BLASTN 918 1e-67 84 2501 16
700662572H1 SOYMON005 g1421750 BLASTN 920 1e-67 84 2502 16
701127647H1 SOYMON037 g1421750 BLASTN 902 1e-66 84 2503 16
701014550H1 SOYMON019 g1421750 BLASTN 556 1e-65 81 2504 16
LIB3051-043-Q1-K1-G3 LIB3051 g1421750 BLASTN 698 1e-65 82 2505 16
700730914H1 SOYMON009 g1421750 BLASTN 897 1e-65 84 2506 16
701052455H1 SOYMON032 g1421750 BLASTN 876 1e-64 81 2507 16
LIB3051-074-Q1-K1-A7 LIB3051 g1421750 BLASTN 876 1e-64 75 2508 16
700874839H1 SOYMON018 g1421750 BLASTN 862 1e-63 82 2509 16
701005943H1 SOYMON019 g1421750 BLASTN 865 1e-63 82 2510 16
700663357H1 SOYMON005 g1421750 BLASTN 869 1e-63 82 2511 16
701042053H1 SOYMON029 g1421750 BLASTN 872 1e-63 81 2512 16
700971988H1 SOYMON005 g1421750 BLASTN 508 1e-61 82 2513 16
701010728H1 SOYMON019 g1421750 BLASTN 657 1e-61 81 2514 16
700987206H1 SOYMON009 g1421750 BLASTN 845 1e-61 83 2515 16
701126773H1 SOYMON037 g1421750 BLASTN 847 1e-61 79 2516 16
700764714H1 SOYMON023 g1421750 BLASTN 830 1e-60 81 2517 16
700974444H1 SOYMON005 g1421750 BLASTN 830 1e-60 81 2518 16
701118620H1 SOYMON037 g1421750 BLASTN 832 1e-60 75 2519 16
700729926H1 SOYMON009 g1421750 BLASTN 834 1e-60 79 2520 16
700945142H1 SOYMON024 g1421750 BLASTN 470 1e-59 84 2521 16
700867912H1 SOYMON016 g1421750 BLASTN 820 1e-59 82 2522 16
700746546H1 SOYMON013 g1421750 BLASTN 824 1e-59 78 2523 16
700873331H1 SOYMON018 g1421750 BLASTN 769 1e-58 82 2524 16
700738212H1 SOYMON012 g1421750 BLASTN 780 1e-56 83 2525 16
700830454H1 SOYMON019 g1421750 BLASTN 789 1e-56 81
2526 16 700726129H1 SOYMON009 g1421750 BLASTN 789 1e-56 81 2527 16
700996774H1 SOYMON018 g1421750 BLASTN 486 1e-55 81 2528 16
700746427H1 SOYMON013 g1421750 BLASTN 766 1e-55 77 2529 16
701123911H1 SOYMON037 g1421750 BLASTN 769 1e-55 76 2530 16
700745730H1 SOYMON013 g1421750 BLASTN 775 1e-55 76 2531 16
700900940H1 SOYMON027 g1421750 BLASTN 760 1e-54 78 2532 16
700846419H1 SOYMON021 g1421750 BLASTN 762 1e-54 78 2533 16
701203427H1 SOYMON035 g1421750 BLASTN 491 1e-53 84 2534 16
LIB3051-074-Q1-K1-F5 LIB3051 g1421750 BLASTN 650 1e-53 75 2535 16
700747646H1 SOYMON013 g1421750 BLASTN 747 1e-53 77 2536 16
701049823H1 SOYMON032 g1421750 BLASTN 751 1e-53 77 2537 16
701049188H1 SOYMON032 g1421750 BLASTN 731 1e-52 75 2538 16
700875156H1 SOYMON018 g1421750 BLASTN 737 1e-52 78 2539 16
700864540H1 SOYMON016 g1421750 BLASTN 741 1e-52 81 2540 16
700682329H2 SOYMON008 g1421750 BLASTN 719 1e-51 76 2541 16
700846215H1 SOYMON021 g1421750 BLASTN 575 1e-50 82 2542 16
701101327H1 SOYMON028 g1421750 BLASTN 592 1e-50 77 2543 16
LIB3055-011-Q1-N1-E7 LIB3055 g1421750 BLASTN 680 1e-50 80 2544 16
700848992H1 SOYMON021 g1421750 BLASTN 708 1e-50 76 2545 16
700966820H1 SOYMON028 g1421750 BLASTN 710 1e-50 75 2546 16
701051785H1 SOYMON032 g1421750 BLASTN 717 1e-50 75 2547 16
700681079H1 SOYMON008 g1917012 BLASTN 290 1e-49 77 2548 16
701138880H1 SOYMON038 g1421750 BLASTN 610 1e-49 78 2549 16
700872967H1 SOYMON018 g1421750 BLASTN 704 1e-49 75 2550 16
701119960H1 SOYMON037 g1421750 BLASTN 684 1e-48 81 2551 16
700873306H1 SOYMON018 g1421750 BLASTN 693 1e-48 83 2552 16
700751484H1 SOYMON014 g1421750 BLASTN 635 1e-47 82 2553 16
700958865H1 SOYMON022 g1421750 BLASTN 486 1e-46 82 2554 16
700872833H1 SOYMON018 g1421750 BLASTN 659 1e-46 84 2555 16
700871673H1 SOYMON018 g1421750 BLASTN 659 1e-46 84 2556 16
700872801H1 SOYMON018 g1421750 BLASTN 659 1e-46 84 2557 16
700847572H1 SOYMON021 g1421750 BLASTN 660 1e-46 75 2558 16
700728046H1 SOYMON009 g1421750 BLASTN 661 1e-46 84 2559 16
700894721H1 SOYMON024 g1421750 BLASTN 550 1e-43 84 2560 16
700730946H1 SOYMON009 g1421750 BLASTN 307 1e-42 77 2561 16
700727943H1 SOYMON009 g1421750 BLASTN 378 1e-42 79 2562 16
701101238H1 SOYMON028 g1421750 BLASTN 456 1e-42 92 2563 16
700758519H1 SOYMON015 g1421750 BLASTN 358 1e-41 77 2564 16
700740343H1 SOYMON012 g1531764 BLASTN 607 1e-41 72 2565 16
700848076H1 SOYMON021 g1531764 BLASTN 469 1e-40 89 2566 16
LIB3051-080-Q1-K1-H7 LIB3051 g1531764 BLASTN 472 1e-38 90 2567 16
700955916H1 SOYMON022 g1531764 BLASTN 427 1e-37 90 2568 16
700873876H1 SOYMON018 g1421750 BLASTN 465 1e-37 80 2569 16
700746271H1 SOYMON013 g1421750 BLASTN 515 1e-37 80 2570 16
700662887H1 SOYMON005 g1421750 BLASTN 506 1e-35 82 2571 16
700983591H1 SOYMON009 g1421750 BLASTN 537 1e-35 82 2572 16
700752343H1 SOYMON014 g1421750 BLASTN 488 1e-34 73 2573 16
700901973H1 SOYMON027 g1421750 BLASTN 490 1e-34 82 2574 16
LIB3053-013-Q1-N1-B11 LIB3053 g1421750 BLASTN 522 1e-34 75 2575 16
700988481H1 SOYMON009 g1421750 BLASTN 506 1e-33 78 2576 16
LIB3051-078-Q1-K1-C12 LIB3051 g1421750 BLASTN 302 1e-32 75 2577 16
700985640H1 SOYMON009 g1421750 BLASTN 449 1e-30 86 2578 16
701127038H1 SOYMON037 g1421750 BLASTN 467 1e-30 67 2579 16
700898677H1 SOYMON027 g1531764 BLASTN 474 1e-30 89 2580 16
700742260H1 SOYMON012 g1421750 BLASTN 476 1e-30 70 2581 16
701103203H1 SOYMON028 g1421750 BLASTN 287 1e-27 81 2582 16
700897144H1 SOYMON027 g1421750 BLASTN 264 1e-26 75 2583 16
700832039H1 SOYMON019 g1421750 BLASTN 390 1e-26 82 2584 16
700743311H1 SOYMON012 g1421750 BLASTN 429 1e-26 87 2585 16
LIB3040-044-Q1-E1-B8 LIB3040 g1421750 BLASTN 282 1e-25 92 2586 16
LIB3051-024-Q1-K1-C4 LIB3051 g2394382 BLASTX 110 1e-24 95 2587 16
701011765H1 SOYMON019 g1421750 BLASTN 383 1e-21 76 2588 16
700893428H1 SOYMON024 g1421752 BLASTX 106 1e-20 82 2589 16
700996204H1 SOYMON018 g1421750 BLASTN 288 1e-20 75 2590 16
LIB3049-028-Q1-E1-C6 LIB3049 g1421750 BLASTN 230 1e-19 90 2591 16
701103628H1 SOYMON028 g1421750 BLASTN 277 1e-18 73 2592 16
700972636H1 SOYMON005 g1421750 BLASTN 286 1e-16 73 2593 16
700875417H1 SOYMON018 g1421750 BLASTN 209 1e-15 75 2594 16
701207702H1 SOYMON035 g1421752 BLASTX 153 1e-14 83 2595 16
701054556H1 SOYMON032 g1421750 BLASTN 230 1e-14 75 2596 16
701046407H1 SOYMON032 g1421750 BLASTN 230 1e-14 75 2597 16
701117634H1 SOYMON037 g1421750 BLASTN 230 1e-14 74 2598 16
701127714H1 SOYMON037 g1421750 BLASTN 230 1e-14 75 2599 16
700663239H1 SOYMON005 g1421750 BLASTN 210 1e-13 74 2600 16
700738510H1 SOYMON012 g1421750 BLASTN 212 1e-13 72 2601 16
700943559H1 SOYMON024 g1421750 BLASTN 215 1e-13 71 2602 16
700666278H1 SOYMON005 g1421750 BLASTN 290 1e-13 93 2603 16
700663144H1 SOYMON005 g1421750 BLASTN 204 1e-12 74 2604 16
700684147H1 SOYMON008 g1421750 BLASTN 278 1e-12 89 2605 16
700953076H1 SOYMON022 g1421750 BLASTN 285 1e-12 92 2606 16
701099931H1 SOYMON028 g1421750 BLASTN 202 1e-11 72 2607 16
701058910H1 SOYMON033 g1421750 BLASTN 216 1e-11 74 2608 16
700754949H1 SOYMON014 g1421750 BLASTN 266 1e-11 89 2609 16
700562967H1 SOYMON002 g1421750 BLASTN 266 1e-11 82 2610 16
701012790H1 SOYMON019 g1421750 BLASTN 251 1e-10 89 2611 16
701101739H1 SOYMON028 g1421750 BLASTN 260 1e-10 82 2612 16
701118211H1 SOYMON037 g1490554 BLASTX 97 1e-9 47 2613 16
700844394H1 SOYMON021 g1917013 BLASTX 116 1e-9 85 2614 16
701048560H1 SOYMON032 g1421750 BLASTN 198 1e-9 79 2615 16
701062373H1 SOYMON033 g1421750 BLASTN 203 1e-9 71 2616 16
701120496H1 SOYMON037 g1421750 BLASTN 230 1e-9 82 2617 16
701049680H1 SOYMON032 g1421750 BLASTN 177 1e-8 70 2618 16
700748760H1 SOYMON013 g1421750 BLASTN 230 1e-8 91 2619 16
700889482H1 SOYMON024 g1421750 BLASTN 235 1e-8 91 2620 16
700725013H1 SOYMON009 g1421750 BLASTN 239 1e-8 85 2621 16048
LIB3028-004-Q1-B1-E2 LIB3028 g1421750 BLASTN 879 1e-64 68 2622
16048 700761667H1 SOYMON015 g1421750 BLASTN 528 1e-35 71 2623 16048
700958003H1 SOYMON022 g1421750 BLASTN 428 1e-26 78 ASPARTATE KINASE
(EC 2.7.2.4) 858 -700018870 700018870H1 SATMON001 g500850 BLASTN
1095 1e-82 100 859 -700085903 700085903H1 SATMON011 g500852 BLASTN
1606 1e-124 99 860 -700086169 700086169H1 SATMON011 g500852 BLASTN
559 1e-94 92 861 -700096679 700096679H1 SATMON008 g500850 BLASTN
1131 1e-85 99 862 -700096794 700096794H1 SATMON008 g500850 BLASTN
1515 1e-117 100 863 -700106390 700106390H1 SATMON010 g2243115
BLASTN 544 1e-51 72 864 -700168286 700168286H1 SATMON013 g2243115
BLASTN 519 1e-34 72 865 -700171363 700171363H1 SATMON013 g500850
BLASTN 1085 1e-81 94 866 -700194781 700194781H1 SATMON014 g2257742
BLASTN 635 1e-44 79 867 -700213839 700213839H1 SATMON016 g500850
BLASTN 664 1e-75 94 868 -700219756 700219756H1 SATMON011 g500850
BLASTN 1359 1e-104 99 869 -700258808 700258808H1 SATMON017 g500850
BLASTN 630 1e-85 96 870 -700263439 700263439H1 SATMON017 g2243115
BLASTN 448 1e-43 76 871 -700266615 700266615H1 SATMON017 g2243116
BLASTX 179 1e-17 73 872 -700342655 700342655H1 SATMON021 g2257743
BLASTX 213 1e-22 82 873 -700343285 700343285H1 SATMON021 g2257742
BLASTN 437 1e-25 66 874 -700467533 700467533H1 SATMON025 g2243115
BLASTN 439 1e-26 79 875 -700548678 700548678H1 SATMON022 g147979
BLASTX 147 1e-13 62 876 -700613618 700613618H1 SATMON033 g500850
BLASTN 965 1e-81 98 877 -L30691987 LIB3069-018-Q1-K1-A3 LIB3069
g2243115 BLASTN 266 1e-10 57 878 12201 700457103H1 SATMON029
g2257742 BLASTN 789 1e-56 76 879 12201 700457111H1 SATMON029
g2243115 BLASTN 513 1e-33 76 880 12931 700380864H1 SATMON023
g500852 BLASTN 1450 1e-111 95 881 12931 700105610H1 SATMON010
g500852 BLASTN 1046 1e-105 95 882 12931 700380848H1 SATMON023
g500852 BLASTN 1193 1e-96 95 883 12931 700205392H1 SATMON003
g500852 BLASTN 1065 1e-94 93 884 12931 700552314H1 SATMON022
g500852 BLASTN 908 1e-86 90 885 12931 700551915H1 SATMON022 g500852
BLASTN 1090 1e-81 90 886 16037 700344509H1 SATMON021 g500852 BLASTN
1184 1e-89 92 887 16037 700345170H1 SATMON021 g500852 BLASTN 600
1e-58 85 888 16157 700212607H1 SATMON016 g2243115 BLASTN 914 1e-67
76 889 16157 700094809H1 SATMON008 g2243116 BLASTX 144 1e-12 84 890
19231 700091761H1 SATMON011 g500850 BLASTN 1358 1e-104 98 891 19231
700612568H1 SATMON033 g500850 BLASTN 1137 1e-102 99 892 22303
700553291H1 SATMON022 g2243115 BLASTN 644 1e-44 70 893 22303
700553382H1 SATMON022 g2243115 BLASTN 418 1e-39 70 894 28000
LIB143-061-Q1-E1-C7 LIB143 g2243115 BLASTN 1132 1e-85 75 895 28000
700474110H1 SATMON025 g2243115 BLASTN 509 1e-33 75 896 30401
700620948H1 SATMON034 g500852 BLASTN 327 1e-30 88 897 32907
LIB143-038-Q1-E1-B11 LIB143 g500850 BLASTN 1864 1e-146 96 898 32907
700096779H1 SATMON008 g500850 BLASTN 1490 1e-115 97 899 5616
700346488H1 SATMON021 g2243115 BLASTN 664 1e-46 71 900 5616
700196138H1 SATMON014 g2243115 BLASTN 630 1e-43 74 2624 -700556108
700556108H1 SOYMON001 g2243115 BLASTN 700 1e-49 74 2625 -700663367
700663367H1 SOYMON005 g2243115 BLASTN 737 1e-52 77 2626 -700733301
700733301H1 SOYMON010 g2243115 BLASTN 751 1e-53 78 2627 -700747979
700747979H1 SOYMON013 g2257742 BLASTN 449 1e-27 70 2628 -700832664
700832664H1 SOYMON019 g167547 BLASTN 322 1e-44 78 2629 -700843925
700843925H1 SOYMON021 g167547 BLASTN 616 1e-42 71 2630 -700888516
700888516H1 SOYMON024 g464225 BLASTX 193 1e-19 78 2631 -700892002
700892002H1 SOYMON024 g2243115 BLASTN 363 1e-21 79 2632 -700959057
700959057H1 SOYMON022 g2257742 BLASTN 497 1e-32 72 2633 -700971891
700971891H1 SOYMON005 g167547 BLASTN 699 1e-49 74 2634 -700984812
700984812H1 SOYMON009 g2257742 BLASTN 801 1e-57 77 2635 -701069254
701069254H1 SOYMON034 g2243115 BLASTN 260 1e-23 75 2636 -701120341
701120341H1 SOYMON037 g2243115 BLASTN 567 1e-38 77 2637 -GM35173
LIB3051-037-Q1-K1-B9 LIB3051 g2970554 BLASTN 193 1e-11 83 2638
15020 700557507H1 SOYMON001 g167547 BLASTN 914 1e-67 79 2639 15020
700666142H1 SOYMON005 g1107460 BLASTN 767 1e-55 77 2640 18237
700797368H1 SOYMON017 g2257742 BLASTN 819 1e-59 81 2641 18237
700797360H1 SOYMON017 g2257742 BLASTN 809 1e-58 84 2642 19332
LIB3056-004-Q1-N1-D5 LIB3056 g2243115 BLASTN 1118 1e-84 74 2643
19332 700786255H2 SOYMON011 g2257742 BLASTN 626 1e-43 72 2644 19332
700684751H1 SOYMON008 g2257742 BLASTN 583 1e-39 73 2645 21954
701100440H1 SOYMON028 g167547 BLASTN 835 1e-60 78 2646 21954
701059173H1 SOYMON033 g167547 BLASTN 719 1e-51 75 2647 26336
701003103H1 SOYMON019 g2243115 BLASTN 877 1e-64 79 2648 26336
700976874H1 SOYMON009 g2243115 BLASTN 880 1e-64 79
ASPARTATE-SEMIALDEHYDE DEHYDROGENASE (EC 1.2.1.11) 901 -700439614
700439614H1 SATMON026 g2314350 BLASTX 107 1e-13 52 902 -701183695
701183695H1 SATMONN06 g289910 BLASTX 150 1e-13 69 903 -L1487398
LIB148-064-Q1-E1-D10 LIB148 g1749466 BLASTX 178 1e-33 55 904
-L30622830 LIB3062-028-Q1-K1-A8 LIB3062 g1085109 BLASTX 108 1e-40
48 2649 -700756763 700756763H1 SOYMON014 g1359593 BLASTX 71 1e-8 52
2650 -700830054 700830054H1 SOYMON019 g142828 BLASTX 87 1e-10 41
2651 -701105617 701105617H1 SOYMON036 g142828 BLASTX 215 1e-22 57
2652 -GM8539 LIB3039-047-Q1-E1-C6 LIB3039 g142828 BLASTX 188 1e-35
50 2653 30187 LIB3049-001-Q1-E1-F2 LIB3049 g1359593 BLASTX 136
1e-26 54 2654 30187 700556105H1 SOYMON001 g1359593 BLASTX 132 1e-11
56 O-SUCCINYLHOMOSERINE (THIOL)-LYASE (EC 4.2.99.9) 905 -700049526
700049526H1 SATMON003 g2198852 BLASTN 251 1e-9 76 906 -700086788
700086788H1 SATMON011 g2198850 BLASTN 631 1e-61 88 907 -700460589
700460589H1 SATMON030 g2198850 BLASTN 251 1e-32 83 908 -700577561
700577561H1 SATMON031 g2198852 BLASTN 207 1e-15 82 909 -700579840
700579840H1 SATMON031 g2198850 BLASTN 216 1e-12 94 910 -700616013
700616013H1 SATMON033 g2198852 BLASTN 278 1e-39 81 911 -L1892785
LIB189-011-Q1-E1-A10 LIB189 g2198852 BLASTN 311 1e-14 82 912
-L30594402 LIB3059-042-Q1-K1-E11 LIB3059 g2198852 BLASTN 489 1e-40
84 913 -L30604293 LIB3060-028-Q1-K1-E7 LIB3060 g2198852 BLASTN 380
1e-20 83 914 -L30693289 LIB3069-016-Q1-K1-G10 LIB3069 g2198852
BLASTN 197 1e-10 79 915 10571 700224881H1 SATMON011 g2198852 BLASTN
319 1e-15 75 916 10801 700429049H1 SATMONN01 g2198852 BLASTN 210
1e-16 82 917 10801 700167813H1 SATMON013 g2198852 BLASTN 200 1e-15
82 918 10801 700074027H1 SATMON007 g2198852 BLASTN 178 1e-11 80 919
16379 LIB3061-035-Q1-K1-B6 LIB3061 g2198850 BLASTN 2184 1e-173 99
920 16379 700259309H1 SATMON017 g2198850 BLASTN 1259 1e-108 92 921
16379 700051696H1 SATMON003 g2198850 BLASTN 1392 1e-107 96 922
16379 700239553H1 SATMON010 g2198850 BLASTN 1265 1e-96 96 923 16379
700042846H1 SATMON004 g2198850 BLASTN 1208 1e-91 95 924 16379
700092741H1 SATMON008 g2198850 BLASTN 1042 1e-89 95 925 16379
LIB3061-017-Q1-K1-D7 LIB3061 g2198850 BLASTN 1183 1e-89 99 926
16379 700206765H1 SATMON003 g2198850 BLASTN 1095 1e-82 81 927 16379
700150281H1 SATMON007 g2198850 BLASTN 1024 1e-76 94 928 16379
700165862H1 SATMON013 g2198850 BLASTN 948 1e-75 94 929 2221
LIB3060-017-Q1-K1-B10 LIB3060 g2198850 BLASTN 1819 1e-161 99 930
2221 LIB84-006-Q1-E1-F3 LIB84 g2198852 BLASTN 1615 1e-153 97 931
2221 700575334H1 SATMON030 g2198850 BLASTN 1570 1e-124 99 932 2221
700206250H1 SATMON003 g2198850 BLASTN 1515 1e-117 100 933 2221
700095073H1 SATMON008 g2198852 BLASTN 1490 1e-115 100 934 2221
700571230H1 SATMON030 g2198850 BLASTN 1355 1e-114 93 935 2221
700157358H1 SATMON012 g2198850 BLASTN 1370 1e-105 100 936 2221
700379811H1 SATMON021 g2198850 BLASTN 1345 1e-103 93 937 2221
700041570H1 SATMON004 g2198850 BLASTN 1325 1e-101 100 938 2221
700104063H1 SATMON010 g2198850 BLASTN 1157 1e-95 90 939 2221
700378265H1 SATMON019 g2198850 BLASTN 771 1e-94 99 940 2221
700235329H1 SATMON010 g2198850 BLASTN 1234 1e-94 93 941 2221
LIB3068-057-Q1-K1-D1 LIB3068 g2198852 BLASTN 1209 1e-91 94 942 2221
700159158H1 SATMON012 g2198850 BLASTN 1174 1e-89 94 943 2221
700580854H1 SATMON031 g2198850 BLASTN 754 1e-86 92 944 2221
700623409H1 SATMON034 g2198850 BLASTN 910 1e-84 96 945 2221
701164706H1 SATMONN04 g2198852 BLASTN 577 1e-83 94 946 2221
700164719H1 SATMON013 g2198852 BLASTN 831 1e-83 99 947 2221
700158146H1 SATMON012 g2198850 BLASTN 1100 1e-82 93 948 2221
700203970H1 SATMON003 g2198852 BLASTN 996 1e-80 99 949 2221
700158313H1 SATMON012 g2198850 BLASTN 1036 1e-77 93 950 2221
700425211H1 SATMONN01 g2198852 BLASTN 485 1e-61 96 951 2221
700167764H1 SATMON013 g2198850 BLASTN 793 1e-57 93 952 23788
700102780H1 SATMON010 g2198852 BLASTN 1131 1e-104 99 953 23788
701167458H1 SATMONN05 g2198852 BLASTN 1069 1e-90 93 2655 -700900206
700900206H1 SOYMON027 g1742961 BLASTX 215 1e-24 78 2656 -GM40351
LIB3051-114-Q1-K1-H12 LIB3051 g2198851 BLASTX 122 1e-25 96 2657
12502 701101592H1 SOYMON028 g146846 BLASTX 103 1e-18 44 2658 12502
701106834H1 SOYMON036 g146846 BLASTX 103 1e-18 44 2659 13820
LIB3055-003-Q1-N1-F12 LIB3055 g3202028 BLASTX 193 1e-35 94 2660
8119 700989656H1 SOYMON011 g1742960 BLASTN 815 1e-59 79
CYSTATHIONINE .beta.-LYASE (EC 4.4.1.8) 954 -700155172 700155172H1
SATMON007 g704397 BLASTX 361 1e-43 78 955 -L362943
LIB36-013-Q1-E1-G10 LIB36 g704396 BLASTN 818 1e-59 75 956 19856
700240664H1 SATMON010 g704396 BLASTN 496 1e-31 69 957 19856
700572496H1 SATMON030 g704396 BLASTN 446 1e-26 68 958 22960
701170964H1 SATMONN05 g704396 BLASTN 778 1e-56 78 959 22960
LIB3061-002-Q1-K2-F9 LIB3061 g704396 BLASTN 765 1e-53 77 960 22960
701172780H2 SATMONN05 g704396 BLASTN 686 1e-48 73 961 22960
700578571H1 SATMON031 g704397 BLASTX 222 1e-23 89 962 30752
LIB3078-055-Q1-K1-C8 LIB3078 g704396 BLASTN 747 1e-51 71 963 30752
700086603H1 SATMON011 g704397 BLASTX 192 1e-18 64 2661 -701001147
701001147H1 SOYMON018 g704396 BLASTN 847 1e-61 78 2662 18602
700566066H1 SOYMON002 g704396 BLASTN 751 1e-57 79 2663 18602
700890955H1 SOYMON024 g704396 BLASTN 698 1e-49 77 2664 18602
700896865H1 SOYMON027 g704396 BLASTN 682 1e-48 77 2665 5144
LIB3050-006-Q1-E1-A9 LIB3050 g1399263 BLASTX 96 1e-31 41
5-METHYLTETRAHYDROPTEROYLTRIGLUTAMATE--HOMOCYSTEINE
S-METHYLTRANSFERASE
(EC 2.1.1.14) 964 -700212217 700212217H1 SATMON016 g886471 BLASTX
151 1e-16 93 965 -700333966 700333966H1 SATMON019 g886470 BLASTN
370 1e-23 66 966 -700377403 700377403H1 SATMON019 g2738247 BLASTN
305 1e-26 80 967 -700571893 700571893H1 SATMON030 g974781 BLASTN
262 1e-20 80 968 -701165656 701165656H1 SATMONN04 g2738247 BLASTN
450 1e-50 73 969 -L30622954 LIB3062-030-Q1-K1-C9 LIB3062 g886470
BLASTN 366 1e-26 81 970 -L30662004 LIB3066-026-Q1-K1-F1 LIB3066
g2738248 BLASTX 140 1e-28 77 971 -L30671450 LIB3067-001-Q1-K1-C6
LIB3067 g2738247 BLASTN 553 1e-46 69 972 -L30694410
LIB3069-057-Q1-K1-B4 LIB3069 g886470 BLASTN 447 1e-26 67 973 1
700452404H1 SATMON028 g453939 BLASTX 59 1e-17 94 974 13513
700049012H1 SATMON003 g1814402 BLASTN 1006 1e-74 81 975 13513
700086058H1 SATMON011 g1814402 BLASTN 982 1e-72 81 976 13513
700235814H1 SATMON010 g1814402 BLASTN 890 1e-65 79 977 13513
700170015H1 SATMON013 g1814402 BLASTN 578 1e-39 78 978 3835
700093161H1 SATMON008 g974781 BLASTN 858 1e-62 76 979 3835
700223321H1 SATMON011 g974781 BLASTN 836 1e-60 80 980 3835
700454003H1 SATMON029 g974781 BLASTN 823 1e-59 82 981 3835
700238925H1 SATMON010 g2738247 BLASTN 733 1e-55 74 982 3835
700075142H1 SATMON007 g2738247 BLASTN 569 1e-52 73 983 3835
700151969H1 SATMON007 g974781 BLASTN 660 1e-46 76 984 3835
700281434H2 SATMON019 g974781 BLASTN 560 1e-45 79 985 3835
700084914H1 SATMON011 g974781 BLASTN 440 1e-36 79 986 3835
700215135H1 SATMON016 g974781 BLASTN 543 1e-36 78 987 3835
700202218H1 SATMON003 g974781 BLASTN 389 1e-23 78 988 3835
700281467H2 SATMON019 g2738248 BLASTX 123 1e-12 71 989 456
LIB3059-019-Q1-K1-G6 LIB3059 g974781 BLASTN 1493 1e-115 83 990 456
LIB3068-012-Q1-K1-G7 LIB3068 g974781 BLASTN 1471 1e-113 81 991 456
LIB3061-050-Q1-K1-G10 LIB3061 g886470 BLASTN 1285 1e-98 82 992 456
LIB3067-043-Q1-K1-F9 LIB3067 g2738247 BLASTN 1287 1e-98 79 993 456
700087169H1 SATMON011 g1814402 BLASTN 1245 1e-94 86 994 456
LIB3069-030-Q1-K1-G9 LIB3069 g1814402 BLASTN 772 1e-93 80 995 456
LIB3069-019-Q1-K1-G11 LIB3069 g1814402 BLASTN 1202 1e-91 81 996 456
700202514H1 SATMON003 g1814402 BLASTN 1134 1e-89 84 997 456
LIB3062-011-Q1-K1-F2 LIB3062 g886470 BLASTN 664 1e-86 84 998 456
LIB143-015-Q1-E1-A1 LIB143 g974781 BLASTN 1143 1e-86 81 999 456
700570326H1 SATMON030 g974781 BLASTN 1089 1e-85 83 1000 456
700103727H1 SATMON010 g2738247 BLASTN 1132 1e-85 86 1001 456
700574823H1 SATMON030 g1814402 BLASTN 1135 1e-85 80 1002 456
700091666H1 SATMON011 g1814402 BLASTN 1136 1e-85 84 1003 456
LIB3069-033-Q1-K1-E9 LIB3069 g2738247 BLASTN 1115 1e-84 79 1004 456
700572634H1 SATMON030 g1814402 BLASTN 1123 1e-84 85 1005 456
700211829H1 SATMON016 g2738247 BLASTN 1096 1e-82 84 1006 456
700087109H1 SATMON011 g1814402 BLASTN 1102 1e-82 85 1007 456
LIB3059-003-Q1-K1-B1 LIB3059 g1814402 BLASTN 961 1e-81 82 1008 456
700047556H1 SATMON003 g2738247 BLASTN 976 1e-81 82 1009 456
700201925H1 SATMON003 g2738247 BLASTN 1001 1e-81 84 1010 456
700209343H1 SATMON016 g1814402 BLASTN 1081 1e-81 82 1011 456
700348416H1 SATMON023 g1814402 BLASTN 1081 1e-81 83 1012 456
700095559H1 SATMON008 g1814402 BLASTN 1085 1e-81 82 1013 456
700073472H1 SATMON007 g1814402 BLASTN 1089 1e-81 82 1014 456
LIB3062-057-Q1-K1-C2 LIB3062 g886470 BLASTN 1073 1e-80 78 1015 456
700093445H1 SATMON008 g1814402 BLASTN 1076 1e-80 84 1016 456
LIB143-026-Q1-E1-H3 LIB143 g2738247 BLASTN 648 1e-79 79 1017 456
700206240H1 SATMON003 g1814402 BLASTN 1057 1e-79 82 1018 456
700091907H1 SATMON011 g886470 BLASTN 1062 1e-79 80 1019 456
700258178H1 SATMON017 g1814402 BLASTN 1064 1e-79 84 1020 456
700086565H1 SATMON011 g1814402 BLASTN 1065 1e-79 82 1021 456
700471452H1 SATMON025 g2738247 BLASTN 1045 1e-78 85 1022 456
700243858H1 SATMON010 g1814402 BLASTN 1048 1e-78 85 1023 456
700352224H1 SATMON023 g2738247 BLASTN 1051 1e-78 84 1024 456
700084769H1 SATMON011 g1814402 BLASTN 1054 1e-78 83 1025 456
700082818H1 SATMON011 g886470 BLASTN 984 1e-77 84 1026 456
700331974H1 SATMON019 g886470 BLASTN 1031 1e-77 82 1027 456
700086203H1 SATMON011 g1814402 BLASTN 1032 1e-77 81 1028 456
700075826H1 SATMON007 g2738247 BLASTN 1038 1e-77 82 1029 456
700104555H1 SATMON010 g886470 BLASTN 1039 1e-77 79 1030 456
700074084H1 SATMON007 g2738247 BLASTN 1039 1e-77 87 1031 456
700076872H1 SATMON007 g1814402 BLASTN 700 1e-76 83 1032 456
700050704H1 SATMON003 g1814402 BLASTN 880 1e-76 82 1033 456
700030215H1 SATMON003 g974781 BLASTN 1019 1e-76 83 1034 456
700077416H1 SATMON007 g886470 BLASTN 1019 1e-76 79 1035 456
700093982H1 SATMON008 g1814402 BLASTN 1019 1e-76 82 1036 456
700048847H1 SATMON003 g2738247 BLASTN 1023 1e-76 85 1037 456
700090739H1 SATMON011 g886470 BLASTN 1026 1e-76 82 1038 456
700092904H1 SATMON008 g886470 BLASTN 1026 1e-76 82 1039 456
LIB3068-009-Q1-K1-E5 LIB3068 g1814402 BLASTN 1027 1e-76 73 1040 456
700336705H1 SATMON019 g1814402 BLASTN 747 1e-75 83 1041 456
700344872H1 SATMON021 g1814402 BLASTN 768 1e-75 85 1042 456
LIB3066-019-Q1-K1-E1 LIB3066 g886470 BLASTN 868 1e-75 81 1043 456
700086322H1 SATMON011 g2738247 BLASTN 1015 1e-75 81 1044 456
LIB3067-052-Q1-K1-B12 LIB3067 g974781 BLASTN 1033 1e-75 81 1045 456
700221395H1 SATMON011 g2738247 BLASTN 1000 1e-74 85 1046 456
700092483H1 SATMON008 g1814402 BLASTN 1001 1e-74 82 1047 456
700618708H1 SATMON034 g886470 BLASTN 1003 1e-74 82 1048 456
700074191H1 SATMON007 g2738247 BLASTN 1005 1e-74 86 1049 456
LIB143-055-Q1-E1-F10 LIB143 g1814402 BLASTN 986 1e-73 83 1050 456
700025955H1 SATMON003 g1814402 BLASTN 993 1e-73 84 1051 456
700574824H1 SATMON030 g886470 BLASTN 564 1e-72 80 1052 456
700201527H1 SATMON003 g974781 BLASTN 812 1e-72 81 1053 456
700092202H1 SATMON008 g886470 BLASTN 946 1e-72 79 1054 456
700223674H1 SATMON011 g2738247 BLASTN 971 1e-72 83 1055 456
700092638H1 SATMON008 g1814402 BLASTN 972 1e-72 84 1056 456
700090025H1 SATMON011 g974781 BLASTN 973 1e-72 81 1057 456
700217069H1 SATMON016 g2738247 BLASTN 976 1e-72 83 1058 456
700349142H1 SATMON023 g1814402 BLASTN 980 1e-72 82 1059 456
700088387H1 SATMON011 g2738247 BLASTN 980 1e-72 85 1060 456
700215102H1 SATMON016 g886470 BLASTN 980 1e-72 79 1061 456
700456708H1 SATMON029 g2738247 BLASTN 982 1e-72 82 1062 456
700210025H1 SATMON016 g1814402 BLASTN 982 1e-72 81 1063 456
700335345H1 SATMON019 g974781 BLASTN 982 1e-72 84 1064 456
700085257H1 SATMON011 g1814402 BLASTN 554 1e-71 82 1065 456
700026781H1 SATMON003 g974781 BLASTN 961 1e-71 82 1066 456
700224265H1 SATMON011 g2738247 BLASTN 962 1e-71 83 1067 456
700083282H1 SATMON011 g1814402 BLASTN 962 1e-71 83 1068 456
700381324H1 SATMON023 g1814402 BLASTN 965 1e-71 82 1069 456
700212017H1 SATMON016 g1814402 BLASTN 640 1e-70 83 1070 456
700088053H1 SATMON011 g886470 BLASTN 853 1e-70 84 1071 456
LIB3062-032-Q1-K1-C2 LIB3062 g1814402 BLASTN 883 1e-70 80 1072 456
700083030H1 SATMON011 g1814402 BLASTN 948 1e-70 81 1073 456
700613980H1 SATMON033 g1814402 BLASTN 950 1e-70 83 1074 456
700347019H1 SATMON021 g974781 BLASTN 952 1e-70 81 1075 456
700335621H1 SATMON019 g1814402 BLASTN 952 1e-70 80 1076 456
700094101H1 SATMON008 g1814402 BLASTN 953 1e-70 83 1077 456
700073128H1 SATMON007 g2738247 BLASTN 954 1e-70 78 1078 456
700076245H1 SATMON007 g1814402 BLASTN 955 1e-70 83 1079 456
700071903H1 SATMON007 g886470 BLASTN 956 1e-70 81 1080 456
700090035H1 SATMON011 g886470 BLASTN 935 1e-69 80 1081 456
700405323H1 SATMON029 g1814402 BLASTN 937 1e-69 83 1082 456
700050121H1 SATMON003 g886470 BLASTN 938 1e-69 83 1083 456
700224714H1 SATMON011 g1814402 BLASTN 939 1e-69 82 1084 456
700281535H2 SATMON019 g1814402 BLASTN 940 1e-69 81 1085 456
700222093H1 SATMON011 g2738247 BLASTN 941 1e-69 81 1086 456
700094659H1 SATMON008 g1814402 BLASTN 942 1e-69 84 1087 456
700237187H1 SATMON010 g1814402 BLASTN 942 1e-69 84 1088 456
700023210H1 SATMON003 g1814402 BLASTN 943 1e-69 83 1089 456
700072813H1 SATMON007 g1814402 BLASTN 944 1e-69 82 1090 456
700075720H1 SATMON007 g1814402 BLASTN 946 1e-69 83 1091 456
700072107H1 SATMON007 g1814402 BLASTN 946 1e-69 80 1092 456
700082225H1 SATMON011 g1814402 BLASTN 691 1e-68 82 1093 456
700085411H1 SATMON011 g1814402 BLASTN 723 1e-68 83 1094 456
700454357H1 SATMON029 g2738247 BLASTN 926 1e-68 82 1095 456
700620976H1 SATMON034 g1814402 BLASTN 927 1e-68 81 1096 456
700241483H1 SATMON010 g1814402 BLASTN 931 1e-68 86 1097 456
700355459H1 SATMON024 g974781 BLASTN 931 1e-68 82 1098 456
700095875H1 SATMON008 g1814402 BLASTN 931 1e-68 87 1099 456
700076702H1 SATMON007 g1814402 BLASTN 934 1e-68 84 1100 456
700212538H1 SATMON016 g1814402 BLASTN 689 1e-67 85 1101 456
700086479H1 SATMON011 g1814402 BLASTN 715 1e-67 82 1102 456
700453684H1 SATMON028 g1814402 BLASTN 747 1e-67 83 1103 456
700204454H1 SATMON003 g1814402 BLASTN 797 1e-67 82 1104 456
700612577H1 SATMON033 g974781 BLASTN 911 1e-67 81 1105 456
700474018H1 SATMON025 g886470 BLASTN 917 1e-67 81 1106 456
700213602H1 SATMON016 g886470 BLASTN 917 1e-67 76 1107 456
700076911H1 SATMON007 g974781 BLASTN 917 1e-67 81 1108 456
700223313H1 SATMON011 g886470 BLASTN 919 1e-67 84 1109 456
700220692H1 SATMON011 g974781 BLASTN 920 1e-67 81 1110 456
700258038H1 SATMON017 g886470 BLASTN 920 1e-67 83 1111 456
700095862H1 SATMON008 g1814402 BLASTN 922 1e-67 86 1112 456
700213396H1 SATMON016 g886470 BLASTN 922 1e-67 83 1113 456
700354259H1 SATMON024 g974781 BLASTN 638 1e-66 81 1114 456
700471505H1 SATMON025 g886470 BLASTN 681 1e-66 83 1115 456
700202439H1 SATMON003 g1814402 BLASTN 706 1e-66 82 1116 456
700223609H1 SATMON011 g974781 BLASTN 770 1e-66 83 1117 456
700405260H1 SATMON028 g1814402 BLASTN 782 1e-66 86 1118 456
700085214H1 SATMON011 g886470 BLASTN 814 1e-66 84 1119 456
700211194H1 SATMON016 g1814402 BLASTN 902 1e-66 84 1120 456
700106641H1 SATMON010 g1814402 BLASTN 903 1e-66 82 1121 456
700405170H1 SATMON028 g974781 BLASTN 904 1e-66 79 1122 456
700090015H1 SATMON011 g886470 BLASTN 905 1e-66 80 1123 456
700623154H1 SATMON034 g886470 BLASTN 906 1e-66 81 1124 456
700458895H1 SATMON029 g886470 BLASTN 906 1e-66 83 1125 456
700094073H1 SATMON008 g886470 BLASTN 907 1e-66 78 1126 456
700158508H1 SATMON012 g2738247 BLASTN 909 1e-66 82 1127 456
700027366H1 SATMON003 g1814402 BLASTN 910 1e-66 81 1128 456
700073636H1 SATMON007 g1814402 BLASTN 615 1e-65 87 1129 456
700220207H1 SATMON011 g886470 BLASTN 785 1e-65 84 1130 456
LIB143-007-Q1-E1-A6 LIB143 g886470 BLASTN 807 1e-65 80 1131 456
700575288H1 SATMON030 g1814402 BLASTN 887 1e-65 83 1132 456
700219870H1 SATMON011 g974781 BLASTN 887 1e-65 82 1133 456
701184573H1 SATMONN06 g974781 BLASTN 888 1e-65 81 1134 456
700076946H1 SATMON007 g1814402 BLASTN 891 1e-65 86 1135 456
700214808H1 SATMON016 g2738247 BLASTN 893 1e-65 81 1136 456
700096274H1 SATMON008 g1814402 BLASTN 894 1e-65 80 1137 456
700160445H1 SATMON012 g1814402 BLASTN 896 1e-65 85 1138 456
700071727H1 SATMON007 g974781 BLASTN 898 1e-65 80 1139 456
700571751H1 SATMON030 g1814402 BLASTN 898 1e-65 78 1140 456
700215107H1 SATMON016 g2738247 BLASTN 507 1e-64 77 1141 456
LIB3067-057-Q1-K1-C6 LIB3067 g2738247 BLASTN 555 1e-64 76 1142 456
700614743H1 SATMON033 g886470 BLASTN 748 1e-64 81 1143 456
700444971H1 SATMON027 g974781 BLASTN 803 1e-64 83 1144 456
700224573H1 SATMON011 g1814402 BLASTN 805 1e-64 86 1145 456
700019276H1 SATMON001 g1814402 BLASTN 875 1e-64 83 1146 456
700210440H1 SATMON016 g2738247 BLASTN 881 1e-64 80 1147 456
700093168H1 SATMON008 g1814402 BLASTN 885 1e-64 86 1148 456
700224478H1 SATMON011 g886470 BLASTN 886 1e-64 81 1149 456
700261958H1 SATMON017 g1814402 BLASTN 781 1e-63 79 1150 456
700074221H1 SATMON007 g1814402 BLASTN 825 1e-63 81 1151 456
700089966H1 SATMON011 g1814402 BLASTN 863 1e-63 84 1152 456
700210422H1 SATMON016 g1814402 BLASTN 863 1e-63 81 1153 456
700160272H1 SATMON012 g1814402 BLASTN 863 1e-63 85 1154 456
700444282H1 SATMON027 g2738247 BLASTN 549 1e-62 84 1155 456
700206489H1 SATMON003 g2738247 BLASTN 597 1e-62 80 1156 456
700446452H1 SATMON027 g974781 BLASTN 711 1e-62 80 1157 456
700265473H1 SATMON017 g1814402 BLASTN 715 1e-62 79 1158 456
700473708H1 SATMON025 g1814402 BLASTN 762 1e-62 82 1159 456
LIB3069-025-Q1-K1-A7 LIB3069 g886470 BLASTN 852 1e-62 80 1160 456
700166631H1 SATMON013 g2738247 BLASTN 858 1e-62 85 1161 456
700582413H1 SATMON031 g1814402 BLASTN 861 1e-62 78 1162 456
700071904H1 SATMON007 g2738247 BLASTN 544 1e-61 77 1163 456
700622655H1 SATMON034 g1814402 BLASTN 592 1e-61 82 1164 456
700082541H1 SATMON011 g1814402 BLASTN 793 1e-61 82 1165 456
700335378H1 SATMON019 g2738247 BLASTN 841 1e-61 76 1166 456
700203585H1 SATMON003 g2738247 BLASTN 846 1e-61 86 1167 456
700053043H1 SATMON007 g2738247 BLASTN 847 1e-61 81 1168 456
700163654H1 SATMON013 g1814402 BLASTN 474 1e-60 84 1169 456
700352269H1 SATMON023 g1814402 BLASTN 479 1e-60 84 1170 456
700106187H1 SATMON010 g1814402 BLASTN 631 1e-60 81 1171 456
LIB143-007-Q1-E1-A7 LIB143 g886470 BLASTN 759 1e-60 76 1172 456
700458694H1 SATMON029 g1814402 BLASTN 827 1e-60 86 1173 456
700213509H1 SATMON016 g886470 BLASTN 834 1e-60 81 1174 456
700150191H1 SATMON007 g2738247 BLASTN 834 1e-60 82 1175 456
700157594H1 SATMON012 g1814402 BLASTN 835 1e-60 80 1176 456
700094046H1 SATMON008 g974781 BLASTN 836 1e-60 79 1177 456
700220254H1 SATMON011 g2738247 BLASTN 687 1e-59 77 1178 456
700095485H1 SATMON008 g886470 BLASTN 819 1e-59 77 1179 456
LIB143-006-Q1-E1-B3 LIB143 g1814402 BLASTN 830 1e-59 65 1180 456
700071763H1 SATMON007 g2738247 BLASTN 565 1e-58 75 1181 456
700082259H1 SATMON011 g1814402 BLASTN 707 1e-58 84 1182 456
700020717H1 SATMON001 g886470 BLASTN 805 1e-58 81 1183 456
700041892H1 SATMON004 g1814402 BLASTN 805 1e-58 86 1184 456
700356573H1 SATMON024 g1814402 BLASTN 811 1e-58 81 1185 456
700171632H1 SATMON013 g2738247 BLASTN 811 1e-58 85 1186 456
700222516H1 SATMON011 g1814402 BLASTN 812 1e-58 87 1187 456
700466679H1 SATMON025 g1814402 BLASTN 812 1e-58 87 1188 456
700208929H1 SATMON016 g1814402 BLASTN 407 1e-57 78 1189 456
700355014H1 SATMON024 g1814402 BLASTN 592 1e-57 85 1190 456
701159834H1 SATMONN04 g1814402 BLASTN 683 1e-57 77 1191 456
700351137H1 SATMON023 g974781 BLASTN 725 1e-57 79 1192 456
700027308H1 SATMON003 g2738247 BLASTN 791 1e-57 80 1193 456
700142587H1 SATMON012 g974781 BLASTN 791 1e-57 83 1194 456
700170157H1 SATMON013 g1814402 BLASTN 791 1e-57 81 1195 456
700019438H1 SATMON001 g974781 BLASTN 798 1e-57 82 1196 456
700166430H1 SATMON013 g886470 BLASTN 799 1e-57 84 1197 456
700047369H1 SATMON003 g886470 BLASTN 799 1e-57 80 1198 456
700152373H1 SATMON007 g2738247 BLASTN 799 1e-57 84 1199 456
700050824H1 SATMON003 g2738247 BLASTN 605 1e-56 81 1200 456
700204588H1 SATMON003 g886470 BLASTN 720 1e-56 82 1201 456
700071667H1 SATMON007 g1814402 BLASTN 787 1e-56 85 1202 456
701185269H1 SATMONN06 g886470 BLASTN 598 1e-55 76 1203 456
700243844H1 SATMON010 g2738247 BLASTN 769 1e-55 79 1204 456
700221969H1 SATMON011 g974781 BLASTN 771 1e-55 79 1205 456
700335706H1 SATMON019 g1814402 BLASTN 773 1e-55 80 1206 456
700622285H1 SATMON034 g1814402 BLASTN 774 1e-55 82 1207 456
700089665H1 SATMON011 g2738247 BLASTN 774 1e-55 78 1208 456
700464833H1 SATMON025 g2738247 BLASTN 455 1e-54 72 1209 456
LIB3069-057-Q1-K1-A6 LIB3069 g974781 BLASTN 564 1e-54 76 1210 456
700156818H1 SATMON012 g974781 BLASTN 757 1e-54 81 1211 456
700219420H1 SATMON011 g974781 BLASTN 758 1e-54 79 1212 456
700152570H1 SATMON007 g1814402 BLASTN 761 1e-54 86 1213 456
700084484H1 SATMON011 g1814402 BLASTN 764 1e-54 84
1214 456 700152208H1 SATMON007 g1814402 BLASTN 765 1e-54 82 1215
456 700550889H1 SATMON022 g2738247 BLASTN 546 1e-53 76 1216 456
700025869H1 SATMON003 g2738247 BLASTN 675 1e-53 79 1217 456
700383075H1 SATMON024 g974781 BLASTN 724 1e-53 81 1218 456
700575104H1 SATMON030 g1814402 BLASTN 745 1e-53 70 1219 456
LIB3069-022-Q1-K1-C1 LIB3069 g974781 BLASTN 768 1e-53 81 1220 456
700215272H1 SATMON016 g1814402 BLASTN 674 1e-52 83 1221 456
700167881H1 SATMON013 g974781 BLASTN 737 1e-52 79 1222 456
700019727H1 SATMON001 g2738247 BLASTN 737 1e-52 81 1223 456
700351776H1 SATMON023 g886470 BLASTN 741 1e-52 76 1224 456
700210543H1 SATMON016 g974781 BLASTN 742 1e-52 85 1225 456
700571604H1 SATMON030 g1814402 BLASTN 360 1e-51 76 1226 456
700169777H1 SATMON013 g1814402 BLASTN 426 1e-51 83 1227 456
700096744H1 SATMON008 g2738247 BLASTN 539 1e-51 79 1228 456
700383157H1 SATMON024 g974781 BLASTN 667 1e-51 79 1229 456
700155764H1 SATMON007 g886470 BLASTN 721 1e-51 75 1230 456
700150817H1 SATMON007 g1814402 BLASTN 727 1e-51 75 1231 456
700619660H1 SATMON034 g886470 BLASTN 727 1e-51 73 1232 456
700471085H1 SATMON025 g1814402 BLASTN 406 1e-50 79 1233 456
700163226H1 SATMON013 g2738247 BLASTN 475 1e-50 78 1234 456
700444567H1 SATMON027 g1814402 BLASTN 498 1e-50 81 1235 456
700457303H1 SATMON029 g2738247 BLASTN 662 1e-50 80 1236 456
700088065H1 SATMON011 g886470 BLASTN 707 1e-50 79 1237 456
700152592H1 SATMON007 g886470 BLASTN 716 1e-50 80 1238 456
LIB3068-061-Q1-K1-B2 LIB3068 g886470 BLASTN 730 1e-50 78 1239 456
700331880H1 SATMON019 g1814402 BLASTN 697 1e-49 85 1240 456
700218025H1 SATMON016 g886470 BLASTN 700 1e-49 79 1241 456
700348643H1 SATMON023 g1814402 BLASTN 701 1e-49 80 1242 456
700236975H1 SATMON010 g974781 BLASTN 704 1e-49 81 1243 456
700442968H1 SATMON026 g886470 BLASTN 418 1e-48 79 1244 456
700479529H1 SATMON034 g2738247 BLASTN 508 1e-48 75 1245 456
700160983H1 SATMON012 g1814402 BLASTN 690 1e-48 83 1246 456
700050185H1 SATMON003 g974781 BLASTN 421 1e-47 75 1247 456
700611764H1 SATMON022 g1814402 BLASTN 503 1e-47 79 1248 456
700622669H1 SATMON034 g1814402 BLASTN 549 1e-47 79 1249 456
700153237H1 SATMON007 g1814402 BLASTN 672 1e-47 80 1250 456
700242703H1 SATMON010 g886470 BLASTN 682 1e-47 81 1251 456
700165177H1 SATMON013 g2738247 BLASTN 682 1e-47 78 1252 456
700379889H1 SATMON021 g886470 BLASTN 682 1e-47 85 1253 456
700449243H1 SATMON028 g1814402 BLASTN 357 1e-46 83 1254 456
700453282H1 SATMON028 g886470 BLASTN 608 1e-46 81 1255 456
700224308H1 SATMON011 g1814402 BLASTN 661 1e-46 83 1256 456
700150484H1 SATMON007 g886470 BLASTN 663 1e-46 78 1257 456
700240609H1 SATMON010 g2738247 BLASTN 668 1e-46 79 1258 456
700171610H1 SATMON013 g974781 BLASTN 669 1e-46 79 1259 456
LIB143-027-Q1-E1-H3 LIB143 g1814402 BLASTN 670 1e-46 86 1260 456
700334084H1 SATMON019 g1814402 BLASTN 467 1e-45 81 1261 456
LIB3069-043-Q1-K1-H6 LIB3069 g886470 BLASTN 546 1e-45 83 1262 456
700222515H1 SATMON011 g1814402 BLASTN 648 1e-45 75 1263 456
700223749H1 SATMON011 g1814402 BLASTN 648 1e-45 75 1264 456
701180187H1 SATMONN05 g974781 BLASTN 648 1e-45 78 1265 456
700050111H1 SATMON003 g2738247 BLASTN 653 1e-45 82 1266 456
700051275H1 SATMON003 g1814402 BLASTN 379 1e-44 75 1267 456
700235202H1 SATMON010 g2738247 BLASTN 637 1e-44 80 1268 456
700257385H1 SATMON017 g1814402 BLASTN 638 1e-44 80 1269 456
LIB143-028-Q1-E1-H7 LIB143 g1814402 BLASTN 638 1e-44 80 1270 456
700576066H1 SATMON030 g886470 BLASTN 516 1e-43 76 1271 456
700455065H1 SATMON029 g886470 BLASTN 624 1e-43 79 1272 456
700074158H1 SATMON007 g1814402 BLASTN 630 1e-43 83 1273 456
700171322H1 SATMON013 g1814402 BLASTN 634 1e-43 77 1274 456
700457369H1 SATMON029 g974781 BLASTN 392 1e-42 83 1275 456
700378252H1 SATMON019 g2738247 BLASTN 462 1e-42 77 1276 456
700454626H1 SATMON029 g1814402 BLASTN 495 1e-42 85 1277 456
700208182H1 SATMON016 g2738247 BLASTN 564 1e-42 73 1278 456
700618848H1 SATMON034 g886470 BLASTN 615 1e-42 80 1279 456
700222969H1 SATMON011 g1814402 BLASTN 622 1e-42 74 1280 456
700444782H1 SATMON027 g1814402 BLASTN 337 1e-41 81 1281 456
700204404H1 SATMON003 g2738247 BLASTN 607 1e-41 80 1282 456
700347028H1 SATMON021 g1814402 BLASTN 598 1e-40 81 1283 456
700172447H1 SATMON013 g1814402 BLASTN 328 1e-39 85 1284 456
700549696H1 SATMON022 g886470 BLASTN 360 1e-39 81 1285 456
700257287H1 SATMON017 g1814402 BLASTN 365 1e-38 84 1286 456
700569630H1 SATMON030 g1814402 BLASTN 566 1e-38 78 1287 456
700207258H1 SATMON017 g1814402 BLASTN 568 1e-38 85 1288 456
700052467H1 SATMON003 g1814402 BLASTN 515 1e-37 82 1289 456
700083656H1 SATMON011 g2738247 BLASTN 552 1e-37 83 1290 456
700429279H1 SATMONN01 g2738247 BLASTN 556 1e-37 79 1291 456
700349921H1 SATMON023 g2738247 BLASTN 471 1e-36 78 1292 456
700449609H1 SATMON028 g886470 BLASTN 540 1e-36 81 1293 456
700150152H1 SATMON007 g1814402 BLASTN 543 1e-36 74 1294 456
700075675H1 SATMON007 g1814402 BLASTN 546 1e-36 81 1295 456
700267015H1 SATMON017 g886470 BLASTN 550 1e-36 75 1296 456
700465118H1 SATMON025 g1814402 BLASTN 555 1e-36 78 1297 456
700456662H1 SATMON029 g2738247 BLASTN 314 1e-34 76 1298 456
700405315H1 SATMON029 g886470 BLASTN 519 1e-34 83 1299 456
700218639H1 SATMON011 g974781 BLASTN 524 1e-34 81 1300 456
700216606H1 SATMON016 g1814402 BLASTN 524 1e-34 83 1301 456
700456965H1 SATMON029 g2738247 BLASTN 525 1e-34 78 1302 456
700102147H1 SATMON010 g974781 BLASTN 526 1e-34 82 1303 456
700235734H1 SATMON010 g2738247 BLASTN 526 1e-34 83 1304 456
LIB3068-009-Q1-K1-E10 LIB3068 g886470 BLASTN 548 1e-34 70 1305 456
700029363H1 SATMON003 g974781 BLASTN 377 1e-33 80 1306 456
LIB3069-030-Q1-K1-D10 LIB3069 g886470 BLASTN 297 1e-30 78 1307 456
700334895H1 SATMON019 g974781 BLASTN 468 1e-30 82 1308 456
700150963H1 SATMON007 g974781 BLASTN 459 1e-29 88 1309 456
700152817H1 SATMON007 g974781 BLASTN 459 1e-29 88 1310 456
700208732H1 SATMON016 g974781 BLASTN 485 1e-29 76 1311 456
700051461H1 SATMON003 g886470 BLASTN 467 1e-28 73 1312 456
700616589H1 SATMON033 g2738247 BLASTN 273 1e-27 78 1313 456
700236150H1 SATMON010 g974781 BLASTN 434 1e-27 84 1314 456
700453369H1 SATMON028 g1814402 BLASTN 436 1e-27 84 1315 456
700621739H1 SATMON034 g1814402 BLASTN 436 1e-27 69 1316 456
700075162H1 SATMON007 g1814402 BLASTN 438 1e-27 85 1317 456
700405040H1 SATMON027 g974781 BLASTN 438 1e-27 72 1318 456
700356893H1 SATMON024 g974781 BLASTN 458 1e-27 82 1319 456
701185493H1 SATMONN06 g886471 BLASTX 72 1e-26 97 1320 456
700206095H1 SATMON003 g974781 BLASTN 421 1e-26 83 1321 456
700083901H1 SATMON011 g1814402 BLASTN 421 1e-26 80 1322 456
LIB3062-026-Q1-K1-D7 LIB3062 g1814402 BLASTN 423 1e-24 81 1323 456
700377277H1 SATMON019 g1814402 BLASTN 357 1e-20 75 1324 456
700201214H1 SATMON003 g2738248 BLASTX 152 1e-18 77 1325 456
700025959H1 SATMON003 g886471 BLASTX 188 1e-18 94 1326 456
700259484H1 SATMON017 g1814402 BLASTN 189 1e-17 82 1327 456
700613969H1 SATMON033 g886470 BLASTN 315 1e-17 90 1328 456
700215602H1 SATMON016 g2738248 BLASTX 158 1e-16 74 1329 456
700438152H1 SATMON026 g2738247 BLASTN 258 1e-16 75 1330 456
700458654H1 SATMON029 g886470 BLASTN 330 1e-16 70 1331 456
700449563H1 SATMON028 g886471 BLASTX 163 1e-15 89 1332 456
700236490H1 SATMON010 g1814402 BLASTN 276 1e-14 89 1333 456
700456927H1 SATMON029 g1814403 BLASTX 119 1e-13 86 1334 456
701180088H1 SATMONN05 g886471 BLASTX 152 1e-13 60 1335 456
700455716H1 SATMON029 g2738248 BLASTX 111 1e-12 67 1336 456
700573520H1 SATMON030 g886471 BLASTX 82 1e-11 84 1337 456
700569938H1 SATMON030 g2738248 BLASTX 132 1e-11 91 1338 456
700551958H1 SATMON022 g2738247 BLASTN 264 1e-11 82 1339 456
700337475H1 SATMON020 g2738247 BLASTN 268 1e-11 83 1340 456
700170827H1 SATMON013 g886471 BLASTX 124 1e-10 77 1341 456
700453382H1 SATMON028 g974781 BLASTN 152 1e-10 85 1342 456
700089211H1 SATMON011 g2738247 BLASTN 245 1e-9 79 1343 456
700103155H1 SATMON010 g2738248 BLASTX 82 1e-8 85 1344 5523
LIB3062-017-Q1-K1-F4 LIB3062 g2738247 BLASTN 932 1e-84 76 1345 5523
700210708H1 SATMON016 g2738247 BLASTN 952 1e-70 79 1346 5523
700219307H1 SATMON011 g1814402 BLASTN 892 1e-65 78 1347 5523
700221188H1 SATMON011 g2738247 BLASTN 867 1e-63 81 1348 5523
700203884H1 SATMON003 g2738247 BLASTN 874 1e-63 77 1349 5523
700218549H1 SATMON011 g2738247 BLASTN 811 1e-58 77 1350 5523
700572845H2 SATMON030 g2738247 BLASTN 785 1e-56 77 1351 5523
700152362H1 SATMON007 g886470 BLASTN 742 1e-52 79 1352 5523
700152138H1 SATMON007 g2738247 BLASTN 714 1e-50 80 1353 5523
700218842H1 SATMON011 g2738247 BLASTN 700 1e-49 75 2666 -700697958
700697958H1 SOYMON015 g2738247 BLASTN 258 1e-10 83 2667 -700731277
700731277H1 SOYMON009 g886470 BLASTN 930 1e-68 83 2668 -700749261
700749261H1 SOYMON013 g1814402 BLASTN 508 1e-38 78 2669 -700787742
700787742H2 SOYMON011 g1814402 BLASTN 648 1e-50 81 2670 -700831146
700831146H1 SOYMON019 g1814402 BLASTN 476 1e-34 80 2671 -700832029
700832029H1 SOYMON019 g1814402 BLASTN 328 1e-27 78 2672 -700854481
700854481H1 SOYMON023 g1814402 BLASTN 193 1e-11 85 2673 -700873716
700873716H1 SOYMON018 g1749542 BLASTX 99 1e-15 53 2674 -700893904
700893904H1 SOYMON024 g886470 BLASTN 413 1e-25 82 2675 -700909658
700909658H1 SOYMON022 g1814402 BLASTN 251 1e-10 91 2676 -700943841
700943841H1 SOYMON024 g886470 BLASTN 549 1e-49 82 2677 -700963223
700963223H1 SOYMON022 g886470 BLASTN 649 1e-45 72 2678 -700974103
700974103H1 SOYMON005 g886470 BLASTN 763 1e-54 80 2679 -700994293
700994293H1 SOYMON011 g974782 BLASTX 97 1e-12 72 2680 -701004816
701004816H1 SOYMON019 g1814402 BLASTN 582 1e-39 84 2681 -701007125
701007125H1 SOYMON019 g1814403 BLASTX 152 1e-13 87 2682 -701008540
701008540H1 SOYMON019 g886470 BLASTN 770 1e-55 74 2683 -701037766
701037766H1 SOYMON029 g1814403 BLASTX 72 1e-10 69 2684 -701062191
701062191H1 SOYMON033 g974781 BLASTN 225 1e-9 79 2685 -701105474
701105474H1 SOYMON036 g974782 BLASTX 164 1e-15 91 2686 -GM14442
LIB3049-056-Q1-E1-B1 LIB3049 g974781 BLASTN 231 1e-10 79 2687
-GM19631 LIB3056-007-Q1-N1-G9 LIB3056 g974781 BLASTN 388 1e-37 78
2688 -GM37189 LIB3051-072-Q1-K1-B5 LIB3051 g1814402 BLASTN 316
1e-15 78 2689 -GM44802 LIB3053-004-Q1-N1-B2 LIB3053 g974782 BLASTX
85 1e-26 90 2690 1382 700683236H1 SOYMON008 g886470 BLASTN 805
1e-58 78 2691 1382 700566434H1 SOYMON002 g886471 BLASTX 162 1e-23
76 2692 15690 701064361H1 SOYMON034 g974781 BLASTN 951 1e-70 84
2693 15690 700847301H1 SOYMON021 g886470 BLASTN 707 1e-66 85 2694
17335 LIB3051-088-Q1-K1-D9 LIB3051 g886470 BLASTN 1285 1e-101 79
2695 17335 701003832H1 SOYMON019 g974781 BLASTN 811 1e-58 77 2696
17335 700864888H1 SOYMON016 g1814402 BLASTN 767 1e-55 79 2697 17335
700672491H1 SOYMON006 g974781 BLASTN 428 1e-46 75 2698 17335
701003457H1 SOYMON019 g1814402 BLASTN 411 1e-29 84 2699 17335
700833672H1 SOYMON019 g974781 BLASTN 445 1e-28 79 2700 17900
700850578H1 SOYMON023 g2738247 BLASTN 905 1e-66 82 2701 17900
701053154H1 SOYMON032 g1814402 BLASTN 725 1e-65 83 2702 17900
700842351H1 SOYMON020 g1814402 BLASTN 878 1e-64 83 2703 17900
700837656H1 SOYMON020 g1814402 BLASTN 841 1e-61 83 2704 17900
700890851H1 SOYMON024 g974781 BLASTN 747 1e-53 80 2705 20688
700908840H1 SOYMON022 g2738247 BLASTN 889 1e-65 82 2706 20688
700908848H1 SOYMON022 g2738247 BLASTN 877 1e-64 81 2707 33542
LIB3051-009-Q1-E1-E5 LIB3051 g974781 BLASTN 1218 1e-92 79 2708
33542 700748773H1 SOYMON013 g974781 BLASTN 669 1e-46 78 2709 33542
700836363H1 SOYMON020 g2738247 BLASTN 596 1e-40 80 2710 4243
701123616H1 SOYMON037 g1814402 BLASTN 995 1e-74 87 2711 4243
700555001H1 SOYMON001 g1814402 BLASTN 975 1e-72 90 2712 4243
701002967H1 SOYMON019 g1814402 BLASTN 950 1e-70 86 2713 4243
700653509H1 SOYMON003 g1814402 BLASTN 539 1e-69 85 2714 4243
701206028H1 SOYMON035 g1814402 BLASTN 938 1e-69 86 2715 4243
700962115H1 SOYMON022 g1814402 BLASTN 923 1e-68 86 2716 4243
700866243H1 SOYMON016 g1814402 BLASTN 909 1e-66 82 2717 4243
700752507H1 SOYMON014 g1814402 BLASTN 887 1e-65 85 2718 4243
701003887H1 SOYMON019 g1814402 BLASTN 863 1e-63 86 2719 4243
700556913H1 SOYMON001 g1814402 BLASTN 865 1e-63 86 2720 4243
701013549H1 SOYMON019 g1814402 BLASTN 867 1e-63 90 2721 4243
701209706H1 SOYMON035 g1814402 BLASTN 871 1e-63 90 2722 4243
701010487H1 SOYMON019 g1814402 BLASTN 529 1e-62 79 2723 4243
700548246H1 SOYMON002 g1814402 BLASTN 553 1e-62 82 2724 4243
701138219H1 SOYMON038 g1814402 BLASTN 594 1e-62 86 2725 4243
700965160H1 SOYMON022 g1814402 BLASTN 852 1e-62 91 2726 4243
701015168H1 SOYMON019 g1814402 BLASTN 855 1e-62 89 2727 4243
701136095H1 SOYMON038 g1814402 BLASTN 839 1e-61 88 2728 4243
700761789H1 SOYMON015 g1814402 BLASTN 845 1e-61 86 2729 4243
701105695H1 SOYMON036 g1814402 BLASTN 835 1e-60 87 2730 4243
700991714H1 SOYMON011 g1814402 BLASTN 502 1e-59 83 2731 4243
700564223H1 SOYMON002 g1814402 BLASTN 557 1e-59 90 2732 4243
700987384H1 SOYMON009 g1814402 BLASTN 803 1e-58 86 2733 4243
700833934H1 SOYMON019 g1814402 BLASTN 806 1e-58 89 2734 4243
700835181H1 SOYMON019 g1814402 BLASTN 806 1e-58 82 2735 4243
700737529H1 SOYMON010 g1814402 BLASTN 810 1e-58 92 2736 4243
701012851H1 SOYMON019 g1814402 BLASTN 811 1e-58 91 2737 4243
700556592H1 SOYMON001 g1814402 BLASTN 814 1e-58 88 2738 4243
700907579H1 SOYMON022 g1814402 BLASTN 781 1e-56 89 2739 4243
700961749H1 SOYMON022 g1814402 BLASTN 785 1e-56 91 2740 4243
700835239H1 SOYMON019 g1814402 BLASTN 787 1e-56 86 2741 4243
700646425H1 SOYMON013 g1814402 BLASTN 772 1e-55 89 2742 4243
701123924H1 SOYMON037 g1814402 BLASTN 775 1e-55 91 2743 4243
700957759H1 SOYMON022 g1814402 BLASTN 776 1e-55 90 2744 4243
700964425H1 SOYMON022 g1814402 BLASTN 777 1e-55 86 2745 4243
700962173H1 SOYMON022 g1814402 BLASTN 778 1e-55 91 2746 4243
701066293H1 SOYMON034 g1814402 BLASTN 759 1e-54 81 2747 4243
700986741H1 SOYMON009 g1814402 BLASTN 589 1e-53 84 2748 4243
701212514H1 SOYMON035 g1814402 BLASTN 591 1e-53 87 2749 4243
701009195H1 SOYMON019 g1814402 BLASTN 747 1e-53 91 2750 4243
701060510H1 SOYMON033 g1814402 BLASTN 748 1e-53 84 2751 4243
700848730H1 SOYMON021 g1814402 BLASTN 749 1e-53 90 2752 4243
700754870H1 SOYMON014 g1814402 BLASTN 751 1e-53 84 2753 4243
701212482H1 SOYMON035 g1814402 BLASTN 751 1e-53 84 2754 4243
700753283H1 SOYMON014 g1814402 BLASTN 752 1e-53 86 2755 4243
700738517H1 SOYMON012 g1814402 BLASTN 636 1e-52 89 2756 4243
700833978H1 SOYMON019 g1814402 BLASTN 740 1e-52 91 2757 4243
700756424H1 SOYMON014 g1814402 BLASTN 729 1e-51 82 2758 4243
701011759H1 SOYMON019 g1814402 BLASTN 729 1e-51 91 2759 4243
701010103H2 SOYMON019 g1814402 BLASTN 707 1e-50 82 2760 4243
700741392H1 SOYMON012 g1814402 BLASTN 707 1e-50 84 2761 4243
701123062H1 SOYMON037 g1814402 BLASTN 308 1e-49 88 2762 4243
701048949H1 SOYMON032 g1814402 BLASTN 502 1e-49 85 2763 4243
700834566H1 SOYMON019 g1814402 BLASTN 618 1e-49 88 2764 4243
700963965H1 SOYMON022 g1814402 BLASTN 685 1e-48 78 2765 4243
700986376H1 SOYMON009 g1814402 BLASTN 694 1e-48 84 2766 4243
701012708H1 SOYMON019 g1814402 BLASTN 521 1e-47 91 2767 4243
700746927H1 SOYMON013 g1814402 BLASTN 547 1e-46 78 2768 4243
700997448H1 SOYMON018 g1814402 BLASTN 470 1e-45 89 2769 4243
700830667H1 SOYMON019 g1814402 BLASTN 647 1e-45 86 2770 4243
700891479H1 SOYMON024 g1814402 BLASTN 654 1e-45 88 2771 4243
700562246H1 SOYMON002 g1814402 BLASTN 656 1e-45 86 2772 4243
701097325H1 SOYMON028 g1814402 BLASTN 420 1e-44 92 2773 4243
700835830H1 SOYMON019 g1814402 BLASTN 503 1e-44 86 2774 4243
700962969H1 SOYMON022 g1814402 BLASTN 515 1e-44 89 2775 4243
701102860H1 SOYMON028 g1814402 BLASTN 309 1e-42 92 2776 4243
700763506H1 SOYMON015 g1814402 BLASTN 456 1e-42 84
2777 4243 700994059H1 SOYMON011 g1814402 BLASTN 487 1e-42 87 2778
4243 700751551H1 SOYMON014 g1814402 BLASTN 568 1e-41 85 2779 4243
701108872H1 SOYMON036 g1814402 BLASTN 603 1e-41 85 2780 4243
700994053H1 SOYMON011 g1814402 BLASTN 587 1e-40 85 2781 4243
700729064H1 SOYMON009 g1814402 BLASTN 413 1e-38 90 2782 4243
700874176H1 SOYMON018 g1814402 BLASTN 468 1e-38 92 2783 4243
700742963H1 SOYMON012 g1814402 BLASTN 565 1e-38 86 2784 4243
701212923H1 SOYMON035 g1814402 BLASTN 572 1e-38 84 2785 4243
700994851H1 SOYMON011 g1814402 BLASTN 548 1e-36 76 2786 4243
701000193H1 SOYMON018 g1814402 BLASTN 296 1e-33 87 2787 4243
700756219H1 SOYMON014 g1814402 BLASTN 394 1e-32 80 2788 4243
701014751H1 SOYMON019 g1814402 BLASTN 461 1e-29 90 2789 4243
700561163H1 SOYMON002 g1814402 BLASTN 231 1e-25 89 2790 4243
700650284H1 SOYMON003 g974782 BLASTX 157 1e-14 96 2791 4243
700869218H1 SOYMON016 g1814402 BLASTN 248 1e-11 93 2792 550
LIB3028-007-Q1-B1-B6 LIB3028 g886470 BLASTN 1440 1e-111 81 2793 550
LIB3040-017-Q1-E1-E8 LIB3040 g886470 BLASTN 1260 1e-96 80 2794 550
700650656H1 SOYMON003 g1814402 BLASTN 1072 1e-93 83 2795 550
LIB3051-091-Q1-K1-C2 LIB3051 g886470 BLASTN 1124 1e-89 80 2796 550
LIB3051-072-Q1-K1-B3 LIB3051 g974781 BLASTN 1114 1e-83 82 2797 550
LIB3051-006-Q1-E1-G9 LIB3051 g974781 BLASTN 703 1e-82 81 2798 550
700563811H1 SOYMON002 g1814402 BLASTN 1071 1e-80 84 2799 550
700754104H1 SOYMON014 g974781 BLASTN 1052 1e-78 86 2800 550
701002973H1 SOYMON019 g1814402 BLASTN 1039 1e-77 85 2801 550
700986733H1 SOYMON009 g974781 BLASTN 1039 1e-77 83 2802 550
700557595H1 SOYMON001 g886470 BLASTN 1019 1e-76 83 2803 550
700563477H1 SOYMON002 g2738247 BLASTN 1024 1e-76 85 2804 550
700976112H1 SOYMON009 g886470 BLASTN 1024 1e-76 84 2805 550
701004484H1 SOYMON019 g1814402 BLASTN 1007 1e-75 84 2806 550
700889161H1 SOYMON024 g974781 BLASTN 996 1e-74 85 2807 550
700833110H1 SOYMON019 g974781 BLASTN 998 1e-74 87 2808 550
701124559H1 SOYMON037 g974781 BLASTN 1002 1e-74 86 2809 550
700729207H1 SOYMON009 g974781 BLASTN 1004 1e-74 85 2810 550
700730012H1 SOYMON009 g886470 BLASTN 987 1e-73 85 2811 550
700987128H1 SOYMON009 g2738247 BLASTN 989 1e-73 85 2812 550
700564788H1 SOYMON002 g974781 BLASTN 990 1e-73 84 2813 550
701099104H1 SOYMON028 g1814402 BLASTN 994 1e-73 86 2814 550
LIB3065-008-Q1-N1-B4 LIB3065 g2738247 BLASTN 839 1e-72 83 2815 550
701104283H1 SOYMON036 g974781 BLASTN 976 1e-72 83 2816 550
700726387H1 SOYMON009 g974781 BLASTN 980 1e-72 84 2817 550
700900884H1 SOYMON027 g1814402 BLASTN 982 1e-72 85 2818 550
LIB3051-029-Q1-K1-D6 LIB3051 g886470 BLASTN 824 1e-71 81 2819 550
701213995H1 SOYMON035 g1814402 BLASTN 962 1e-71 84 2820 550
700683596H1 SOYMON008 g1814402 BLASTN 968 1e-71 85 2821 550
700646529H1 SOYMON014 g1814402 BLASTN 867 1e-70 84 2822 550
700756704H1 SOYMON014 g1814402 BLASTN 893 1e-70 87 2823 550
700991647H1 SOYMON011 g1814402 BLASTN 947 1e-70 84 2824 550
700962195H1 SOYMON022 g974781 BLASTN 947 1e-70 87 2825 550
700994369H1 SOYMON011 g1814402 BLASTN 949 1e-70 84 2826 550
700672477H1 SOYMON006 g1814402 BLASTN 951 1e-70 85 2827 550
700946215H1 SOYMON024 g2738247 BLASTN 957 1e-70 83 2828 550
701152765H1 SOYMON031 g974781 BLASTN 958 1e-70 88 2829 550
701010552H1 SOYMON019 g974781 BLASTN 501 1e-69 83 2830 550
LIB3056-004-Q1-N1-B12 LIB3056 g2738247 BLASTN 698 1e-69 77 2831 550
701100656H1 SOYMON028 g974781 BLASTN 890 1e-69 85 2832 550
700985362H1 SOYMON009 g1814402 BLASTN 937 1e-69 82 2833 550
700746178H1 SOYMON013 g974781 BLASTN 938 1e-69 85 2834 550
700674440H1 SOYMON007 g974781 BLASTN 938 1e-69 83 2835 550
700556934H1 SOYMON001 g1814402 BLASTN 938 1e-69 82 2836 550
700652624H1 SOYMON003 g1814402 BLASTN 939 1e-69 82 2837 550
700952331H1 SOYMON022 g886470 BLASTN 946 1e-69 83 2838 550
700725576H1 SOYMON009 g886470 BLASTN 946 1e-69 85 2839 550
701003602H1 SOYMON019 g2738247 BLASTN 924 1e-68 83 2840 550
700745053H1 SOYMON013 g974781 BLASTN 924 1e-68 85 2841 550
700895781H1 SOYMON027 g1814402 BLASTN 926 1e-68 84 2842 550
700664593H1 SOYMON005 g974781 BLASTN 926 1e-68 87 2843 550
700864264H1 SOYMON016 g1814402 BLASTN 930 1e-68 84 2844 550
700674466H1 SOYMON007 g974781 BLASTN 933 1e-68 83 2845 550
700564170H1 SOYMON002 g974781 BLASTN 861 1e-67 83 2846 550
700654531H1 SOYMON004 g974781 BLASTN 911 1e-67 81 2847 550
700996341H1 SOYMON018 g886470 BLASTN 911 1e-67 83 2848 550
701136257H1 SOYMON038 g886470 BLASTN 912 1e-67 82 2849 550
700751114H1 SOYMON014 g1814402 BLASTN 914 1e-67 84 2850 550
700657606H1 SOYMON004 g1814402 BLASTN 916 1e-67 87 2851 550
700983827H1 SOYMON009 g974781 BLASTN 918 1e-67 85 2852 550
700981249H1 SOYMON009 g1814402 BLASTN 921 1e-67 80 2853 550
700836103H1 SOYMON019 g886470 BLASTN 922 1e-67 82 2854 550
701011869H1 SOYMON019 g886470 BLASTN 486 1e-66 86 2855 550
701097045H1 SOYMON028 g886470 BLASTN 601 1e-66 85 2856 550
701012695H1 SOYMON019 g974781 BLASTN 777 1e-66 85 2857 550
700945396H1 SOYMON024 g974781 BLASTN 902 1e-66 85 2858 550
700755390H1 SOYMON014 g974781 BLASTN 903 1e-66 86 2859 550
700967721H1 SOYMON033 g886470 BLASTN 910 1e-66 82 2860 550
700750610H1 SOYMON014 g974781 BLASTN 910 1e-66 84 2861 550
700908231H1 SOYMON022 g974781 BLASTN 527 1e-65 83 2862 550
701003835H1 SOYMON019 g886470 BLASTN 723 1e-65 82 2863 550
700790802H1 SOYMON011 g2738247 BLASTN 891 1e-65 82 2864 550
701053438H1 SOYMON032 g974781 BLASTN 893 1e-65 82 2865 550
701009430H1 SOYMON019 g974781 BLASTN 894 1e-65 85 2866 550
700891415H1 SOYMON024 g1814402 BLASTN 895 1e-65 84 2867 550
701100681H1 SOYMON028 g2738247 BLASTN 482 1e-64 84 2868 550
700893420H1 SOYMON024 g974781 BLASTN 570 1e-64 89 2869 550
700752741H1 SOYMON014 g886470 BLASTN 880 1e-64 82 2870 550
700955938H1 SOYMON022 g1814402 BLASTN 882 1e-64 81 2871 550
701015042H1 SOYMON019 g2738247 BLASTN 886 1e-64 82 2872 550
LIB3051-072-Q1-K1-B1 LIB3051 g974781 BLASTN 645 1e-63 76 2873 550
700741918H1 SOYMON012 g886470 BLASTN 722 1e-63 84 2874 550
701103319H1 SOYMON028 g974781 BLASTN 792 1e-63 82 2875 550
700833218H1 SOYMON019 g2738247 BLASTN 863 1e-63 81 2876 550
701008071H1 SOYMON019 g886470 BLASTN 867 1e-63 83 2877 550
700832073H1 SOYMON019 g974781 BLASTN 871 1e-63 83 2878 550
700889695H1 SOYMON024 g974781 BLASTN 872 1e-63 83 2879 550
701007489H2 SOYMON019 g974781 BLASTN 873 1e-63 84 2880 550
700895858H1 SOYMON027 g974781 BLASTN 874 1e-63 84 2881 550
700753955H1 SOYMON014 g974781 BLASTN 470 1e-62 87 2882 550
700564433H1 SOYMON002 g886470 BLASTN 757 1e-62 84 2883 550
700963115H1 SOYMON022 g974781 BLASTN 851 1e-62 83 2884 550
700894728H1 SOYMON024 g886470 BLASTN 860 1e-62 84 2885 550
701056915H1 SOYMON033 g886470 BLASTN 862 1e-62 84 2886 550
700741134H1 SOYMON012 g886470 BLASTN 862 1e-62 83 2887 550
700847591H1 SOYMON021 g974781 BLASTN 842 1e-61 84 2888 550
700941253H1 SOYMON024 g2738247 BLASTN 844 1e-61 80 2889 550
701004315H1 SOYMON019 g2738247 BLASTN 846 1e-61 83 2890 550
700895720H1 SOYMON027 g974781 BLASTN 848 1e-61 83 2891 550
701013541H1 SOYMON019 g974781 BLASTN 849 1e-61 84 2892 550
700892552H1 SOYMON024 g886470 BLASTN 719 1e-60 83 2893 550
701141313H1 SOYMON038 g974781 BLASTN 827 1e-60 83 2894 550
701012547H1 SOYMON019 g1814402 BLASTN 831 1e-60 83 2895 550
701008558H1 SOYMON019 g1814402 BLASTN 831 1e-60 83 2896 550
700902022H1 SOYMON027 g2738247 BLASTN 831 1e-60 82 2897 550
700959515H1 SOYMON022 g886470 BLASTN 832 1e-60 81 2898 550
701042630H1 SOYMON029 g1814402 BLASTN 819 1e-59 81 2899 550
700941292H1 SOYMON024 g2738247 BLASTN 820 1e-59 81 2900 550
700788526H1 SOYMON011 g974781 BLASTN 491 1e-58 82 2901 550
700894839H1 SOYMON024 g1814402 BLASTN 498 1e-58 82 2902 550
700865873H1 SOYMON016 g974781 BLASTN 808 1e-58 85 2903 550
701015435H1 SOYMON019 g886470 BLASTN 809 1e-58 80 2904 550
700755960H1 SOYMON014 g2738247 BLASTN 809 1e-58 78 2905 550
700876051H1 SOYMON018 g886470 BLASTN 434 1e-57 81 2906 550
701041327H1 SOYMON029 g886470 BLASTN 499 1e-57 83 2907 550
701098902H1 SOYMON028 g2738247 BLASTN 767 1e-57 77 2908 550
700853392H1 SOYMON023 g886470 BLASTN 793 1e-57 82 2909 550
700872645H1 SOYMON018 g974781 BLASTN 798 1e-57 83 2910 550
700989675H1 SOYMON011 g2738247 BLASTN 800 1e-57 79 2911 550
700753487H1 SOYMON014 g1814402 BLASTN 589 1e-56 83 2912 550
700736276H1 SOYMON010 g1814402 BLASTN 782 1e-56 79 2913 550
700891361H1 SOYMON024 g1814402 BLASTN 783 1e-56 81 2914 550
700829712H1 SOYMON019 g974781 BLASTN 790 1e-56 80 2915 550
LIB3050-019-Q1-K1-A1 LIB3050 g974781 BLASTN 663 1e-55 81 2916 550
701001013H1 SOYMON018 g1814402 BLASTN 768 1e-55 84 2917 550
LIB3028-031-Q1-B1-G12 LIB3028 g886470 BLASTN 775 1e-55 82 2918 550
701212782H1 SOYMON035 g886470 BLASTN 621 1e-54 82 2919 550
701008695H1 SOYMON019 g974781 BLASTN 718 1e-54 80 2920 550
700990972H1 SOYMON011 g1814402 BLASTN 766 1e-54 81 2921 550
700789576H2 SOYMON011 g886470 BLASTN 766 1e-54 80 2922 550
700994266H1 SOYMON011 g1814402 BLASTN 408 1e-53 85 2923 550
700731985H1 SOYMON010 g974781 BLASTN 590 1e-53 81 2924 550
700907927H1 SOYMON022 g886470 BLASTN 612 1e-53 80 2925 550
701012079H1 SOYMON019 g974781 BLASTN 669 1e-53 86 2926 550
700753939H1 SOYMON014 g886470 BLASTN 690 1e-53 78 2927 550
701000754H1 SOYMON018 g974781 BLASTN 745 1e-53 76 2928 550
701040287H1 SOYMON029 g886470 BLASTN 747 1e-53 82 2929 550
700891329H1 SOYMON024 g974781 BLASTN 749 1e-53 79 2930 550
700897258H1 SOYMON027 g886470 BLASTN 752 1e-53 86 2931 550
700944949H1 SOYMON024 g974781 BLASTN 426 1e-52 82 2932 550
701108671H1 SOYMON036 g1814402 BLASTN 731 1e-52 77 2933 550
700905233H1 SOYMON022 g886470 BLASTN 732 1e-52 77 2934 550
700958589H1 SOYMON022 g886470 BLASTN 738 1e-52 80 2935 550
700666436H1 SOYMON005 g2738247 BLASTN 739 1e-52 82 2936 550
700829902H1 SOYMON019 g886470 BLASTN 740 1e-52 82 2937 550
700740110H1 SOYMON012 g974781 BLASTN 742 1e-52 83 2938 550
700989055H1 SOYMON011 g886470 BLASTN 538 1e-50 79 2939 550
701213370H1 SOYMON035 g974781 BLASTN 599 1e-50 83 2940 550
701098072H1 SOYMON028 g974781 BLASTN 709 1e-50 74 2941 550
700896128H1 SOYMON027 g886470 BLASTN 714 1e-50 83 2942 550
701060755H1 SOYMON033 g974781 BLASTN 716 1e-50 74 2943 550
700953594H1 SOYMON022 g1814402 BLASTN 695 1e-49 78 2944 550
701046911H1 SOYMON032 g2738247 BLASTN 687 1e-48 81 2945 550
701065707H1 SOYMON034 g1814402 BLASTN 443 1e-47 84 2946 550
700962114H1 SOYMON022 g886470 BLASTN 678 1e-47 84 2947 550
700831826H1 SOYMON019 g2738247 BLASTN 682 1e-47 77 2948 550
701054296H1 SOYMON032 g886470 BLASTN 454 1e-46 83 2949 550
700888738H1 SOYMON024 g974781 BLASTN 552 1e-46 77 2950 550
700892022H1 SOYMON024 g886470 BLASTN 605 1e-46 81 2951 550
700890275H1 SOYMON024 g2738247 BLASTN 666 1e-46 77 2952 550
LIB3051-006-Q1-K1-G9 LIB3051 g974781 BLASTN 670 1e-45 80 2953 550
700889113H1 SOYMON024 g886470 BLASTN 582 1e-43 81 2954 550
700952720H1 SOYMON022 g886470 BLASTN 611 1e-42 76 2955 550
701014761H1 SOYMON019 g886470 BLASTN 318 1e-41 84 2956 550
700753882H1 SOYMON014 g886470 BLASTN 381 1e-41 79 2957 550
700743792H1 SOYMON012 g1814402 BLASTN 610 1e-41 86 2958 550
700990963H1 SOYMON011 g886470 BLASTN 578 1e-39 77 2959 550
700941880H1 SOYMON024 g2738247 BLASTN 583 1e-39 81 2960 550
700898962H1 SOYMON027 g2738247 BLASTN 571 1e-38 80 2961 550
700990865H1 SOYMON011 g2738247 BLASTN 467 1e-37 77 2962 550
700565779H1 SOYMON002 g886470 BLASTN 557 1e-37 70 2963 550
700993903H1 SOYMON011 g974781 BLASTN 562 1e-37 84 2964 550
700941589H1 SOYMON024 g2738247 BLASTN 550 1e-36 81 2965 550
701052554H1 SOYMON032 g886470 BLASTN 534 1e-35 67 2966 550
700991055H1 SOYMON011 g886470 BLASTN 356 1e-34 79 2967 550
701010438H1 SOYMON019 g2738247 BLASTN 508 1e-33 79 2968 550
700756634H1 SOYMON014 g2738247 BLASTN 494 1e-32 76 2969 550
701042980H1 SOYMON029 g886470 BLASTN 461 1e-29 83 2970 550
700682940H1 SOYMON008 g2738247 BLASTN 466 1e-29 83 2971 550
701049575H1 SOYMON032 g886470 BLASTN 433 1e-27 82 2972 550
700982552H1 SOYMON009 g886470 BLASTN 356 1e-25 78 2973 550
700675637H1 SOYMON007 g886470 BLASTN 375 1e-25 79 2974 550
701142153H1 SOYMON038 g886470 BLASTN 377 1e-22 76 2975 550
700682724H1 SOYMON008 g974781 BLASTN 361 1e-19 88 2976 550
701051764H1 SOYMON032 g1814402 BLASTN 211 1e-17 80 2977 550
700867241H1 SOYMON016 g2738248 BLASTX 152 1e-13 88 2978 550
701054954H1 SOYMON032 g2738248 BLASTX 138 1e-12 86 2979 550
700790450H2 SOYMON011 g974781 BLASTN 238 1e-10 80 2980 550
700653979H1 SOYMON003 g1814403 BLASTX 118 1e-9 92 2981 550
700894218H1 SOYMON024 g2738248 BLASTX 122 1e-9 78 2982 550
700863078H1 SOYMON022 g2738247 BLASTN 236 1e-8 78 2983 5758
701209304H1 SOYMON035 g886470 BLASTN 766 1e-54 84 2984 5758
701106455H1 SOYMON036 g886470 BLASTN 723 1e-51 82 2985 5758
700833538H1 SOYMON019 g1814402 BLASTN 609 1e-43 83 2986 5758
701051425H1 SOYMON032 g886470 BLASTN 629 1e-43 83 2987 5758
700654506H1 SOYMON004 g886470 BLASTN 438 1e-26 70 2988 5758
701047795H1 SOYMON032 g974782 BLASTX 161 1e-15 100 2989 5758
701202409H1 SOYMON035 g1814402 BLASTN 310 1e-15 79 2990 8266
700558628H1 SOYMON001 g886470 BLASTN 780 1e-56 74 2991 8266
701207720H1 SOYMON035 g1814402 BLASTN 766 1e-54 74 2992 8266
700557429H1 SOYMON001 g1814402 BLASTN 728 1e-51 75
ADENOSYLHOMOCYSTEINASE (EC 3.3.1.1) 1354 -700154280 700154280H1
SATMON007 g170772 BLASTN 229 1e-23 77 1355 -L30594291
LIB3059-032-Q1-K1-A8 LIB3059 g170772 BLASTN 516 1e-74 73 1356
-L30664307 LIB3066-047-Q1-K1-E7 LIB3066 g170772 BLASTN 628 1e-49 70
1357 503 LIB3079-013-Q1-K1-D3 LIB3079 g170772 BLASTN 1505 1e-139 89
1358 503 LIB3062-017-Q1-K1-A10 LIB3062 g170772 BLASTN 1571 1e-129
89 1359 503 LIB3067-001-Q1-K1-D11 LIB3067 g170772 BLASTN 1654
1e-129 89 1360 503 LIB148-060-Q1-E1-B9 LIB148 g170772 BLASTN 1620
1e-126 90 1361 503 LIB3069-043-Q1-K1-G4 LIB3069 g170772 BLASTN 1360
1e-124 84 1362 503 LIB143-006-Q1-E1-F3 LIB143 g170772 BLASTN 831
1e-120 88 1363 503 LIB189-009-Q1-E1-E9 LIB189 g170772 BLASTN 1551
1e-120 90 1364 503 LIB3060-003-Q1-K1-F9 LIB3060 g170772 BLASTN 1538
1e-119 91 1365 503 700089023H1 SATMON011 g170772 BLASTN 1495 1e-115
91 1366 503 LIB143-061-Q1-E1-C9 LIB143 g170772 BLASTN 1474 1e-114
90 1367 503 LIB3069-034-Q1-K1-E5 LIB3069 g170772 BLASTN 1483 1e-114
88 1368 503 LIB143-061-Q1-E1-E5 LIB143 g170772 BLASTN 1440 1e-113
89 1369 503 LIB3067-040-Q1-K1-H5 LIB3067 g170772 BLASTN 1467 1e-113
88 1370 503 LIB3067-048-Q1-K1-A11 LIB3067 g170772 BLASTN 1207
1e-109 86 1371 503 LIB3068-050-Q1-K1-G6 LIB3068 g170772 BLASTN 1279
1e-109 86 1372 503 LIB3069-036-Q1-K1-F6 LIB3069 g170772 BLASTN 1310
1e-109 92 1373 503 LIB3066-047-Q1-K1-H5 LIB3066 g170772 BLASTN 1353
1e-108 86 1374 503 LIB3059-011-Q1-K1-A5 LIB3059 g170772 BLASTN 1401
1e-107 90 1375 503 LIB3067-056-Q1-K1-E11 LIB3067 g170772 BLASTN 957
1e-106 86 1376 503 LIB189-010-Q1-E1-E12 LIB189 g170772 BLASTN 1144
1e-106 88 1377 503 700084426H1 SATMON011 g170772 BLASTN 1368 1e-105
91 1378 503 700086273H1 SATMON011 g170772 BLASTN 1364 1e-104 91
1379 503 700573027H1 SATMON030 g170772 BLASTN 939 1e-103 89 1380
503 700209360H1 SATMON016 g170772 BLASTN 1350 1e-103 89 1381 503
700619916H1 SATMON034 g170772 BLASTN 1050 1e-102 89 1382 503
700086051H1 SATMON011 g170772 BLASTN 1336 1e-102 90 1383 503
LIB3069-034-Q1-K1-C8 LIB3069 g170772 BLASTN 1322 1e-101 86 1384 503
700026324H1 SATMON003 g170772 BLASTN 1328 1e-101 91 1385 503
700104549H1 SATMON010 g170772 BLASTN 836 1e-100 88 1386 503
700622108H1 SATMON034 g170772 BLASTN 1158 1e-100 88 1387 503
700093980H1 SATMON008 g170772 BLASTN 1312 1e-100 90
1388 503 700077427H1 SATMON007 g170772 BLASTN 1317 1e-100 90 1389
503 700095389H1 SATMON008 g170772 BLASTN 1302 1e-99 91 1390 503
LIB3067-055-Q1-K1-D3 LIB3067 g170772 BLASTN 762 1e-98 87 1391 503
LIB3060-054-Q1-K1-F6 LIB3060 g170772 BLASTN 1009 1e-98 83 1392 503
700083339H1 SATMON011 g170772 BLASTN 1282 1e-98 91 1393 503
700102631H1 SATMON010 g170772 BLASTN 1283 1e-98 89 1394 503
700265625H1 SATMON017 g170772 BLASTN 1286 1e-98 90 1395 503
700095002H1 SATMON008 g170772 BLASTN 1289 1e-98 88 1396 503
700094761H1 SATMON008 g170772 BLASTN 1289 1e-98 88 1397 503
700073832H1 SATMON007 g170772 BLASTN 1270 1e-97 88 1398 503
700047817H1 SATMON003 g170772 BLASTN 1272 1e-97 91 1399 503
700091149H1 SATMON011 g170772 BLASTN 1262 1e-96 89 1400 503
700098584H1 SATMON009 g170772 BLASTN 1264 1e-96 91 1401 503
700085932H1 SATMON011 g170772 BLASTN 1266 1e-96 89 1402 503
700094081H1 SATMON008 g170772 BLASTN 1269 1e-96 91 1403 503
700202442H1 SATMON003 g170772 BLASTN 1145 1e-95 88 1404 503
700049770H1 SATMON003 g170772 BLASTN 1246 1e-95 89 1405 503
LIB3079-020-Q1-K1-B12 LIB3079 g170772 BLASTN 1249 1e-95 85 1406 503
700090226H1 SATMON011 g170772 BLASTN 1252 1e-95 90 1407 503
700088191H1 SATMON011 g170772 BLASTN 1253 1e-95 90 1408 503
700076743H1 SATMON007 g170772 BLASTN 1256 1e-95 90 1409 503
LIB189-009-Q1-E1-E10 LIB189 g170772 BLASTN 789 1e-94 88 1410 503
700072038H1 SATMON007 g170772 BLASTN 1237 1e-94 89 1411 503
700082967H1 SATMON011 g170772 BLASTN 1239 1e-94 89 1412 503
LIB3068-062-Q1-K1-A3 LIB3068 g170772 BLASTN 1148 1e-93 82 1413 503
700094713H1 SATMON008 g170772 BLASTN 1222 1e-93 88 1414 503
700095620H1 SATMON008 g170772 BLASTN 1173 1e-92 89 1415 503
700242509H1 SATMON010 g170772 BLASTN 1213 1e-92 92 1416 503
700071923H1 SATMON007 g170772 BLASTN 1214 1e-92 90 1417 503
700575314H1 SATMON030 g170772 BLASTN 1149 1e-91 88 1418 503
700086654H1 SATMON011 g170772 BLASTN 1199 1e-91 90 1419 503
700241072H1 SATMON010 g170772 BLASTN 1205 1e-91 91 1420 503
700047361H1 SATMON003 g170772 BLASTN 1110 1e-90 89 1421 503
700217056H1 SATMON016 g170772 BLASTN 1187 1e-90 91 1422 503
LIB3067-005-Q1-K1-F2 LIB3067 g170772 BLASTN 1188 1e-90 84 1423 503
700075495H1 SATMON007 g170772 BLASTN 1193 1e-90 86 1424 503
700084783H1 SATMON011 g170772 BLASTN 1194 1e-90 91 1425 503
LIB143-059-Q1-E1-C3 LIB143 g170772 BLASTN 952 1e-89 84 1426 503
700448818H1 SATMON028 g170772 BLASTN 1174 1e-89 91 1427 503
700159079H1 SATMON012 g170772 BLASTN 1177 1e-89 92 1428 503
700094408H1 SATMON008 g170772 BLASTN 1177 1e-89 92 1429 503
700348440H1 SATMON023 g170772 BLASTN 1182 1e-89 89 1430 503
700077082H1 SATMON007 g170772 BLASTN 1182 1e-89 92 1431 503
700082111H1 SATMON011 g170772 BLASTN 1183 1e-89 89 1432 503
700239752H1 SATMON010 g170772 BLASTN 1164 1e-88 89 1433 503
700029456H1 SATMON003 g170772 BLASTN 1166 1e-88 90 1434 503
700209430H1 SATMON016 g170772 BLASTN 1170 1e-88 90 1435 503
700208660H1 SATMON016 g170772 BLASTN 930 1e-87 90 1436 503
700213138H1 SATMON016 g170772 BLASTN 1042 1e-87 89 1437 503
700102234H1 SATMON010 g170772 BLASTN 634 1e-85 90 1438 503
700450479H1 SATMON028 g170772 BLASTN 1015 1e-85 90 1439 503
700095372H1 SATMON008 g170772 BLASTN 1126 1e-85 90 1440 503
700218734H1 SATMON011 g170772 BLASTN 1128 1e-85 94 1441 503
700256858H1 SATMON017 g170772 BLASTN 1133 1e-85 90 1442 503
700221063H1 SATMON011 g170772 BLASTN 1137 1e-85 90 1443 503
700105439H1 SATMON010 g170772 BLASTN 1118 1e-84 89 1444 503
700085174H1 SATMON011 g170772 BLASTN 1125 1e-84 90 1445 503
700242421H1 SATMON010 g170772 BLASTN 1125 1e-84 92 1446 503
700077492H1 SATMON007 g170772 BLASTN 774 1e-83 88 1447 503
700209295H1 SATMON016 g170772 BLASTN 914 1e-83 89 1448 503
700455826H1 SATMON029 g170772 BLASTN 978 1e-83 91 1449 503
700213370H1 SATMON016 g170772 BLASTN 1110 1e-83 92 1450 503
700352095H1 SATMON023 g170772 BLASTN 1111 1e-83 89 1451 503
700076842H1 SATMON007 g170772 BLASTN 1112 1e-83 90 1452 503
700215872H1 SATMON016 g170772 BLASTN 1113 1e-83 89 1453 503
700073645H1 SATMON007 g170772 BLASTN 1113 1e-83 89 1454 503
700048153H1 SATMON003 g170772 BLASTN 651 1e-82 90 1455 503
700073307H1 SATMON007 g170772 BLASTN 762 1e-82 90 1456 503
700350009H1 SATMON023 g170772 BLASTN 924 1e-82 88 1457 503
LIB143-059-Q1-E1-C5 LIB143 g170772 BLASTN 1029 1e-82 83 1458 503
700073955H1 SATMON007 g170772 BLASTN 1090 1e-82 90 1459 503
LIB3059-042-Q1-K1-H12 LIB3059 g170772 BLASTN 1090 1e-82 85 1460 503
700238024H1 SATMON010 g170772 BLASTN 1091 1e-82 90 1461 503
700155863H1 SATMON007 g170772 BLASTN 1091 1e-82 93 1462 503
700217890H1 SATMON016 g170772 BLASTN 1093 1e-82 91 1463 503
700239314H1 SATMON010 g170772 BLASTN 1094 1e-82 88 1464 503
700048337H1 SATMON003 g170772 BLASTN 1094 1e-82 92 1465 503
700235469H1 SATMON010 g170772 BLASTN 1098 1e-82 91 1466 503
700164224H1 SATMON013 g170772 BLASTN 710 1e-81 92 1467 503
700209374H1 SATMON016 g170772 BLASTN 1083 1e-81 89 1468 503
700082268H1 SATMON011 g170772 BLASTN 1083 1e-81 88 1469 503
700159326H1 SATMON012 g170772 BLASTN 777 1e-80 91 1470 503
700025934H1 SATMON003 g170772 BLASTN 792 1e-80 91 1471 503
700210458H1 SATMON016 g170772 BLASTN 856 1e-80 88 1472 503
700243826H1 SATMON010 g170772 BLASTN 881 1e-80 90 1473 503
700242676H1 SATMON010 g170772 BLASTN 940 1e-80 89 1474 503
700345565H1 SATMON021 g170772 BLASTN 1034 1e-80 89 1475 503
700264733H1 SATMON017 g170772 BLASTN 1044 1e-80 90 1476 503
700159161H1 SATMON012 g170772 BLASTN 1068 1e-80 90 1477 503
700072491H1 SATMON007 g170772 BLASTN 604 1e-79 88 1478 503
700053204H1 SATMON008 g170772 BLASTN 618 1e-79 89 1479 503
700451515H1 SATMON028 g170772 BLASTN 1057 1e-79 85 1480 503
700468929H1 SATMON025 g170772 BLASTN 1061 1e-79 84 1481 503
700205801H1 SATMON003 g170772 BLASTN 1064 1e-79 91 1482 503
700611326H1 SATMON022 g170772 BLASTN 725 1e-78 88 1483 503
700082291H1 SATMON011 g170772 BLASTN 1043 1e-78 87 1484 503
700071695H1 SATMON007 g170772 BLASTN 1048 1e-78 89 1485 503
700551561H1 SATMON022 g170772 BLASTN 746 1e-77 88 1486 503
700221019H1 SATMON011 g170772 BLASTN 856 1e-77 89 1487 503
LIB3078-007-Q1-K1-C5 LIB3078 g170772 BLASTN 936 1e-77 83 1488 503
700049925H1 SATMON003 g170772 BLASTN 954 1e-77 89 1489 503
700154101H1 SATMON007 g170772 BLASTN 1036 1e-77 88 1490 503
700380151H1 SATMON021 g170772 BLASTN 613 1e-76 88 1491 503
700243831H1 SATMON010 g170772 BLASTN 842 1e-76 89 1492 503
700235671H1 SATMON010 g170772 BLASTN 1019 1e-76 90 1493 503
700087847H1 SATMON011 g170772 BLASTN 1009 1e-75 82 1494 503
700208747H1 SATMON016 g170772 BLASTN 1010 1e-75 89 1495 503
700071795H1 SATMON007 g170772 BLASTN 1011 1e-75 89 1496 503
700157002H1 SATMON012 g170772 BLASTN 1012 1e-75 90 1497 503
700201608H1 SATMON003 g170772 BLASTN 1015 1e-75 85 1498 503
700451541H1 SATMON028 g170772 BLASTN 1015 1e-75 86 1499 503
700212182H1 SATMON016 g170772 BLASTN 1015 1e-75 88 1500 503
700381485H1 SATMON023 g170772 BLASTN 1017 1e-75 91 1501 503
700096793H1 SATMON008 g170772 BLASTN 761 1e-74 87 1502 503
700093025H1 SATMON008 g170772 BLASTN 996 1e-74 89 1503 503
700017129H1 SATMON001 g170772 BLASTN 998 1e-74 92 1504 503
700216525H1 SATMON016 g170772 BLASTN 1000 1e-74 86 1505 503
700087912H1 SATMON011 g170772 BLASTN 1001 1e-74 91 1506 503
700172618H1 SATMON013 g170772 BLASTN 1001 1e-74 91 1507 503
700801894H1 SATMON036 g170772 BLASTN 1002 1e-74 90 1508 503
700162040H1 SATMON012 g170772 BLASTN 1003 1e-74 92 1509 503
700212084H1 SATMON016 g170772 BLASTN 1004 1e-74 89 1510 503
700027971H1 SATMON003 g170772 BLASTN 982 1e-73 92 1511 503
700077321H1 SATMON007 g170772 BLASTN 985 1e-73 89 1512 503
700619884H1 SATMON034 g170772 BLASTN 987 1e-73 87 1513 503
700457201H1 SATMON029 g170772 BLASTN 988 1e-73 87 1514 503
700082970H1 SATMON011 g170772 BLASTN 990 1e-73 89 1515 503
700083350H1 SATMON011 g170772 BLASTN 971 1e-72 89 1516 503
700017732H1 SATMON001 g170772 BLASTN 973 1e-72 88 1517 503
700094688H1 SATMON008 g170772 BLASTN 975 1e-72 89 1518 503
700170985H1 SATMON013 g170772 BLASTN 979 1e-72 89 1519 503
700451847H1 SATMON028 g170772 BLASTN 872 1e-71 84 1520 503
700455091H1 SATMON029 g170772 BLASTN 931 1e-71 93 1521 503
700106189H1 SATMON010 g170772 BLASTN 960 1e-71 88 1522 503
700160075H1 SATMON012 g170772 BLASTN 968 1e-71 90 1523 503
700084952H1 SATMON011 g170772 BLASTN 552 1e-70 78 1524 503
700348238H1 SATMON023 g170772 BLASTN 946 1e-70 87 1525 503
700151310H1 SATMON007 g170772 BLASTN 948 1e-70 90 1526 503
700048962H1 SATMON003 g170772 BLASTN 677 1e-69 88 1527 503
700088485H1 SATMON011 g170772 BLASTN 936 1e-69 88 1528 503
700093667H1 SATMON008 g170772 BLASTN 937 1e-69 88 1529 503
700161491H1 SATMON012 g170772 BLASTN 928 1e-68 86 1530 503
700073939H1 SATMON007 g170772 BLASTN 931 1e-68 88 1531 503
700027715H1 SATMON003 g170772 BLASTN 472 1e-67 90 1532 503
700072843H1 SATMON007 g170772 BLASTN 692 1e-67 87 1533 503
700439356H1 SATMON026 g170772 BLASTN 777 1e-67 85 1534 503
700073556H1 SATMON007 g170772 BLASTN 910 1e-67 88 1535 503
700154461H1 SATMON007 g170772 BLASTN 911 1e-67 87 1536 503
700201909H1 SATMON003 g170772 BLASTN 912 1e-67 88 1537 503
700104384H1 SATMON010 g170772 BLASTN 665 1e-66 89 1538 503
700152707H1 SATMON007 g170772 BLASTN 898 1e-66 88 1539 503
700076625H1 SATMON007 g170772 BLASTN 900 1e-66 85 1540 503
700456945H1 SATMON029 g170772 BLASTN 902 1e-66 80 1541 503
700457294H1 SATMON029 g170772 BLASTN 446 1e-65 88 1542 503
700152062H1 SATMON007 g170772 BLASTN 803 1e-65 90 1543 503
700072428H2 SATMON007 g170772 BLASTN 890 1e-65 88 1544 503
700217352H1 SATMON016 g170772 BLASTN 893 1e-65 88 1545 503
700072443H2 SATMON007 g170772 BLASTN 894 1e-65 89 1546 503
700087436H1 SATMON011 g170772 BLASTN 874 1e-64 88 1547 503
700240615H1 SATMON010 g170772 BLASTN 877 1e-64 88 1548 503
700155233H1 SATMON007 g170772 BLASTN 883 1e-64 90 1549 503
700241210H1 SATMON010 g170772 BLASTN 822 1e-63 87 1550 503
700478070H1 SATMON025 g170772 BLASTN 863 1e-63 82 1551 503
700447433H1 SATMON027 g170772 BLASTN 865 1e-63 84 1552 503
700215884H1 SATMON016 g170772 BLASTN 865 1e-63 88 1553 503
700575250H1 SATMON030 g170772 BLASTN 820 1e-61 84 1554 503
700209283H1 SATMON016 g170772 BLASTN 833 1e-60 86 1555 503
700213479H1 SATMON016 g170772 BLASTN 834 1e-60 88 1556 503
700241740H1 SATMON010 g170772 BLASTN 814 1e-59 88 1557 503
700026012H1 SATMON003 g170772 BLASTN 815 1e-59 87 1558 503
700151036H1 SATMON007 g170772 BLASTN 816 1e-59 91 1559 503
700207150H1 SATMON017 g170772 BLASTN 818 1e-59 87 1560 503
700217383H1 SATMON016 g170772 BLASTN 824 1e-59 88 1561 503
700618168H1 SATMON033 g170772 BLASTN 385 1e-58 80 1562 503
700020777H1 SATMON001 g170772 BLASTN 806 1e-58 85 1563 503
700224131H1 SATMON011 g170772 BLASTN 794 1e-57 88 1564 503
700222876H1 SATMON011 g170772 BLASTN 794 1e-57 88 1565 503
700027716H1 SATMON003 g170772 BLASTN 642 1e-56 79 1566 503
700073305H1 SATMON007 g170772 BLASTN 787 1e-56 83 1567 503
700257376H1 SATMON017 g170772 BLASTN 362 1e-55 87 1568 503
700151752H1 SATMON007 g170772 BLASTN 766 1e-55 89 1569 503
700219086H1 SATMON011 g170772 BLASTN 769 1e-55 87 1570 503
700218924H1 SATMON011 g170772 BLASTN 775 1e-55 87 1571 503
700424527H1 SATMONN01 g170772 BLASTN 777 1e-55 83 1572 503
700073183H1 SATMON007 g170772 BLASTN 607 1e-54 85 1573 503
700217412H1 SATMON016 g170772 BLASTN 755 1e-54 87 1574 503
700208634H1 SATMON016 g170772 BLASTN 642 1e-53 83 1575 503
700202577H1 SATMON003 g170772 BLASTN 743 1e-53 86 1576 503
700446645H1 SATMON027 g170772 BLASTN 744 1e-53 87 1577 503
700238061H1 SATMON010 g170772 BLASTN 745 1e-53 86 1578 503
700617325H1 SATMON033 g170772 BLASTN 751 1e-53 91 1579 503
700239901H1 SATMON010 g170772 BLASTN 641 1e-52 86 1580 503
700155130H1 SATMON007 g170772 BLASTN 646 1e-52 88 1581 503
700083360H1 SATMON011 g170772 BLASTN 739 1e-52 87 1582 503
700236915H1 SATMON010 g170772 BLASTN 740 1e-52 87 1583 503
700074780H1 SATMON007 g170772 BLASTN 528 1e-51 83 1584 503
700479515H1 SATMON034 g170772 BLASTN 718 1e-51 88 1585 503
700215545H1 SATMON016 g170772 BLASTN 724 1e-51 90 1586 503
700165354H1 SATMON013 g170772 BLASTN 728 1e-51 86 1587 503
700353994H1 SATMON024 g170772 BLASTN 713 1e-50 87 1588 503
700153619H1 SATMON007 g170772 BLASTN 694 1e-49 86 1589 503
700074779H1 SATMON007 g170772 BLASTN 705 1e-49 77 1590 503
700155530H1 SATMON007 g170772 BLASTN 686 1e-48 86 1591 503
700221207H1 SATMON011 g170772 BLASTN 692 1e-48 86 1592 503
700264257H1 SATMON017 g170772 BLASTN 672 1e-47 87 1593 503
700152216H1 SATMON007 g170772 BLASTN 659 1e-46 88 1594 503
700150643H1 SATMON007 g170772 BLASTN 659 1e-46 88 1595 503
700156027H1 SATMON007 g170772 BLASTN 655 1e-45 93 1596 503
700260335H1 SATMON017 g170772 BLASTN 385 1e-44 79 1597 503
700623795H1 SATMON034 g170772 BLASTN 401 1e-44 88 1598 503
700150581H1 SATMON007 g170772 BLASTN 640 1e-44 88 1599 503
700151970H1 SATMON007 g170772 BLASTN 645 1e-44 87 1600 503
700151780H1 SATMON007 g170772 BLASTN 628 1e-43 88 1601 503
700575547H1 SATMON030 g170772 BLASTN 614 1e-42 90 1602 503
LIB143-026-Q1-E1-A5 LIB143 g170772 BLASTN 633 1e-42 77 1603 503
700347945H1 SATMON023 g170772 BLASTN 586 1e-40 84 1604 503
700074416H1 SATMON007 g170772 BLASTN 589 1e-40 86 1605 503
700352990H1 SATMON024 g170772 BLASTN 575 1e-39 92 1606 503
700156025H1 SATMON007 g170772 BLASTN 568 1e-38 91 1607 503
700432072H1 SATMONN01 g170772 BLASTN 374 1e-37 84 1608 503
700354249H1 SATMON024 g170772 BLASTN 528 1e-35 93 1609 503
700617977H1 SATMON033 g170772 BLASTN 535 1e-35 87 1610 503
700218312H1 SATMON016 g170772 BLASTN 236 1e-33 91 1611 503
700456080H1 SATMON029 g170772 BLASTN 513 1e-33 83 1612 503
700349395H1 SATMON023 g170772 BLASTN 500 1e-32 91 1613 503
700051637H1 SATMON003 g170772 BLASTN 249 1e-31 86 1614 503
700202153H1 SATMON003 g170772 BLASTN 474 1e-30 91 1615 503
700256951H1 SATMON017 g170772 BLASTN 454 1e-29 86 1616 503
700150542H1 SATMON007 g170772 BLASTN 343 1e-28 76 1617 503
700151629H1 SATMON007 g170772 BLASTN 445 1e-28 85 1618 503
700446654H1 SATMON027 g170772 BLASTN 437 1e-27 84 1619 503
700155814H1 SATMON007 g170772 BLASTN 428 1e-26 88 1620 503
700161019H1 SATMON012 g2588780 BLASTN 417 1e-25 92 1621 503
700377236H1 SATMON019 g170772 BLASTN 402 1e-24 84 1622 503
LIB3067-036-Q1-K1-A4 LIB3067 g1220121 BLASTN 224 1e-22 84 1623 503
700158933H1 SATMON012 g170772 BLASTN 368 1e-21 90 1624 503
700155365H1 SATMON007 g170772 BLASTN 347 1e-20 91 1625 503
700405366H1 SATMON029 g170772 BLASTN 311 1e-17 88 1626 503
700159812H1 SATMON012 g407412 BLASTX 160 1e-15 96 1627 503
700209759H1 SATMON016 g170772 BLASTN 239 1e-15 90 1628 503
700154527H1 SATMON007 g170772 BLASTN 250 1e-12 92 1629 503
700449637H1 SATMON028 g170772 BLASTN 201 1e-10 78 1630 503
700096829H1 SATMON008 g170772 BLASTN 216 1e-9 89 2993 -700661285
700661285H1 SOYMON005 g1857024 BLASTX 95 1e-12 100 2994 -700750570
700750570H1 SOYMON014 g170772 BLASTN 414 1e-24 81 2995 -700752735
700752735H1 SOYMON014 g170772 BLASTN 446 1e-27 78 2996 -700755052
700755052H1 SOYMON014 g170772 BLASTN 547 1e-45 74 2997 -700756501
700756501H1 SOYMON014 g535583 BLASTN 717 1e-50 84 2998 -700831127
700831127H1 SOYMON019 g535583 BLASTN 862 1e-63 83 2999 -700851779
700851779H1 SOYMON023 g170772 BLASTN 505 1e-33 77 3000 -700888715
700888715H1 SOYMON024 g535583 BLASTN 442 1e-31 91
3001 -700889420 700889420H1 SOYMON024 g1220121 BLASTN 893 1e-65 84
3002 -700895218 700895218H1 SOYMON024 g407411 BLASTN 816 1e-59 83
3003 -700941379 700941379H1 SOYMON024 g170772 BLASTN 424 1e-33 72
3004 -700986855 700986855H1 SOYMON009 g170772 BLASTN 701 1e-49 74
3005 -701070484 701070484H1 SOYMON034 g407411 BLASTN 362 1e-43 73
3006 -701136279 701136279H1 SOYMON038 g170772 BLASTN 651 1e-56 80
3007 -GM16478 LIB3054-007-Q1-N1-G3 LIB3054 g2244750 BLASTX 70 1e-27
61 3008 -GM23819 LIB3040-019-Q1-E1-C5 LIB3040 g535583 BLASTN 258
1e-10 88 3009 -GM29758 LIB3050-016-Q1-E1-B11 LIB3050 g535583 BLASTN
391 1e-42 70 3010 16 LIB3030-003-Q1-B1-B11 LIB3030 g3088578 BLASTN
1577 1e-122 87 3011 16 LIB3050-023-Q1-K1-H9 LIB3050 g535583 BLASTN
1498 1e-116 86 3012 16 LIB3030-003-Q1-B1-F7 LIB3030 g535583 BLASTN
1469 1e-113 86 3013 16 LIB3055-005-Q1-N1-C11 LIB3055 g170772 BLASTN
1205 1e-109 84 3014 16 LIB3065-011-Q1-N1-A3 LIB3065 g170772 BLASTN
622 1e-107 88 3015 16 700652256H1 SOYMON003 g170772 BLASTN 591
1e-90 88 3016 16 LIB3065-011-Q1-N1-A4 LIB3065 g170772 BLASTN 1182
1e-89 78 3017 16 700653827H1 SOYMON003 g170772 BLASTN 660 1e-87 81
3018 16 701099940H1 SOYMON028 g1220121 BLASTN 1154 1e-87 88 3019 16
701003671H1 SOYMON019 g170772 BLASTN 1131 1e-85 92 3020 16
700752105H1 SOYMON014 g170772 BLASTN 1116 1e-84 91 3021 16
700653057H1 SOYMON003 g170772 BLASTN 990 1e-83 88 3022 16
700945531H1 SOYMON024 g535583 BLASTN 1109 1e-83 89 3023 16
700980013H1 SOYMON009 g535583 BLASTN 1109 1e-83 88 3024 16
701127812H1 SOYMON037 g170772 BLASTN 1110 1e-83 91 3025 16
LIB3056-014-Q1-N1-F8 LIB3056 g170772 BLASTN 798 1e-81 82 3026 16
700653862H1 SOYMON003 g1220121 BLASTN 960 1e-80 88 3027 16
700994148H1 SOYMON011 g535583 BLASTN 1069 1e-80 86 3028 16
700984184H1 SOYMON009 g170772 BLASTN 756 1e-79 86 3029 16
701123715H1 SOYMON037 g1220121 BLASTN 1059 1e-79 87 3030 16
700839038H1 SOYMON020 g170772 BLASTN 1060 1e-79 93 3031 16
700978445H1 SOYMON009 g170772 BLASTN 1062 1e-79 89 3032 16
701123035H1 SOYMON037 g170772 BLASTN 829 1e-78 90 3033 16
701041545H1 SOYMON029 g170772 BLASTN 1030 1e-77 86 3034 16
700898192H1 SOYMON027 g1220121 BLASTN 1031 1e-77 89 3035 16
700985750H1 SOYMON009 g169662 BLASTN 1033 1e-77 85 3036 16
700730995H1 SOYMON009 g170772 BLASTN 1040 1e-77 91 3037 16
700555909H1 SOYMON001 g535583 BLASTN 593 1e-76 84 3038 16
700746538H1 SOYMON013 g535583 BLASTN 834 1e-76 88 3039 16
700941380H1 SOYMON024 g170772 BLASTN 991 1e-76 91 3040 16
701118851H1 SOYMON037 g170772 BLASTN 1019 1e-76 89 3041 16
701065379H1 SOYMON034 g535583 BLASTN 1022 1e-76 86 3042 16
701209645H1 SOYMON035 g170772 BLASTN 795 1e-75 86 3043 16
701055017H1 SOYMON032 g1220121 BLASTN 877 1e-75 86 3044 16
701015213H1 SOYMON019 g535583 BLASTN 1008 1e-75 87 3045 16
700982770H1 SOYMON009 g1220121 BLASTN 1009 1e-75 85 3046 16
701212420H1 SOYMON035 g535583 BLASTN 1012 1e-75 86 3047 16
700977916H1 SOYMON009 g170772 BLASTN 700 1e-74 89 3048 16
700974401H1 SOYMON005 g1220121 BLASTN 857 1e-74 89 3049 16
700645749H1 SOYMON010 g170772 BLASTN 996 1e-74 82 3050 16
700646620H1 SOYMON014 g170772 BLASTN 996 1e-74 89 3051 16
701126103H1 SOYMON037 g170772 BLASTN 1000 1e-74 88 3052 16
700978001H1 SOYMON009 g170772 BLASTN 563 1e-73 89 3053 16
700561987H1 SOYMON002 g1220121 BLASTN 782 1e-73 87 3054 16
701101526H1 SOYMON028 g1220121 BLASTN 810 1e-73 88 3055 16
700562680H1 SOYMON002 g170772 BLASTN 985 1e-73 87 3056 16
700560521H1 SOYMON001 g170772 BLASTN 988 1e-73 80 3057 16
701055544H1 SOYMON032 g170772 BLASTN 992 1e-73 88 3058 16
701061418H1 SOYMON033 g170772 BLASTN 993 1e-73 87 3059 16
701049514H1 SOYMON032 g1220121 BLASTN 884 1e-72 88 3060 16
700646215H1 SOYMON012 g170772 BLASTN 971 1e-72 89 3061 16
701014875H1 SOYMON019 g535583 BLASTN 973 1e-72 87 3062 16
700874718H1 SOYMON018 g169662 BLASTN 974 1e-72 86 3063 16
700897796H1 SOYMON027 g1220121 BLASTN 977 1e-72 87 3064 16
700548019H1 SOYMON001 g170772 BLASTN 547 1e-71 87 3065 16
700904875H1 SOYMON022 g535583 BLASTN 963 1e-71 88 3066 16
701106315H1 SOYMON036 g170772 BLASTN 557 1e-70 90 3067 16
700745023H1 SOYMON013 g407411 BLASTN 949 1e-70 85 3068 16
701120348H1 SOYMON037 g170772 BLASTN 956 1e-70 86 3069 16
701124508H1 SOYMON037 g170772 BLASTN 578 1e-69 84 3070 16
700956183H1 SOYMON022 g1220121 BLASTN 625 1e-69 87 3071 16
700894619H1 SOYMON024 g535583 BLASTN 828 1e-69 85 3072 16
700892392H1 SOYMON024 g535583 BLASTN 936 1e-69 85 3073 16
700730676H1 SOYMON009 g535583 BLASTN 939 1e-69 86 3074 16
700728913H1 SOYMON009 g407411 BLASTN 940 1e-69 87 3075 16
700845737H1 SOYMON021 g1220121 BLASTN 528 1e-68 88 3076 16
700990961H1 SOYMON011 g170772 BLASTN 582 1e-68 85 3077 16
700983443H1 SOYMON009 g170772 BLASTN 922 1e-68 86 3078 16
701120754H1 SOYMON037 g170772 BLASTN 923 1e-68 88 3079 16
700900486H1 SOYMON027 g1220121 BLASTN 924 1e-68 83 3080 16
701133574H2 SOYMON038 g170772 BLASTN 929 1e-68 88 3081 16
700900924H1 SOYMON027 g170772 BLASTN 931 1e-68 88 3082 16
701056706H1 SOYMON032 g170772 BLASTN 486 1e-67 89 3083 16
701110051H1 SOYMON036 g170772 BLASTN 910 1e-67 88 3084 16
701136325H1 SOYMON038 g170772 BLASTN 915 1e-67 87 3085 16
700750809H1 SOYMON014 g170772 BLASTN 900 1e-66 88 3086 16
700686607H1 SOYMON008 g170772 BLASTN 901 1e-66 88 3087 16
700848261H1 SOYMON021 g535583 BLASTN 905 1e-66 87 3088 16
700686634H1 SOYMON008 g170772 BLASTN 905 1e-66 88 3089 16
700891285H1 SOYMON024 g170772 BLASTN 906 1e-66 88 3090 16
700560291H1 SOYMON001 g170772 BLASTN 906 1e-66 88 3091 16
700752975H1 SOYMON014 g170772 BLASTN 907 1e-66 89 3092 16
701006013H2 SOYMON019 g170772 BLASTN 908 1e-66 87 3093 16
700974038H1 SOYMON005 g1220121 BLASTN 631 1e-65 88 3094 16
701047024H1 SOYMON032 g170772 BLASTN 707 1e-65 89 3095 16
700900409H1 SOYMON027 g170772 BLASTN 737 1e-65 83 3096 16
701137320H1 SOYMON038 g170772 BLASTN 769 1e-65 88 3097 16
700978805H1 SOYMON009 g170772 BLASTN 886 1e-65 84 3098 16
700726195H1 SOYMON009 g1220121 BLASTN 889 1e-65 87 3099 16
700661112H1 SOYMON005 g170772 BLASTN 661 1e-64 85 3100 16
700989712H1 SOYMON011 g170772 BLASTN 788 1e-64 88 3101 16
700752287H1 SOYMON014 g170772 BLASTN 875 1e-64 85 3102 16
700964226H1 SOYMON022 g170772 BLASTN 877 1e-64 88 3103 16
700847346H1 SOYMON021 g170772 BLASTN 880 1e-64 83 3104 16
700756428H1 SOYMON014 g170772 BLASTN 883 1e-64 88 3105 16
701049928H1 SOYMON032 g170772 BLASTN 885 1e-64 82 3106 16
700848652H1 SOYMON021 g170772 BLASTN 477 1e-63 89 3107 16
700898929H1 SOYMON027 g1220121 BLASTN 514 1e-63 87 3108 16
700903523H1 SOYMON022 g170772 BLASTN 527 1e-63 82 3109 16
700983745H1 SOYMON009 g1220121 BLASTN 863 1e-63 88 3110 16
700890587H1 SOYMON024 g170772 BLASTN 865 1e-63 86 3111 16
700969917H1 SOYMON005 g407411 BLASTN 868 1e-63 83 3112 16
700808487H1 SOYMON024 g170772 BLASTN 870 1e-63 88 3113 16
700749968H1 SOYMON013 g170772 BLASTN 871 1e-63 87 3114 16
700751254H1 SOYMON014 g170772 BLASTN 575 1e-62 87 3115 16
701014277H1 SOYMON019 g170772 BLASTN 739 1e-62 86 3116 16
700853635H1 SOYMON023 g170772 BLASTN 854 1e-62 87 3117 16
700752357H1 SOYMON014 g170772 BLASTN 856 1e-62 88 3118 16
700754523H1 SOYMON014 g170772 BLASTN 856 1e-62 92 3119 16
700982153H1 SOYMON009 g170772 BLASTN 400 1e-61 82 3120 16
700958283H1 SOYMON022 g170772 BLASTN 839 1e-61 87 3121 16
700980911H1 SOYMON009 g170772 BLASTN 842 1e-61 82 3122 16
700788112H1 SOYMON011 g170772 BLASTN 844 1e-61 83 3123 16
701005927H1 SOYMON019 g170772 BLASTN 849 1e-61 88 3124 16
700756443H1 SOYMON014 g170772 BLASTN 849 1e-61 88 3125 16
700658914H1 SOYMON004 g1220121 BLASTN 468 1e-60 85 3126 16
701135266H1 SOYMON038 g170772 BLASTN 827 1e-60 87 3127 16
700982179H1 SOYMON009 g170772 BLASTN 827 1e-60 82 3128 16
700754593H1 SOYMON014 g170772 BLASTN 832 1e-60 81 3129 16
700831723H1 SOYMON019 g170772 BLASTN 814 1e-59 91 3130 16
700986775H1 SOYMON009 g170772 BLASTN 815 1e-59 92 3131 16
700755219H1 SOYMON014 g170772 BLASTN 820 1e-59 84 3132 16
701015494H1 SOYMON019 g170772 BLASTN 822 1e-59 88 3133 16
701008473H1 SOYMON019 g170772 BLASTN 823 1e-59 87 3134 16
700754981H1 SOYMON014 g170772 BLASTN 824 1e-59 88 3135 16
700870790H1 SOYMON018 g170772 BLASTN 825 1e-59 81 3136 16
700833069H1 SOYMON019 g170772 BLASTN 806 1e-58 86 3137 16
700680127H2 SOYMON008 g535583 BLASTN 807 1e-58 86 3138 16
701015374H1 SOYMON019 g170772 BLASTN 807 1e-58 87 3139 16
700872895H1 SOYMON018 g535583 BLASTN 808 1e-58 88 3140 16
701137912H1 SOYMON038 g170772 BLASTN 374 1e-57 83 3141 16
700984063H1 SOYMON009 g1220121 BLASTN 575 1e-57 79 3142 16
700991988H1 SOYMON011 g170772 BLASTN 791 1e-57 82 3143 16
700873915H1 SOYMON018 g170772 BLASTN 793 1e-57 88 3144 16
700978721H1 SOYMON009 g170772 BLASTN 797 1e-57 78 3145 16
701213679H1 SOYMON035 g170772 BLASTN 798 1e-57 88 3146 16
701102954H1 SOYMON028 g170772 BLASTN 799 1e-57 79 3147 16
700888289H1 SOYMON024 g170772 BLASTN 712 1e-56 91 3148 16
700962086H1 SOYMON022 g170772 BLASTN 783 1e-56 88 3149 16
701052227H1 SOYMON032 g535583 BLASTN 788 1e-56 86 3150 16
700755177H1 SOYMON014 g170772 BLASTN 768 1e-55 88 3151 16
701123136H1 SOYMON037 g170772 BLASTN 771 1e-55 82 3152 16
700979067H1 SOYMON009 g170772 BLASTN 776 1e-55 90 3153 16
700755513H1 SOYMON014 g170772 BLASTN 759 1e-54 92 3154 16
700756639H1 SOYMON014 g170772 BLASTN 761 1e-54 81 3155 16
700554077H1 SOYMON001 g170772 BLASTN 371 1e-53 86 3156 16
700653194H1 SOYMON003 g170772 BLASTN 395 1e-53 86 3157 16
701110348H1 SOYMON036 g170772 BLASTN 743 1e-53 81 3158 16
700753973H1 SOYMON014 g170772 BLASTN 743 1e-53 87 3159 16
701011009H1 SOYMON019 g535583 BLASTN 733 1e-52 81 3160 16
700739086H1 SOYMON012 g170772 BLASTN 456 1e-51 87 3161 16
700740140H1 SOYMON012 g170772 BLASTN 723 1e-51 89 3162 16
700565040H1 SOYMON002 g170772 BLASTN 729 1e-51 74 3163 16
701148186H1 SOYMON031 g535583 BLASTN 665 1e-50 85 3164 16
701142734H1 SOYMON038 g535583 BLASTN 670 1e-47 86 3165 16
700754441H1 SOYMON014 g170772 BLASTN 385 1e-46 93 3166 16
701102588H1 SOYMON028 g1220121 BLASTN 662 1e-46 89 3167 16
700900656H1 SOYMON027 g535583 BLASTN 635 1e-44 87 3168 16
700974207H1 SOYMON005 g535583 BLASTN 641 1e-44 85 3169 16
700982081H1 SOYMON009 g170772 BLASTN 494 1e-43 78 3170 16
701130026H1 SOYMON037 g1220121 BLASTN 482 1e-42 86 3171 16
701009720H1 SOYMON019 g170772 BLASTN 621 1e-42 87 3172 16
700962602H1 SOYMON022 g170772 BLASTN 357 1e-40 91 3173 16
700724934H1 SOYMON009 g535583 BLASTN 561 1e-40 81 3174 16
700729305H1 SOYMON009 g535583 BLASTN 544 1e-39 82 3175 16
701210054H1 SOYMON035 g535583 BLASTN 569 1e-38 85 3176 16
700790192H1 SOYMON011 g535583 BLASTN 293 1e-37 83 3177 16
700984076H1 SOYMON009 g170772 BLASTN 198 1e-35 88 3178 16
700726562H1 SOYMON009 g170772 BLASTN 307 1e-34 78 3179 16
701211376H1 SOYMON035 g170772 BLASTN 524 1e-34 86 3180 16
700753085H1 SOYMON014 g2588780 BLASTN 358 1e-33 76 3181 16
700727993H1 SOYMON009 g535583 BLASTN 465 1e-32 84 3182 16
700561072H1 SOYMON001 g170772 BLASTN 473 1e-30 83 3183 16
701211464H1 SOYMON035 g170772 BLASTN 464 1e-28 81 3184 16
701098045H1 SOYMON028 g169660 BLASTN 386 1e-21 78 3185 16
700945233H1 SOYMON024 g407412 BLASTX 150 1e-18 87 3186 16
700752655H1 SOYMON014 g758247 BLASTX 172 1e-16 94 3187 16
700735356H1 SOYMON010 g758247 BLASTX 152 1e-14 100 3188 16
700683995H1 SOYMON008 g758247 BLASTX 106 1e-13 90 3189 16
700658760H1 SOYMON004 g1857024 BLASTX 123 1e-13 63 3190 16
700762885H1 SOYMON015 g1857024 BLASTX 134 1e-13 89 3191 16
700755740H1 SOYMON014 g170773 BLASTX 149 1e-13 100 3192 16
700854969H1 SOYMON023 g170772 BLASTN 178 1e-12 80 3193 16
701143036H1 SOYMON038 g169661 BLASTX 113 1e-8 83 3194 18409
700786561H1 SOYMON011 g535583 BLASTN 992 1e-73 85 3195 18409
701008057H1 SOYMON019 g535583 BLASTN 831 1e-60 87 3196 18409
701037442H1 SOYMON029 g535583 BLASTN 669 1e-46 86 3197 18409
700942865H1 SOYMON024 g535583 BLASTN 464 1e-32 86 3198 7322
700651524H1 SOYMON003 g170772 BLASTN 466 1e-65 80 3199 7322
700565758H1 SOYMON002 g170772 BLASTN 450 1e-63 81 CYSTATHIONINE
.beta.-SYNTHASE (EC 4.2.1.22) 1631 -700025795 700025795H1 SATMON003
g1323263 BLASTX 186 1e-25 68 1632 20651 700344783H1 SATMON021
g1813975 BLASTX 41 1e-9 53 CYSTATHIONINE .gamma.-LYASE (EC 4.4.1.1)
1633 -700260027 700260027H1 SATMON017 g169475 BLASTX 112 1e-10 75
1634 1228 700027629H1 SATMON003 g169475 BLASTX 189 1e-19 87 3203
-700750583 700750583H1 SOYMON014 g169475 BLASTX 149 1e-13 78 3204
12502 LIB3051-069-Q1-K1-E6 LIB3051 g2641242 BLASTX 86 1e-30 38
O-ACETYLHOMOSERINE (THIOL)-LYASE (EC 4.2.99.10) 3200 12502
701135185H1 SOYMON038 g1628606 BLASTX 100 1e-10 48 3201 12502
701042913H1 SOYMON029 g2605905 BLASTX 110 1e-9 42 3202 12502
701059330H1 SOYMON033 g2605905 BLASTX 110 1e-9 42
Table Headings
Cluster ID
[0499] A cluster ID is arbitrarily assigned to all of those clones
which belong to the same cluster at a given stringency and a
particular clone will belong to only one cluster at a given
stringency. If a cluster contains only a single clone (a
"singleton"), then the cluster ID number will be negative, with an
absolute value equal to the clone ID number of its single member.
The cluster ID entries in the table refer to the cluster with which
the particular clone in each row is associated.
Clone ID
[0500] The clone ID number refers to the particular clone in the
PhytoSeq database. Each clone ID entry in the table refers to the
clone whose sequence is used for (1) the sequence comparison whose
scores are presented and/or (2) assignment to the particular
cluster which is presented. Note that a clone may be included in
this table even if its sequence comparison scores fail to meet the
minimum standards for similarity. In such a case, the clone is
included due solely to its association with a particular cluster
for which sequences of one or more other member clones possess the
required level of similarity.
Library
[0501] The library ID refers to the particular cDNA library from
which a given clone is obtained. Each cDNA library is associated
with the particular tissue(s), line(s) and developmental stage(s)
from which it is isolated.
NCBI gi
[0502] Each sequence in the GenBank public database is arbitrarily
assigned a unique NCBI gi (National Center for Biotechnology
Information GenBank Identifier) number. In this table, the NCBI gi
number which is associated (in the same row) with a given clone
refers to the particular GenBank sequence which is used in the
sequence comparison. This entry is omitted when a clone is included
solely due to its association with a particular cluster.
Method
[0503] The entry in the "Method" column of the table refers to the
type of BLAST search that is used for the sequence comparison.
"CLUSTER" is entered when the sequence comparison scores for a
given clone fail to meet the minimum values required for
significant similarity. In such cases, the clone is listed in the
table solely as a result of its association with a given cluster
for which sequences of one or more other member clones possess the
required level of similarity.
Score
[0504] Each entry in the "Score" column of the table refers to the
BLAST score that is generated by sequence comparison of the
designated clone with the designated GenBank sequence using the
designated BLAST method. This entry is omitted when a clone is
included solely due to its association with a particular cluster.
If the program used to determine the hit is HMMSW then the score
refers to HMMSW score.
P-Value
[0505] The entries in the P-Value column refer to the probability
that such matches occur by chance.
% Ident
[0506] The entries in the "% Ident" column of the table refer to
the percentage of identically matched nucleotides (or residues)
that exist along the length of that portion of the sequences which
is aligned by the BLAST comparison to generate the statistical
scores presented. This entry is omitted when a clone is included
solely due to its association with a particular cluster.
Sequence CWU 0 SQTB SEQUENCE LISTING The patent application
contains a lengthy "Sequence Listing" section. A copy of the
"Sequence Listing" is available in electronic form from the USPTO
web site
(http://seqdata.uspto.gov/?pageRequest=docDetail&DocID=US20100071097A1).
An electronic copy of the "Sequence Listing" will also be available
from the USPTO upon request and payment of the fee set forth in 37
CFR 1.19(b)(3).
0 SQTB SEQUENCE LISTING The patent application contains a lengthy
"Sequence Listing" section. A copy of the "Sequence Listing" is
available in electronic form from the USPTO web site
(http://seqdata.uspto.gov/?pageRequest=docDetail&DocID=US20100071097A1).
An electronic copy of the "Sequence Listing" will also be available
from the USPTO upon request and payment of the fee set forth in 37
CFR 1.19(b)(3).
* * * * *
References