U.S. patent application number 10/651991 was filed with the patent office on 2005-06-09 for differentially-expressed conifer cdnas, and their use in improving somatic embryogenesis.
This patent application is currently assigned to INSTITUTE OF PAPER SCIENCE AND TECHNOLOGY, INC., INSTITUTE OF PAPER SCIENCE AND TECHNOLOGY, INC.. Invention is credited to Cairney, John, Xu, Nanfei.
Application Number | 20050125161 10/651991 |
Document ID | / |
Family ID | 34637088 |
Filed Date | 2005-06-09 |
United States Patent
Application |
20050125161 |
Kind Code |
A1 |
Cairney, John ; et
al. |
June 9, 2005 |
Differentially-expressed conifer cDNAs, and their use in improving
somatic embryogenesis
Abstract
The invention relates to a method for staging embryos of plants.
In particular, this invention relates to a method for creating a
relational database by determining transcript levels of sets of
genes expressed at predetermined stages in embryo development. This
approach creates a method by which embryos of unknown stage
development can be determined by comparisons between expression
levels of those embryos to the expression levels found in the
database. This approach further allows rapid identification of
transcripts in an embryo to be staged by the utilization of probes
corresponding to cDNAs comprising the database. Additionally, this
invention relates to a method for selecting advantageous plant
clones for future propagation. Specifically, this method relates to
an approach to link the biochemical condition of an embryo to
current culture conditions and thus provides a method for enhancing
conditions to produce embryos with a desired biochemical state.
Inventors: |
Cairney, John; (Decatur,
GA) ; Xu, Nanfei; (Wildwood, MO) |
Correspondence
Address: |
FINNEGAN, HENDERSON, FARABOW, GARRETT & DUNNER
LLP
1300 I STREET, NW
WASHINGTON
DC
20005
US
|
Assignee: |
INSTITUTE OF PAPER SCIENCE AND
TECHNOLOGY, INC.
|
Family ID: |
34637088 |
Appl. No.: |
10/651991 |
Filed: |
September 2, 2003 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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10651991 |
Sep 2, 2003 |
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09973994 |
Oct 11, 2001 |
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60239250 |
Oct 11, 2000 |
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60260882 |
Jan 12, 2001 |
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Current U.S.
Class: |
702/20 |
Current CPC
Class: |
G16B 25/00 20190201;
G16B 25/10 20190201; G16B 50/00 20190201; C12Q 1/6895 20130101 |
Class at
Publication: |
702/020 |
International
Class: |
G06F 019/00; G01N
033/48; G01N 033/50 |
Claims
1. A relational database comprising the data of Table I.
2. A method of staging embryos comprising: a) providing at least
one embryo; b) detecting the expression in the embryo of at least
one RNA transcript of Table I; and c) correlating the expression of
said transcript to one or more embryonic stages.
3. The method of claim 2 wherein at least two RNA transcripts are
detected or determined and correlated to one or more embryonic
stages.
4. The method of claim 2 wherein expression of the at least one RNA
transcript is analyzed by hybridization with at least one probe of
Table I.
5. The method of claim 2 wherein expression of the at least one RNA
transcript is analyzed by hybridization with a variant of at least
one probe of Table I.
6. The method of claim 5 wherein said variant hybridizes to at
least one probe of Table I under conditions of high stringency.
7. The method of claim 5 wherein said variant hybridizes to at
least one probe of Table I under conditions of moderate
stringency.
8. The method of claim 2 wherein expression of at least one RNA
transcript is detected or determined by at least one member of the
group consisting of PCR, Northern Analysis, and in situ
hybridization.
9. The method of claim 2 wherein expression of said at least two
RNA transcripts are detected by a DNA array.
10. A database comprising a multiplicity of nucleotide sequences
shown in any one of Table I, including variants thereof, wherein
said variants hybridize under conditions of high stringency to
either strand of a denatured, double-stranded DNA comprising any of
SEQ ID NOS: 1-327.
11. The database of claim 10 wherein said variants hybridize under
conditions of moderate stringency.
12. A DNA array comprising a multiplicity of nucleotide sequences
shown in Table I, including variants thereof, wherein said variants
hybridize under conditions of high stringency to either strand of a
denatured, double-stranded DNA comprising any of SEQ ID NOS:
1-327.
13. The DNA array of claim 12 wherein said variants hybridize under
conditions of moderate stringency.
14. A method for staging plant embryos comprising: a) selecting
total RNA from a multiplicity of embryos of known developmental
age; b) correlating the embryonic expression pattern to the
developmental age to develop a relational database; c) determining
levels of expression from embryos of unknown developmental age by
hybridization to a DNA array comprising a multiplicity of the
nucleotide sequences shown in any one of SEQ ID NOS: 1-327; d)
correlating the expression pattern from step 3 to the relational
database to determine developmental stage for the unknown
embryo.
15. The method of claim 14 wherein the embryos of step 1) are
zygotic embryos.
16. The method of claim 14 further comprising the step of altering
the embryonic growth conditions to approximate the expression
pattern of zygotic embryos.
17. An isolated nucleic acid variant of the nucleotide sequence
shown in any one of SEQ ID NOS: 1-334, wherein said variant
hybridizes under conditions of moderate stringency to either strand
of a denatured, double-stranded DNA comprising any of SEQ ID NOS:
1-334.
18. An isolated polypeptide encoded by a nucleic acid molecule of
claim 17.
19. An isolated nucleic acid encoding the polypeptide of claim
18.
20. Antibodies that specifically bind to the peptide of claim
18.
21. The antibodies of claim 20, wherein said antibodies are
monoclonal.
22. A recombinant vector that directs the expression of a nucleic
acid of claim 17.
23. A host cell transformed with the vector of claim 22.
24. The host cell of claim 23, wherein the host is a somatic pine
embryo.
25. A method for staging plant embryos comprising: a) selecting
total RNA from at least one embryo of known developmental age; b)
determining the level of expression of a multiplicity of genes
which hybridize to one or more of SEQ ID NOS: 1-327; c) correlating
the known developmental ages of the embryos from step 1) with the
profile of expression measured in step 2); d) applying the
correlation of step 3) to a sample of embryo RNA from embryos to be
staged; and e) determining the embryo stage.
26. The method of claim 25, wherein the measurement of gene
expression is by RT-PCR.
27. The method of claim 25, wherein the measurement of gene
expression is by nucleic acid hybridization.
28. The method of claim 25, wherein the measurement of gene
expression is by determining the level of protein expression.
29. The method of claim 28, wherein protein expression is measured
by antibody binding.
30. A method for selecting advantageous plant clones comprising: a)
selecting one or more samples of embryonic RNA from multiple clones
of plants; b) determining that at least one sampled clone has an
advantageous characteristic; c) comparing the embryonic levels of
expression of genes which hybridize to one or more of SEQ ID NOS:
1-327 in samples from the advantageous clone with expression levels
in at least one clone that does not show the advantageous
characteristic; and d) selecting additional clones which show an
embryonic gene expression pattern more similar to that of the
advantageous clone than to the pattern of at least one clone that
does not show the advantageous characteristic.
31. Method of claim 30 where the clones to be sampled or compared
are from about the same developmental age.
32. Method of claim 31 where the development age is visually
detected.
33. The method of claim 30, wherein the measurement of gene
expression is by RT-PCR.
34. The method of claim 30, wherein the measurement of gene
expression is by nucleic acid hybridization.
35. The method of claim 30, wherein the measurement of gene
expression is by determining the level of protein expression.
36. The method of claim 35, wherein protein expression is measured
by antibody binding.
37. A method of determining embryo fitness comprising: a) creating
a relational database with RNA expression values for genes listed
in Table I for embryos of known developmental stages; b) isolating
total RNA from embryos of unknown stage development; c) measuring
expression levels of genes identified in Table I from the solated
total RNA; and d) correlating the database of step 1) with the
pattern of expression determined in steps 2) and 3) to assess
proper embryo development.
38. The method of claim 37, wherein the measurement of gene
expression is by RT-PCR.
39. The method of claim 37, wherein the measurement of gene
expression is by nucleic acid hybridization.
40. The method of claim 37, wherein the measurement of gene
expression is by determining the level of protein expression.
41. The method of claim 40, wherein protein expression is measured
by antibody binding.
42. A method for selecting advantageous growth conditions for
embryo development comprising: a) determining RNA expression
profiles for staged embryos under control culture conditions; b)
altering culture conditions; c) determining RNA expression profiles
for staged embryos under altered culture conditions; and d)
correlating culture change to developmental effect in embryo.
43. The method of claim 42, wherein conditions are selected which
produce RNA expression profiles most closely approximating
late-stage embryo profiles.
44. The method of claim 42, wherein the culture conditions are
altered by operatively linking one or more stage-specific embryo
promoter(s) to one or more sense or antisense nucleic acid
molecules.
45. The method of claim 42, wherein the culture conditions are
altered by operatively linking one more stage-specific embryo
promoter(s) selected from SEQ ID NOS: 328-334 to one or more sense
or antisense nucleic acid molecules.
46. The method of claim 42, wherein the change in expression
profiles is correlated by a relational database.
47. A recombinant nucleic acid molecule encoding a product during
embryo development comprising: a) a first nucleic acid sequence
which is the LP2-3 promoter; and b) a second nucleic acid sequence
encoding a product, wherein the first nucleic acid is operatively
linked to the second nucleic acid molecule whereby its expression
is directed by the promoter sequence.
48. The recombinant nucleic acid molecule of claim 47 wherein the
second nucleic acid sequence encodes for GFP, or a variant of
GFP.
49. The recombinant nucleic acid molecule of claim 48 wherein the
second nucleic acid sequence is linked to one or more additional
nucleic acid molecules.
50. The recombinant nucleic acid molecule of claim 49 wherein the
additional molecule encodes a protein product normally expressed,
by a developing embryo at a known stage.
51. The recombinant nucleic acid molecule of claim 47 wherein the
second nucleic acid sequence encodes an embryo-derived
molecule.
52. The recombinant nucleic acid molecule of claim 51
embryo-derived molecule is stage-specific.
53. A plant cell comprising the recombinant nucleic acid molecule
of claim 47.
54. A method for producing a protein product during embryo
development comprising: a) operatively linking one more
stage-specific embryo promoter(s) to one or more nucleic acid
molecules that encode a protein product, b) delivering construct to
developing embryos.
55. The method of claim 54 wherein the operatively linked nucleic
acid molecule is a reporter or indicator gene.
56. The method of claim 54 wherein the operatively linked nucleic
acid molecule is GFP, or a variant of GFP
57. The method of claim 54 wherein at least one stage-specific
promoter is selected from SEQ ID NOS: 328-334.
58. A method for staging embryos comprising: a) providing one or
more stage-specific embryo promoter(s) operatively linked to one or
more nucleic acid molecules that encode a protein product to
developing embryos, b) monitoring expression of the protein product
as the embryo matures through stage in which promoter
functions.
59. The method of claim 58 wherein the operatively linked nucleic
acid molecule is a reporter or indicator gene.
60. The method of claim 58 wherein the operatively linked nucleic
acid molecule is GFP, or a variant of GFP.
61. The method of claim 58 wherein at least one stage-specific
promoter is selected from SEQ ID NOS: 328-334.
Description
CROSS-REFERENCE TO RELATED APPLICATIONS
[0001] This patent application claims benefit of priority of
provisional application U.S. Ser. No. 60/239,250, filed Oct. 11,
2000, and claims benefit of priority of provisional application
U.S. Ser. No. 60/260,882, filed Jan. 12, 2001.
FIELD OF THE INVENTION
[0002] The present invention relates to a relational database of
cDNA molecules, including those corresponding to Loblolly Pine
Major Intrinsic Protein (MIP), which are differentially expressed
during plant embryogenesis. The present invention further relates
to the use of DNA arrays for evaluating gene expression in somatic
and zygotic embryos. The invention encompasses related nucleic
acids, proteins, antigens, and antibodies derived from these cDNAs
as well as the use of such molecules for the staging,
characterization, and manipulation of plant embryogenesis, in
particular conifer embryogenesis. The cDNAs and related nucleic
acids, proteins, antigens, and antibodies derived from these cDNAs
are useful in the design, selection, and cultivation of improved
crops, specifically including coniferous trees, which provide raw
materials for paper and wood products.
BACKGROUND OF THE INVENTION
[0003] The world demand for paper is expected to increase nearly
50% by the year 2010 (McNutt and Rennel, Pulp Paper Intern 39: 48
(1997)). The United States' forest products industry faces a great
challenge in order to keep pace with the growing demand for paper.
This challenge is made greater by the decreasing availability of a
forest land-base, resulting from environmental restrictions and
urban growth, from which to harvest timber resources. Additionally,
valuable wood resources are lost to the environmental stresses and
biotic diseases. Consequently, the push to secure a renewable and
sustainable source of raw material for paper and other wood related
products has become an important priority for the forest products
industry.
[0004] Current forestry related research and development is focused
on creating sustainable fiber farms or tree plantations. Farming
trees with elite germplasms will increase growth rates and yields
of wood per acre. However, creating improved tree stock requires
the ability to identify and generate genetically superior trees and
a way to propagate such superior trees without diluting their
genetic quotient.
[0005] A. Breeding and Selection
[0006] Addressing the need to propagate genetically superior trees
without genetic diminution demands full research attention.
Traditional methods of tree propagation relied on selected breeding
programs to achieve genetic gain, i.e., the development of a
strain, sub-strain, or line having any heritable and economically
valuable characteristic or combination of characteristics not found
in the parents. Based on the results of progeny tests, superior
maternal trees are selected and used in "seed orchards" for mass
production of genetically improved seed. The genetic gain in such
an open-pollinated sexual propagation strategy is, however, limited
by the breeder's inability to control the paternal parent.
Additional gains can also be achieved by control-pollination of the
maternal tree with pollen from individual trees whose progeny have
demonstrated superior growth characteristics. Nevertheless, even
under controlled conditions where both parents of each seed are the
same, sexual propagation results in a "family" of seeds, i.e.,
siblings, comprised of many different genetic combinations. As not
all genotype combinations are favorable, the genetic gain in any
particular progeny is frequently offset and obscured by the genetic
variation among sibling seeds and those seedlings retaining
undesirable or previously masked pre-cross traits.
[0007] In addition to inherent genetic limitations of a traditional
breeding programs, large-scale production of control pollinated
seeds is also expensive. Consequently, economic and biological
limitations of large-scale seed production has lead the industry to
turn towards methods of asexual reproduction, such as grafting,
vegetative propagation and micropropagation, as more viable
alternatives.
[0008] B. Asexual (Clonal) Propagation
[0009] Asexual propagation permits the application of very high
selection intensity, resulting in the propagation of only those
progeny showing a high genetic gain potential. These highly
desirable progeny can have unique genetic combinations that result
in superior growth and performance characteristics. Thus, with
asexual propagation it is possible to genetically select
individuals while avoiding a concomitant reduction of genetic gain
due to intra-familial variation.
[0010] Asexual propagation of trees can be accomplished currently
by grafting, vegetative propagation, and micropropagation.
Grafting, widely used to propagate select individuals in limited
quantities for seed orchard establishment, is not applicable to
large-scale production for reforestation. Vegetative propagation,
achieved by the rooting of cuttings, and micropropagation by
somatic embryogenesis, currently hold the most potential for
reforestation of conifers. Although vegetative propagation by
rooted cuttings can be achieved in many coniferous species,
large-scale production via this method is extremely costly due to
difficulties in automating and mechanizing the process, not to
mention the need for tremendous quantities of stock tissue. This
propagation method is still further limited by the fact that the
rooting potential of stock plants decrease with time, making it
difficult to serially propagate from select genotypes over extended
periods of time.
[0011] Micropropagation is the most promising method of asexual
propagation for mass plantings. This process involves the
production of somatic embryos in vitro from minute pieces of plant
tissue or individual cells. The embryos are referred to as somatic
because they are derived from the somatic (vegetative) tissue,
rather than from the sexual process. Both vegetative propagation
and micropropagation have the potential to capture all genetic gain
of highly desirable genotypes. However, unlike conventional
vegetative propagation methods, somatic embryogenesis is amenable
to automation and mechanization, making it highly desirable for
large-scale production of planting stock for reforestation.
Moreover, somatic embryogenesis is particularly amenable to high
intensity selection of a large number of clones. These advantages
are compounded by the ability to safely preserve somatic
embryogenic cultures in liquid nitrogen for long-term storage.
Consequently, long-term cryogenic preservation offers immense
advantages over other vegetative propagation systems that attempt
to maintain the juvenility of stock plants. Techniques for somatic
embryogenesis in a wide variety of plant species are well known in
the art; exemplary methods for performing somatic embryogenesis in
conifers are taught in U.S. Pat. Nos.: 5,036,007; 5,236,841;
5,294,549; 5,413,930; 5,491,090; 5,506,136; 5,563,061; 5,677,185;
5,731,203; 5,731,204; and 5, 856,191, herein incorporated by
reference in their entirety.
[0012] Thus, somatic embryogenesis has great potential for clonal
production of conifer embryos to meet the increased demands of the
pulp and paper industry. Assessment of embryo quality, however,
needs improvement. The process of creating better tree stock begins
with understanding the process of tree development from
embryogenesis through full maturation.
[0013] In general, plant tissue culture is the broad science of
growing plant tissues on or in a nutrient medium containing
minerals, sugars, vitamins and plant hormones. By adjusting the
composition of the media, cultured tissues can be induced to grow
or differentiate into specific cell types or organs. "Somatic
embryogenesis" is a type of plant tissue culture where a piece of a
donor plant is excised, cultured and induced to form multiple
embryos. An embryo is a discrete mass of cells with a well-defined
structure that is capable of growing into a whole plant.
[0014] The methods generally in use for somatic embryogenesis today
involve several steps. Prior to the tissue culture process, a
suitable "explant" is harvested. A typical explant in conifer
somatic embryogenesis is the "megagametophyte", a haploid nutritive
tissue of the conifer seed, which is extracted from the ovule of a
pollinated female cone. This ovule contains single or multiple
zygotic seed embryos. In the seeds of many coniferous species, one
or more genetically unique embryos naturally undergo a process
called cleavage polyembryony, where a zygotic embryo grows and
divides to form a small clones of embryos.
[0015] The first step in somatic embryogenesis is the initiation
step. The explant is placed on a suitable media. When the explant
is an ovule, a process called extrusion occurs. Extrusion involves
the emergence or expulsion of a zygotic embryo or multiple embryos
and embryogenic tissue out of the megagametophyte. If culture
conditions are suitable, initiation proceeds and the extruded
embryo or embryos undergo the process of cleavage polyembryony.
This results in the formation of early stage somatic embryos in a
glossy, mucilaginous mass.
[0016] After embryogenic cultures are initiated, the somatic
embryos are transferred to a second medium with an appropriate
composition of plant hormones and other factors to induce the
somatic embryos to multiply. In the multiplication stage, cultures
can double up to 2-6 times in one week. Once large numbers of
embryos are obtained in the multiplication stage, the embryos are
moved to a development and maturation medium. Here, the correct
balance of plant hormones and other factors will induce the
early-stage embryos to mature into late stage embryos. Following
the maturation and development stage, embryos are germinated to
form small seedlings. These seedlings are then acclimated for
survival outside of the culture vessel. After acclimation, the
seedlings are ready for planting.
[0017] The relative ability to propagate plants by somatic
embryogenesis can vary greatly between species. Among conifers, for
example, spruce (Picea) species and Douglas fir are easily
propagated, while Pinus species are much more difficult. Many Pinus
species, including Loblolly pine (Pinus taeda), do not readily
initiate embryonic cultures. Typical initiation frequencies between
1% and 12% are reported for various Pinus species (Becwar et al.,
For. Sci. p1-18 (1988), Jain et al., Plant Sci. 65:233-241 (1989),
Becwar et al., Can. J. For. Res. 20:810 (1990), Li and Huang, J.
Tissue Cult. Assoc. 32:129 (1996)). Laine and David, (Plant Sci.
69:215 (1990)), however, were able to obtain high frequencies of
initiation (up to 59%) in Pinus caibaea, suggesting that not all
Pinus species are recalcitrant. Also, one earlier report described
initiation frequencies of 54% in White pine (Pinus strobus). Finer
et al., Plant Cell Rep. 8:203 (1989). However, other workers were
not able to duplicate this success. Michler et al., Plant Sci.
77:111 (1991). The results in the literature demonstrate the
recalcitrance of Pinus species, especially Loblolly pine, in
regeneration by somatic embryogenesis.
[0018] Nevertheless, once this process is understood from the
standpoint of developmental genetics, breeders will then have the
appropriate tools to monitor, intervene, and improve both the
regeneration frequency and the overall quality of tree stock
through genetic engineering. For example, both environmental
requirements and responsiveness of a developing embryo change as
the embryo passes various developmental milestones. Consequently,
accurate and timely knowledge of the developmental stage of an
embryonic culture would allow the skilled practitioner to
beneficially adjust the growth media components and other
environmental factors to achieve optimal embryo survival, growth,
and maturation. In addition, an understanding of developmentally
regulated genes would allow for early selection of advantageous
clones and provide tools for developmentally regulated transgenic
expression systems.
[0019] Currently, a reasonable determination of the precise
developmental stage of an embryo requires a practiced, physical
familiarity with the morphological appearance of embryos at
different stages, which is further complicated by the presence of
morphological variations between species. Consequently, visual
determination is performed best by experts in the field. Thus,
there is a need in the art for a staging method which can be
reliably practiced by the ordinary practitioner. The current
invention will allow one to stage embryos based on a relational
database system profiling gene expression patterns instead of
physical morphological differences, thereby permitting one less
skilled in the art of visual staging to biologically determine the
stages of embryogenesis.
[0020] The traditional morphological staging method provides only a
crude indication of the underlying biochemical condition or state
of an embryo. This level of information is insufficient for
refining culture conditions, including media formulations, or for
selecting potentially advantageous embryo clones for further
development. Thus, there is a need in the art for a more sensitive
staging method that precisely defines the physiological age,
health, growth requirements, and potential fitness of a particular
embryo. The current invention will allow definitive staging
significantly beyond that currently practiced in the art, and
provides a detailed analysis of the biochemical state and potential
fitness of an embryo by comparison to developed relational database
profiles.
[0021] Visual staging methods depend on morphological markers to
assign a numerical stage of 1-9 to an embryo. Nevertheless, it is
well accepted that visually undetectable developmental changes
occur in an embryo after it reaches stage 9. The current invention
is particularly useful in providing means for monitoring and
evaluating the developmental state of these older embryos, as
genetic responses occur and are detectable up to and through an
adult tree's life.
[0022] There further exists in the art a need for information
regarding the proteins, genes, and gene expression patterns in
plant embryo development, as well as a more thorough understanding
of how this information relates to the physiology, developmental
potential, and genetic quotient of a plant embryo. The relational
database system provides a platform for which to monitor individual
gene expression levels during embryo development while directly
correlating expression with, for example, environmental conditions,
age, and embryo fitness, as well as the protein identification
achieved by BLAST searches of publicly available databases (i.e.,
GenBank) for desirable genes. Accordingly, the present invention
therefore provides the additional ability to correlate the direct,
global gene expression response within the embryo system to a
typically non-expressing gene driven by a stage-specific
promoter.
SUMMARY OF THE INVENTION
[0023] The present invention addresses these needs by providing in
a relational database format nucleic acid and protein sequences
that are differentially expressed during various stages of plant
embryogenesis. The invention encompasses a set of isolated nucleic
acid molecules comprising the DNA sequence of any one of SEQ ID
NOS: 1-334 and nucleic acid molecules related or complementary to
any one of SEQ ID NOS: 1-334. (See Table I) As such, the invention
includes both single-stranded and double-stranded RNA and DNA
nucleic acids, including variants thereof. The nucleic acids of the
invention can be used as an expression template in the form of DNA
arrays, including for example, gene arrays, DNA chips, and dot
array Southerns, for which to compare and evaluate expression in
test samples. (See Table II) The nucleic acids of the invention can
be further used as probes to detect the presence or level of both
single-stranded and double-stranded RNA and DNA encoding variants
of polypeptides or fragments of polypeptides encompassed by the
invention. The nucleic acids of the invention can be further used
as promoters for the expression of sense and antisense molecules at
specific stages of embryo development. Data acquired through the
use of the present invention can in turn be provided to the
relational database for further development.
[0024] Isolated nucleic acid molecules that hybridize to a
denatured, double-stranded DNA comprising the DNA sequence of any
one of SEQ ID NOS: 1-334 under conditions of moderate or high
stringency are also encompassed by the invention. The invention
further encompasses synthetic and naturally-occurring variants of
the nucleic acids described in SEQ ID NOS: 1-334, for example,
isolated nucleic acid molecules derived by in vitro mutagenesis
from SEQ ID NOS: 1-334. In vitro mutagenesis would include numerous
techniques known in the art including, but not limited to,
site-directed mutagenesis, random mutagenesis, and in vitro nucleic
acid synthesis.
[0025] The invention also encompasses related molecules (variants)
including isolated nucleic acid molecules degenerate from SEQ ID
NOS: 1-334 as a result of the genetic code, for example,
naturally-occurring or synthetic allelic variants of the genes
encoding SEQ ID NOS: 1-334. Such related molecules also encompass
both smaller and larger nucleic acids that contain sufficient
sequence to support hybridization to any of SEQ ID NOS: 1-334 under
conditions of moderate or high stringency. Consequently,
recombinant vectors, including those that direct the expression of
these nucleic acid molecules and host cells transformed or
transfected with these vectors are herein defined as variants and
are encompassed by the invention.
[0026] Another embodiment of this invention is the production of
transgenic vectors and transgenic plants comprising vectors or
other nucleic acids comprising any one of SEQ ID NOS: 1-334 and
related molecules. Particularly preferred are those capable of
expressing polypeptides or peptides encoded by any of SEQ ID NOS:
1-327. In a preferred embodiment, the transgene comprises SEQ ID
NO: 327, or a variant thereof.
[0027] SEQ ID NO: 327 encodes a protein which corresponds to a
novel Loblolly pine homolog of the plant Major Intrinsic Protein
(MIP) family. MIPs comprise a large family of related proteins that
function as membrane channels for the transport of water and
possibly ions across cellular membranes. Henceforth, the encoded
protein of SEQ ID NO: 327 may be referred to as Loblolly MIP.
Variants, including naturally-occurring and
artifactually-programmed allelic variants, vectors, and other
nucleic acids which hybridize to SEQ ID NO: 327 under conditions of
moderate or high stringency are encompassed by the invention. Also
encompassed are plant cells, seeds, embryos and trees, transgenic
for loblolly pine MIP, and variants thereof.
[0028] The invention also encompasses isolated polypeptides, or
fragments thereof, encoded by any one of the nucleic acid molecules
of SEQ ID NOS: 1-327, including variants thereof. The invention
further encompasses the use of these peptide sequences as markers
for staging, monitoring, and selecting embryos and embryo cultures.
The invention also encompasses methods for the production of these
polypeptides or fragments thereof including culturing a host cell
under conditions promoting expression and recovering the
polypeptide or peptide from the culture medium. In particular, the
expression of polypeptides or peptides encoded by SEQ ID NOS: 1-327
in viral vectors, bacteria, yeast, plant, and animal cells is
encompassed by the invention. Isolated polyclonal or monoclonal
antibodies that bind to peptides encoded by SEQ ID NOS: 1-327 are
also encompassed by the invention.
[0029] Further encompassed by this invention are methods for using
the nucleic acid molecules of any one of SEQ ID NOS: 1-327 to
obtain full length cDNA and genomic sequences of the corresponding
genes, including cognate, homologous, or otherwise related genetic
sequences, which hybridize to any of SEQ ID NOS: 1-327 under
conditions of moderate or high stringency. Also provided by this
invention are oligonucleotides derived from any one of SEQ ID NOS:
1-334 that can be used as probes and/or as primers in PCR, RT-PCR,
and other assays to detect the presence or level of the nucleic
acids of SEQ ID NOS: 1-334 and related molecules.
[0030] The primers and other probes of the invention may be used to
monitor and characterize the development of plant embryos, in
particular, pine tree embryos. Characterization of embryonic gene
expression provides means for correlating gene expression with
current and potential plant phenotypes. Consequently, the present
invention encompasses means for monitoring and adjusting growth
conditions (see FIG. 6), as well as means for selecting genetically
superior embryonic clones for further propagation and expansion
(see FIG. 8). Thus, the present invention encompasses the use of
DNA or RNA probes derived from the nucleic acid molecules of SEQ ID
NOS: 1-334 in any form, e.g., in DNA arrays, and antibodies raised
against polypeptides or peptide fragments encoded by SEQ ID NOS:
1-327, to determine relative or absolute levels of expression of
the genes or proteins encoded by SEQ ID NOS: 1-327. In addition,
these nucleic acid and antibody probes may be used for staging,
monitoring, characterizing, or selecting plant embryos or embryo
cultures, particularly pine tree embryos.
[0031] The relational database of the present invention allows
expression information pertaining to embryo stages to be viewed as
sequence data generated in accordance with the present invention.
The invention includes a database for storing a plurality of
sequence records for which to correlate embryo stages to sequence
records. The method further involves providing an interface which
allows a user to select one or more expression categories contained
within the database.
[0032] The relational database is designed to include separate
parts or cells for information storage. One cell or part may
contain a gene expression database which contains nucleic acid
molecules of SEQ ID NOS: 1-327. Other cells or parts may contain
descriptive information pertaining to each nucleic acid molecules
of SEQ ID NOS: 1-327, additional sequence data related to the gene
expression database, protein encoded by nucleic acids disclosed
herein, similarity values to known proteins of other systems, and
to conditions under which expression data was obtained.
[0033] The database system described in the present invention will
allow identification or selection of particular genes of interest
for further use with DNA arrays. Identification or selection of
particular genes may include, for example, those related to
patterns of expression, those identified with homology to known
genes from other studies, and those sequences considered novel.
BRIEF DESCRIPTION OF THE DRAWINGS
[0034] FIG. 1 depicts differential display of loblolly pine zygotic
and somatic embryos at different stages of development.
[0035] FIG. 2 displays embryo gene expression observed by
high-density array Southern hybridization.
[0036] FIG. 3 provides a general schematic for gene regulation
studies arising from the cDNA cloning of genes expressed in
embryos.
[0037] FIG. 4 depicts graphical representation of hybridization of
`dehydrin` and LPZ216 cDNA probes to total RNA isolated from
zygotic embryos of loblolly pine.
[0038] FIG. 5 displays ABA concentration of loblolly pine
embryos.
[0039] FIG. 6 shows schematic of gene study for improved somatic
embryogenesis.
[0040] FIG. 7 shows detection of gene expression by high-density
array Southern hybridization for loblolly pine genotype 333 after
12 weeks on two maturation media.
[0041] FIG. 8 depicts the application of embryogenic gene
expression studies.
[0042] FIG. 9 displays slot blots and expression levels for three
embryogenesis-related genes.
[0043] FIG. 10 depicts clone LPS-097 sequence (LP2-3 differential
display fragment.)
[0044] FIG. 11 displays a northern blot for the LP2-3 gene during
stages 1-3.
[0045] FIG. 12 displays a slot blot of total RNA from somatic
embryo tissue probed with an LP2-3-specific probe.
[0046] FIG. 13 displays a slot blot of total RNA from zygotic
embryo tissue probed with an LP2-3-specific probe.
[0047] FIG. 14 depicts the quantified expression of early zygotic
embryos compared to early somatic embryos.
DETAILED DESCRIPTION OF THE INVENTION
[0048] The three hundred and twenty-seven differentially expressed
cDNAs isolated from plant specimens of known developmental ages are
disclosed in SEQ ID NOS: 1-327. The seven stage-specific promoters
isolated from plant specimens are disclosed in SEQ ID NOS: 328-334.
The discovery of these cDNAs and promoters enables the design,
isolation, and construction of related nucleic acids, proteins,
antigens, antibodies other heterologous genes. Both the cDNAs and
promoters facilitate the staging, characterization, and
manipulation of plant embyrogenesis, in particular, conifer
embryogenesis. These molecules, and related nucleic acids,
peptides, proteins, antigens, and antibodies are particularly
useful when compiled into a relational database for the monitoring,
design, selection, and cultivation of improved crop plants.
[0049] The cDNAs of SEQ ID NOS: 1-327, in addition to the promoters
of SEQ ID NOS: 328-334, were originally derived from Pinus taeda
embryos, commonly known as the Loblolly Pine. Nevertheless, it is
understood that the invention is applicable to and encompasses all
plants, including all dicotyledonous plants, including all
conifers, including all species of Pinus, Picea, and Pseudotsuga.
Exemplary conifers may include Picea abies, and Psedotsuga
menziesii, and Pinus taeda.
[0050] Nucleic Acid Molecules
[0051] In a particular embodiment, the invention relates to certain
isolated nucleotide sequences including those that are
substantially free from contaminating endogenous material. The
terms "nucleic acid" or "nucleic acid molecule" refer to a
deoxyribonucleotide or ribonucleotide polymer in either single-or
double-stranded form, and unless otherwise limited, would encompass
known analogs of natural nucleotides that can function in a similar
manner as naturally occurring nucleotides. A "nucleotide sequence"
also refers to a polynucleotide molecule or oligonucleotide
molecule in the form of a separate fragment or as a component of a
larger nucleic acid. The nucleotide sequence or molecule may also
be referred to as a "nucleotide probe." The nucleic acid molecules
of the invention are derived from DNA or RNA isolated at least once
in substantially pure form and in a quantity or concentration
enabling identification, manipulation, and recovery of its
component nucleotide sequence by standard biochemical methods.
Examples of such methods, including methods for PCR protocols that
may be used herein, are disclosed in Sambrook et al., Molecular
Cloning: A Laboratory Manual, 2nd ed., Cold Spring Harbor
Laboratory, Cold Spring Harbor, N.Y. (1989), Current Protocols in
Molecular Biology edited by F. A. Ausubel et al., John Wiley and
Sons, inc. (1987), and Innis, M. et al., eds., PCR Protocols: A
Guide to Methods and Applications, Academic Press (1990), each of
which are herein incorporated by reference in their entirety.
[0052] As used herein a "nucleotide probe" is defined as an
oligonucleotide capable of binding to a target nucleic acid of
complementary sequence through one or more types of chemical bonds,
through complementary base pairing, or through hydrogen bond
formation. As described above, the oligonucleotide probe may
include natural (ie. A, G, C, or T) or modified bases
(7-deazaguanosine, inosine, etc.). In addition, bases in a
oligonucleotide probe may be joined by a linkage other than a
phosphodiester bond, so long as it does not prevent hybridization.
Thus, oligonucleotide probes may have constituent bases joined by
peptide bonds rather than phosphodiester linkages.
[0053] A "target nucleic acid" herein refers to a nucleic acid to
which the nucleotide probe or molecule can specifically hybridize.
The probe is designed to determine the presence or absence of the
target nucleic acid, and the amount of target nucleic acid. The
target nucleic acid has a sequence that is complementary to the
nucleic acid sequence of the corresponding probe directed to the
target. As recognized by one of skill in the art, the probe may
also contain additional nucleic acids or other moieties, such as
labels, which may not specifically hybridize to the target. The
term target nucleic acid may refer to the specific nucleotide
sequence of a larger nucleic acid to which the probe is directed or
to the overall sequence (e.g., gene or mRNA) whose expression level
it is desired to detect. One skilled in the art will recognize the
full utility under various conditions.
[0054] As described herein, the nucleic acid molecules of the
invention include DNA in both single-stranded and double-stranded
form, as well as the RNA complement thereof. DNA includes, for
example, cDNA, genomic DNA, chemically synthesized DNA, DNA
amplified by PCR, and combinations thereof. Genomic DNA, including
translated, non-translated and control regions, may be isolated by
conventional techniques, e.g., using any one of the cDNAs of SEQ ID
NO: 1 through SEQ ID NO: 327, or suitable fragments thereof, as a
probe, to identify a piece of genomic DNA which can then be cloned
using methods commonly known in the art. In general, nucleic acid
molecules within the scope of the invention include sequences that
hybridize to sequences of SEQ ID NOS: 1-334 under hybridization and
wash conditions of 5.degree., 10.degree., 15.degree., 20.degree.,
25.degree., or 30.degree. below the melting temperature of the DNA
duplex of sequences of SEQ ID NOS: 1-334, including any range of
conditions subsumed within these ranges.
[0055] DNA Arrays
[0056] In a further embodiment, DNA arrays are used to identify
hybridizing sequences from test samples. The term "DNA array"
refers to "gene arrays," "DNA chips," "dot array Southerns," etc.
One of skill in the art will appreciate that an enormous number of
array designs are suitable for the practice of this invention. The
DNA array will typically include one or a multiplicity of nucleic
acid molecules derived from SEQ ID NO: 1 through SEQ ID NO: 327
that specifically hybridize to the nucleic acid expression of which
is to be detected. In addition, the array may include one or more
control probes to monitor the expression system. Control probes
refer to known expression products present at each stage of
expression, e.g., ribosomal gene products or the transcripts of
other housekeeping genes. The organization of the DNA array will be
known to facilitate interpretation of results. Examples in the art
describing the uses and composition of DNA arrays can be found in
U.S. Pat. Nos.: 5,700,637, 5,837,832, 5,843,655, 5,874,219,
6,040,138, 6,045,996, and are incorporated by reference.
[0057] Molecules That Hybridize to Identified Sequences
[0058] Thus, in a particular embodiment, this invention provides an
isolated nucleic acid molecule selected from the group consisting
of:
[0059] (1) a DNA sequence comprising any one of the sequences
presented in SEQ ID NO: 1 through SEQ ID NO: 334;
[0060] (2) an isolated nucleic acid molecule that hybridizes to
either strand of a denatured, double-stranded DNA comprising the
nucleic acid sequence of (a) under conditions of moderate
stringency; and
[0061] (3) an isolated nucleic acid molecule that hybridizes to
either strand of a denatured, double-stranded DNA comprising the
nucleic acid sequence of (a) under conditions of high
stringency.
[0062] As used herein, stringency conditions in nucleic acid
hybridizations can be readily determined by those having ordinary
skill in the art based on, for example, the length and composition
of the nucleic acid. In one embodiment, moderate stringency is
herein defined as a nucleic acid having 10, 11, 12, 13, 14, 15, 16,
or 17, contiguous nucleotides identical to any of the sequences of
SEQ ID NOS: 1-334, or a complement thereof. Similarly, high
stringency is hereby defined as a nucleic acid having 18, 19, 20,
21, 22, or more contiguous identical nucleotides, or a longer
nucleic acid having at least 80, 85, 90, 95, or 99 percent identity
with any of the sequences of SEQ ID NOS: 1-334; for sequences of at
least 50, 100, 150, 200, or 250 nucleotides, high stringency may
comprise an overall identity of at least 60, 65, 70 or 75
percent.
[0063] Generally, nucleic acid hybridization simply involves
providing a denatured nucleotide molecule or probe and target
nucleic acid under conditions where the probe and its complementary
target can form stable hybrid duplexes through complementary base
pairing. The nucleic acids that do not substantially form hybrid
duplexes are then washed away leaving the hybridized nucleic acids
to be detected, typically through detection of an attached
detectable label. It is further generally recognized that nucleic
acids are denatured by increasing the temperature or decreasing the
salt concentration of the buffer containing the nucleic acids.
Under lower stringency conditions (e.g., low temperature and/or
high salt) hybrid duplexes (e.g., DNA:DNA, RNA:RNA, or RNA:DNA)
will form even where the annealed sequences are not perfectly
complementary. Thus specificity of hybridization is reduced at
lower stringency. Conversely, at higher stringency (e.g., higher
temperature or lower salt) successful hybridization requires fewer
mismatches. One of skill in the art will appreciate that
hybridization conditions may be selected to provide any degree of
stringency.
[0064] As used herein, the percent identity between an amino acid
sequence encoded by any of SEQ ID NOS: 1-334 and a potential
hybridizing variant can be determined, for example, by comparing
sequence information using the GAP computer program, version 6.0
described by Devereux et al. (Nucl. Acids Res. 12:387, 1984) and
available from the University of Wisconsin Genetics Computer Group
(UWGCG). The GAP program utilizes the alignment method of Needleman
and Wunsch (J. Mol. Biol, 48:443, 1970), as revised by Smith and
Waterman (Adv. Appl. Math 2:482, 1981). The preferred default
parameters for the GAP program include: (1) a unary comparison
matrix (containing a value of 1 for identities and 0 for
non-identities) for nucleotides, and the weighted comparison matrix
of Gribskov and Burgess (Nuci. Acids Res. 14:6745, 1986), as
described by Schwartz and Dayhoff (eds., Atlas of Protein Sequence
and Structure, National Biomedical Research Foundation, pp.
353-358, 1979); (2) a penalty of 3.0 for each gap and an additional
0.10 penalty for each symbol in each gap; and (3) no penalty for
end gaps.
[0065] Alternatively, basic protocols for empirically determining
hybridization stringency are set forth in section 2.10 of Current
Protocols in Molecular Biology edited by F. A. Ausubel et al., John
Wiley and Sons, Inc. (1987). Stringency conditions can be
determined readily by the skilled artisan. An example of moderate
stringency hybridization conditions would be hybridization in
5.times.SSC, 5.times. Denhardt's Solution, 50% (w/v) formamide, and
1% SDS at 42.degree. C. with washing conditions of 0.2.times.SSC
and 0.1% SDS at 42.degree. C. An example of high stringency
conditions can be defined as hybridization conditions as above, and
with washing at approximately 68.degree. C., in 0.1.times.SSC, and
0.1% SDS. The skilled artisan will recognize that the temperature
and wash solution salt concentration can be adjusted as necessary
according to factors such as the length of the probe.
[0066] Due to the degeneracy of the genetic code wherein more than
one codon can encode the same amino acid, multiple DNA sequences
can code for the same polypeptide. Such variant DNA sequences can
result from genetic drift or artificial manipulation (e.g.,
occurring during PCR amplification or as the product of deliberate
mutagenesis of a native sequence). The present invention thus
encompasses any nucleic acid capable of encoding a protein derived
from SEQ ID NOS: 1-327, or variants thereof.
[0067] Deliberate mutagenesis of a native sequence can be carried
out using numerous techniques well known in the art. For example,
oligonucleotide-directed site-specific mutagenesis procedures can
be employed, particularly where it is desired to mutate a gene such
that predetermined restriction nucleotides or codons are altered by
substitution, deletion or insertion. Exemplary methods of making
such alterations are disclosed by Walder et al. (Gene 42:133,
1986); Bauer et al. (Gene 37:73, 1985); Craik (BioTechniques, Jan.
12-19, 1985); Smith et al. (Genetic Engineering: Principles and
Methods, Plenum Press, 1981); Kunkel (Proc. Natl. Acad. Sci. USA
82:488, 1985); Kunkel et al. (Methods in Enzymol. 154:367, 1987);
and U.S. Pat. Nos. 4,518,584 and 4,737,462, all of which are
incorporated by reference.
[0068] Thus, the invention further provides an isolated nucleic
acid molecule selected from the group comprising of (1), (2), and
(3) above and further consisting of:
[0069] (4) an isolated nucleic acid molecule degenerate from SEQ ID
NOS: 1-334 as a result of the genetic code; and
[0070] (5) an isolated nucleic acid molecule selected from the
group consisting of an allelic variants and species homologs of SEQ
ID NOS: 1-334.
[0071] Obtaining Full Length cDNAs
[0072] The cDNAs isolated and cloned through the differential
display procedure will often only represent a partial sequence
(generally the 3' end) of the mRNA from which it was derived due to
the nature of the arbitrary primer used in the differential display
PCR reaction. Consequently, the cDNA sequences of SEQ ID NOS: 1-327
provide powerful tools for obtaining the sequences of full-length
cDNAs. This can be accomplished by using a partial cDNA as a probe
to identify and isolate the full length cDNA from a population of
full length cDNAs or from a full length cDNA library. As is well
known in the art, similar procedures can be used to identify
corresponding genomic DNA sequences.
[0073] Alternatively, one can obtain the 5' sequence of a partial
cDNA by performing Rapid Amplification of cDNA Ends (RACE)
procedures such as those disclosed in Frohman, Methods in
Enzymology, 218:340-356 (1993) and Bertling et al., PCR Methods and
Applications 3:95-99 (1993) which are hereby incorporated by
reference. For example, Clonetech Laboratories, Inc. (Palo Alto,
Calif.) offers a SMAR.TM. cDNA product line that allows one to
generate high quality full length cDNAs and cDNA libraries.
SMART.TM. technology can also be used to perform RACE. One skilled
in the art will readily recognize that there are other equivalent
products and procedures for obtaining full length cDNAs. Full
length cDNAs may be sequenced and their sequences compared to
sequences in public databases to assess their identities and/or
homologies to other known sequences.
[0074] Cloned full length cDNAs can be used in the construction of
expression vectors for the production and purification of pine tree
polypeptides which contain the pine tree peptides encoded by the
cDNAs of any one of SEQ ID NOS: 1-327.
[0075] Oligonucleotide Primers for PCR Assays
[0076] In another embodiment, the present invention encompasses
oligonucleotide fragments derived from any one of SEQ ID NO: 1
through SEQ ID NO: 327 or from the reverse complement sequence of
any one of SEQ ID NO: 1 through SEQ ID NO: 327. Such
oligonucleotides would be useful as primers in the performance of
RT-PCR assays to detect, or even quantify, pine embryo
stage-specific transcripts. Such oligonucleotide primers will
generally comprise from 10 to 25 nucleotides substantially
complementary to the ends of the target sequence and may contain
additional non-complementary nucleotides, for example, nucleotides
that generate a restriction endonuclease site or cloning junction.
Programs useful in selecting PCR primers may be used to design the
oligonucleotides of this invention, but use of such programs is not
necessary. By way of example, the Wisconsin Package.TM. software
available from the Genetic Computer Group (Madison, Wis.) includes
a program called Prime that can aid in selecting primers from a
given template sequence. Protocols for the design and optimization
of PCR reactions are commonly known in the art and are described in
Saiki et al., Science 239:487 (1988); Recombinant DNA Methodology,
Wu et al., eds., Academic Press, Inc., San Diego (1989), pp.
189-196; and PCR Protocols: A Guide to Methods and Applications,
Innis et al., eds., Academic Press, Inc. (1990).
[0077] Antisense Nucleic Acid Molecules
[0078] Other useful fragments of the nucleic acids include
antisense or sense oligonucleotides comprising a single-stranded
nucleic acid sequence (either RNA or DNA) capable of binding to
target mRNA (sense) or DNA (antisense) sequences. Antisense or
sense oligonucleotides, according to the present invention,
comprise a fragment of DNA from any one of SEQ ID NO: 1 through SEQ
ID NO: 327. Such a fragment generally comprises at least about 14
nucleotides, preferably from about 14 To about 30 nucleotides. The
ability to derive an antisense or a sense oligonucleotide, based
upon a cDNA sequence encoding a given protein is described in, for
example, Stein and Cohen (Cancer Res. 48:2659, 1988) and van der
Krol et al. (Bio/Techniques 6:958, 1988).
[0079] Binding of antisense or sense oligonucleotides to target
nucleic acid sequences results in the formation of duplexes or
other nucleic acid complexes inimical to efficient production of
gene products. The antisense oligonucleotides thus may be used to
block expression of proteins or the function of RNA. Antisense or
sense oligonucleotides further comprise oligonucleotides having
modified sugar-phosphodiester backbones (or other sugar linkages,
such as those described in WO91/06629) and wherein such sugar
linkages are resistant to endogenous nucleases. Such
oligonucleotides with resistant sugar linkages are stable in vivo
(i.e., capable of resisting enzymatic degradation) but retain
sufficient sequence specificity to be able to bind to target
nucleotide sequences.
[0080] Other examples of sense or antisense oligonucleotides
include those oligonucleotides which are covalently linked to
organic moieties, such as those described in WO 90/10448, and other
moieties that increases affinity of the oligonucleotide for a
target nucleic acid sequence, such as poly-(L-lysine). Further
still, intercalating agents, such as ellipticine, and alkylating
agents or metal complexes may be attached to sense or antisense
oligonucleotides. Such modifications may modify binding
specificities of the antisense or sense oligonucleotide for the
target nucleotide sequence.
[0081] Antisense or sense oligonucleotides may be introduced into a
cell containing the target nucleic acid sequence by any gene
transfer method, including, for example, lipofection,
CaPO.sup.4-mediated DNA transfection, electroporation, or by using
gene transfer vectors such as Epstein-Barr virus or adenovirus.
[0082] Sense or antisense oligonucleotides also may be introduced
into a cell containing the target nucleotide sequence by formation
of a conjugate with a ligand binding molecule, as described in WO
91/04753. Suitable ligand binding molecules include, but are not
limited to, cell surface receptors, growth factors, other
cytokines, or other ligands that bind to cell surface receptors. In
one embodiment, conjugation of the ligand binding molecule does not
substantially interfere with the ability of the ligand binding
molecule to bind to its corresponding molecule or receptor, or
block entry of the sense or antisense oligonucleotide or its
conjugated version into the cell.
[0083] Alternatively, a sense or an antisense oligonucleotide may
be introduced into a cell containing the target nucleic acid
sequence by formation of an oligonucleotide-lipid complex, as
described in WO 90/10448. The sense or antisense
oligonucleotide-lipid complex is preferably dissociated within the
cell by an endogenous lipase.
[0084] Polypeptides Encoded by Differentially-Expressed cDNAs
[0085] The cDNAs of SEQ ID NOS: 1-327 can be translated into amino
acid sequences potentially corresponding to portions of
developmentally-regulated plant proteins. These amino acid
sequences can be identified from sequences listed in Table I,
below. The cDNAs encoding these predicted polypeptides are grouped
into early, middle, and late transcripts according to the staged
embryo population from which they were derived.
[0086] (See Table I)
[0087] Although the term "peptide" is generally understood to
reference synthetic sequences, or fragments of larger proteins, and
includes short amino acid sequences of between 2 and 10 amino
acids, whereas "polypeptide" refers to larger sequences and
full-length proteins, the terms are used interchangeably herein to
indicate that the invention applies to peptides and polypeptides of
any length and variants thereof. Moreover, the discovery of
presumptive open reading frames in SEQ ID NOS: 1-327, and the
ability to isolate additional cDNA sequence, enables the
construction of expression vectors comprising nucleic acid
sequences encoding those polypeptides. The cDNAs of the invention
also enable cells transfected or transformed with expression
vectors driving the expression of the encoded polypeptides and
antibodies reactive with the polypeptides.
[0088] In one embodiment, the invention provides for isolated
polypeptides, preferably, pine tree polypeptides. As used herein,
the term "polypeptides" refers to a genus of polypeptide or peptide
fragments that encompass the amino acid sequences identified from
Table I, as well as smaller fragments. Consequently, the invention
encompasses any polypeptide fragment comprising at least 5, 6, 7,
8, 9, 10, 11, 12, 13, 14, or 15 contiguous amino acids encoded by
the cDNAs of any of SEQ ID NOS: 1-327, or comprising at least 5, 6,
7, 8, 9, 10, 11, 12, 13, 14, or 15 contiguous amino acids of any of
amino acid sequence derived from Table I.
[0089] Alternatively, a polypeptide may be defined in terms of its
antigenic relatedness to any peptide encoded by SEQ ID NOS:-1-327.
Thus, in one embodiment a polypeptide within the scope of the
invention is defined as an amino acid sequence comprising a linear
or 3-dimensional epitope shared with any peptide encoded by the
cDNAs of SEQ ID NOS: 1-327. Alternatively, a polypeptide within the
scope of the invention is recognized by an antibody that
specifically recognizes any peptide encoded by SEQ ID NOS: 1-327.
Antibodies are defined to be specifically binding if they bind pine
tree polypeptides with a K.sub.a of greater than or equal to about
10.sup.7 M.sup.-, and preferably greater than or equal to 10.sup.8
M.sup.-1.
[0090] A polypeptide "variant" as referred to herein means a
polypeptide substantially homologous to a native polypeptide, but
which has an amino acid sequence different from that encoded by any
of SEQ ID NOS: 1-327 because of one or more deletions, insertions
or substitutions. The variant amino acid sequence preferably is at
least 80% identical to a native polypeptide amino acid sequence,
preferably at least 90%, more preferably, at least 95% identical
over at least 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19,
20, 21-25, or 26-30 contiguous amino acids. The percent identity
between an amino acid sequence encoded by any of SEQ ID NOS: 1-327
and a potential variant can be determined manually, or, for
example, by comparing sequence information using the GAP computer
program, version 6.0 described by Devereux et al. (Nucl. Acids Res.
12:387, 1984) and available from the University of Wisconsin
Genetics Computer Group (UWGCG). The GAP program, described above,
utilizes the alignment method of Needleman and Wunsch (J. Mol.
Biol. 48:443,1970), as revised by Smith and Waterman (Adv. Appl.
Math 2:482, 1981).
[0091] Variants can comprise conservatively substituted sequences,
meaning that a given amino acid residue is replaced by a residue
having similar physiochemical characteristics. Examples of
conservative substitutions include substitution of one aliphatic
residue for another, such as lie, Val, Leu, or Ala for one another,
or substitutions of one polar residue for another, such as between
Lys and Arg; Glu and Asp; or Gin and Asn. See Zubay, Biochemistry,
Addison-Wesley Pub. Co., (1983) incorporated by reference in its
entirety. The effects of such substitutions can be calculated using
substitution score matrices such a PAM-120, PAM-200, and PAM-250 as
discussed in Altschul, (J. Mol. Biol. 219:555-65, 1991). Other such
conservative substitutions, for example, substitutions of entire
regions having similar hydrophobicity characteristics, are well
known.
[0092] Naturally-occurring peptide variants are also encompassed by
the invention. Examples of such variants are proteins that result
from alternate mRNA splicing events or from proteolytic cleavage of
the polypeptides of Table I. Variations attributable to proteolysis
include, for example, differences in the N- or C-termini upon
expression in different types of host cells, due to proteolytic
removal of one or more terminal amino acids from the polypeptides
encoded by the sequences of Table I (generally from 1-5 terminal
amino acids).
[0093] As stated above, the invention provides recombinant and
non-recombinant, isolated and purified polypeptides, preferably
pine tree polypeptides. Variants and derivatives of native
polypeptides can be obtained by isolating naturally-occurring
variants, or the nucleotide sequence of variants, of other plant
lines or species, or by artificially programming mutations of
nucleotide sequences coding for native pine tree polypeptides.
Alterations of the native amino acid sequence can be accomplished
by any of a number of conventional methods. Mutations can be
introduced at particular loci by synthesizing oligonucleotides
containing a mutant sequence, flanked by restriction sites enabling
ligation to fragments of the native sequence. Following ligation,
the resulting reconstructed sequence encodes an analog having the
desired amino acid insertion, substitution, or deletion.
Alternatively, oligonucleotide-directed site-specific mutagenesis
procedures can be employed to provide an altered gene wherein
predetermined codons can be altered by substitution, deletion or
insertion. Exemplary methods of making such alterations are
discussed supra.
[0094] The following sections are examples of the various
expression vectors, host cells, and protein purification methods
that are known in the art. These examples are provided merely as
illustrative and should not be construed as the only means to
express and purify the polypeptides and polypeptide variants of the
invention.
[0095] Expression Vectors and Purified proteins
[0096] Recombinant expression vectors containing a nucleic acid
sequence encoding the polypeptides of the invention can be prepared
using well known methods. In one embodiment, the expression vectors
include a cDNA sequence encoding the polypeptide operably linked to
suitable transcriptional or translational regulatory nucleotide
sequences, such as those derived from a mammalian, microbial,
viral, or insect gene. Examples of regulatory sequences include
transcriptional promoters, operators, or enhancers, mRNA ribosomal
binding sites, and appropriate sequences which control
transcription and translation initiation and termination.
Nucleotide sequences are "operably linked" when the regulatory
sequence functionally relates to the cDNA sequence of the
invention. Thus, a promoter nucleotide sequence is operably linked
to a cDNA sequence if the promoter nucleotide sequence controls the
transcription of the cDNA sequence. The ability to replicate in the
desired host cells, usually conferred by an origin of replication,
and a selection gene by which transformants are identified can
additionally be incorporated into the expression vector.
[0097] In addition, sequences encoding appropriate signal peptides
that are not naturally associated with the polypeptides of the
invention can be incorporated into expression vectors. For example,
a DNA sequence for a signal peptide (secretory leader) can be fused
in-frame to the pine tree nucleotide sequence so that the
polypeptides of the invention is initially translated as a fusion
protein comprising the signal peptide. A signal peptide that is
functional in the intended host cells enhances extracellular
secretion of the expressed polypeptide. The signal peptide can be
cleaved from the polypeptide upon secretion from the cell.
[0098] Fusions of additional peptide sequences at the amino and
carboxyl terminal ends of the polypeptides of the invention can be
used to enhance expression of the polypeptides or aid in the
purification of the protein. Such peptides include, for example,
poly-His or the antigenic identification peptides described in U.S.
Pat. No. 5,011,912 and in Hopp et al., (Bio/Technology6:1204,
1988).
[0099] Suitable host cells for expression of polypeptides of the
invention include prokaryotes, yeast or higher eukaryotic cells.
Appropriate cloning and expression vectors for use with bacterial,
fungal, yeast, and mammalian cellular hosts are described, for
example, in Pouwels et al., Cloning Vectors: A Laboratory Manual,
Elsevier, N.Y., (1985). Cell-free translation systems could also be
employed to the disclosed polypeptides using RNAs derived from DNA
constructs disclosed herein.
[0100] Prokaryotic Expression Systems
[0101] Prokaryotes include gram negative or gram positive
organisms, for example, E. coli or Bacilli. Suitable prokaryotic
host cells for transformation include, for example, E. coli,
Bacillus subtilis, Salmonella typhimurium, and various other
species within the genera Pseudomonas, Streptomyces, and
Staphylococcus. In a prokaryotic host cell, such as E. coli, the
disclosed polypeptides can include an N-terminal methionine residue
to facilitate expression of the recombinant polypeptide in the
prokaryotic host cell. The N-terminal methionine can be cleaved
from the expressed recombinant polypeptide.
[0102] Expression vectors for use in prokaryotic host cells
generally comprise one or more phenotypic selectable marker genes.
A phenotypic selectable marker gene is, for example, a gene
encoding a protein that confers antibiotic resistance or that
supplies an autotrophic requirement. Examples of useful expression
vectors for prokaryotic host cells include those derived from
commercially available plasmids such as the cloning vector pBR322
(ATCC 37017). pBR322 contains genes for ampicillin and tetracycline
resistance and thus provides simple means for identifying
transformed cells. To construct an expression vector using pBR322,
an appropriate promoter and a DNA sequence encoding one or more of
the polypeptides of the invention are inserted into the pBR322
vector. Other commercially available vectors include, for example,
pKK223-3 (Pharmacia Fine Chemicals, Uppsala, Sweden) and pGEM-1
(Promega Biotec, Madison, Wis., USA). Other commercially available
vectors include those that are specifically designed for the
expression of proteins; these would include pMAL-p2 and pMAL-c2
vectors that are used for the expression of proteins fused to
maltose binding protein (New England Biolabs, Beverly, Mass.,
USA).
[0103] Promoter sequences commonly used for recombinant prokaryotic
host cell expression vectors include P-lactamase (penicillinase),
lactose promoter system (Chang et al., Nature 275:615, 1978; and
Goeddel et al., Nature 281:544, 1979), tryptophan (trp) promoter
system (Goeddel et al., Nucl. Acids Res. 8:4057, 1980; and
EP-A-36776), and tac promoter (Maniatis, Molecular Cloning: A
Laboratory Manual, Cold Spring Harbor Laboratory, p. 412, 1982). A
particularly useful prokaryotic host cell expression system employs
a phage .lambda. P.sub.L promoter and a c1857ts thermolabile
repressor sequence. Plasmid vectors available from the American
Type Culture Collection ("ATCC"), which incorporate derivatives of
the PL promoter, include plasmid pHUB2 (resident in E. Coli strain
JMB9 (ATCC 37092)) and pPLc28 (resident in E. coli RR1 (ATCC
53082)).
[0104] DNA encoding one or more of the polypeptides of the
invention may be cloned in-frame into the multiple cloning site of
an ordinary bacterial expression vector. Ideally the vector would
contain an inducible promoter upstream of the cloning site, such
that addition of an inducer leads to high-level production of the
recombinant protein at a time of the investigator's choosing. For
some proteins, expression levels may be boosted by incorporation of
codons encoding a fusion partner (such as hexahistidine) between
the promoter and the gene of interest. The resulting "expression
plasmid" may be propagated in a variety of strains of E. coli.
[0105] For expression of the recombinant protein, the bacterial
cells are propagated in growth medium until reaching a
pre-determined optical density. Expression of the recombinant
protein is then induced, e.g., by addition of IPTG
(isopropyl-b-D-thiogalactopyranoside), which activates expression
of proteins from plasmids containing a lac operator/promoter. After
induction (typically for 1-4 hours), the cells are harvested by
pelleting in a centrifuge, e.g., at 5,000.times.G for 20 minutes at
4.degree. C.
[0106] For recovery of the expressed protein, the pelleted cells
may be resuspended in ten volumes of 50 mM Tris-HCl (pH 8)/1 M NaCl
and then passed two or three times through a French press. Most
highly expressed recombinant proteins forms insoluble aggregates
known as inclusion bodies. Inclusion bodies can be purified away
from the soluble proteins by pelleting in a centrifuge at
5,000.times.G for 20 minutes, 4.degree. C. The inclusion body
pellet is washed with 50 mM Tris-HCl (pH 8)/1% Triton X-100 and
then dissolved in 50 mM Tris-HCl (pH 8)/8 M urea/0.1 M DTT. Any
material that cannot be dissolved in 50 mM Tris-HCl (pH 8)/8 M
urea/0.1 M DTT may be removed by centrifugation (10,000.times.G for
20 minutes, 20.degree. C.). The protein of interest will, in most
cases, be the most abundant protein in the resulting clarified
supernatant. This protein may be "refolded" into the active
conformation by dialysis against 50 mM Tris-HCl (pH 8)/5 mM
CaCl.sub.2/5 mM Zn(OAc).sub.2/1 mM GSSG/0.1 mM GSH. After
refolding, purification can be carried out by a variety of
chromatographic methods such as ion exchange or gel filtration. In
some protocols, initial purification may be carried out before
refolding. As an example, hexahistidine-tagged fusion proteins may
be partially purified on immobilized Nickel.
[0107] While the preceding purification and refolding procedure
assumes that the protein is best recovered from inclusion bodies,
those skilled in the art of protein purification will appreciate
that many recombinant proteins are best purified out of the soluble
fraction of cell lysates. In these cases, refolding is often not
required, and purification by standard chromatographic methods can
be carried out directly.
[0108] Yeast Expression Systems
[0109] Polypeptides of the invention can also be expressed in yeast
host cells, preferably from the Saccharomyces genus (e.g., S.
cerevisiae). Other genera of yeast, such as Pichia or Kluyveromyces
(e.g. K. lactis), can also be employed. Yeast vectors will often
contain an origin of replication sequence from a 2.mu. yeast
plasmid, an autonomously replicating sequence (ARS), a promoter
region, sequences for polyadenylation, sequences for transcription
termination, and a selectable marker gene. Suitable promoter
sequences for yeast vectors include, among others, promoters for
metallothionine, 3-phosphoglycerate kinase (Hitzeman et al., J.
Biol. Chem. 255:2073, 1980), or other glycolytic enzymes (Hess et
al., J. Adv. Enzyme Reg. 7:149, 1968; and Holland et al., Biochem.
17:4900, 1978), such as enolase, glyceraldehyde-3-phosphate
dehydrogenase, hexokinase, pyruvate decarboxylase,
phosphofructokinase, glucose-6-phosphate isomerase,
3-phosphoglycerate mutase, pyruvate kinase, triosephosphate
isomerase, phosphoglucose isomerase, and glucokinase. Other
suitable vectors and promoters for use in yeast expression are
further described in Hitzeman, EPA-73,657 or in Fleer et. al.,
Gene, 107:285-195 (1991); and van den Berg et. al., Bio/Technology,
8:135-139 (1990). Another alternative is the glucose-repressible
ADH2 promoter described by Russell et al. (J. Biol. Chem. 258:2674,
1982) and Beier et al. (Nature 300:724, 1982). Shuttle vectors
replicable in both yeast and E. coli can be constructed by
inserting DNA sequences from pBR322 for selection and replication
in E. coli (Amp gene and origin of replication) into the
above-described yeast vectors.
[0110] The yeast .alpha.-factor leader sequence can be employed to
direct secretion of one or more of the disclosed polypeptides. The
.alpha.-factor leader sequence is often inserted between the
promoter sequence and the structural gene sequence. See, e.g.,
Kurjan et al., Cell 30:933, 1982; Bitter et al., Proc. Natl. Acad.
Sci. USA 81:5330, 1984; U.S. Pat. No. 4,546,082; and EP 324,274.
Other leader sequences suitable for facilitating secretion of
recombinant polypeptides from yeast hosts are known to those of
skill in the art. A leader sequence can be modified near its 3' end
to contain one or more restriction sites. This will facilitate
fusion of the leader sequence to the structural gene.
[0111] Yeast transformation protocols are known to those of skill
in the art. One such protocol is described by Hinnen et al., Proc.
Natl. Acad. Sci. USA 75:1929, 1978. The Hinnen et al. protocol
selects for Trp+transformants in a selective medium, wherein the
selective medium consists of 0.67% yeast nitrogen base, 0.5%
casamino acids, 2% glucose, 10 .mu.g/ml adenine, and 20 .mu.g/ml
uracil.
[0112] Yeast host cells transformed by vectors containing ADH2
promoter sequence can be grown for inducing expression in a "rich"
medium. An example of a rich medium is one consisting of 1% yeast
extract, 2% peptone, and 1% glucose supplemented with 80 .mu.g/ml
adenine and 80 .mu.g/ml uracil. Derepression of the ADH2 promoter
occurs when glucose is exhausted from the medium.
[0113] Mammalian Expression Systems
[0114] Mammalian or insect host cell culture systems could also be
employed to express recombinant polypeptides of the invention.
Baculovirus systems for production of heterologous proteins in
insect cells are reviewed by Luckow and Summers, Bio/Technology
6:47 (1988). Established cell lines of mammalian origin also can be
employed. Examples of suitable mammalian host cell lines include
the COS-7 line of monkey kidney cells (ATCC CRL 1651) (Gluzman et
al., Cell 23:175, 1981), L cells, C127 cells, 3T3 cells (ATCC CCL
163), Chinese hamster ovary (CHO) cells, HeLa cells, and BHK (ATCC
CRL 10) cell lines, and the CV-1/EBNA-1 cell line (ATCC CRL 10478)
derived from the African green monkey kidney cell line CVI (ATCC
CCL 70) as described by McMahan et al. (EMBO J. 10: 2821,
1991).
[0115] Established methods for introducing DNA into mammalian cells
have been described (Kaufman, R. J., Large Scale Mammalian Cell
Culture, 1990, pp. 15-69). Additional protocols using commercially
available reagents, such as Lipofectamine (Gibco/BRL) or
Lipofectamine-Plus, can be used to transfect cells (Feigner et al.,
Proc. Natl. Acad. Sci. USA 84:7413-7417, 1987). In addition,
electroporation can be used to transfect mammalian cells using
conventional procedures, such as those in Sambrook et al. Molecular
Cloning: A Laboratory Manual, 2 ed. Vol. 1-3, Cold Spring Harbor
Laboratory Press, 1989). Selection of stable transformants can be
performed using resistance to cytotoxic drugs as a selection
method. Kaufman et al., Meth. in Enzymology 185:487-511, 1990,
describes several selection schemes, such as dihydrofolate
reductase (DHFR) resistance. A suitable host strain for DHFR
selection can be CHO strain DX-B11, which is deficient in DHFR
(Urlaub and Chasin, Proc. Natl. Acad. Sci. USA 77:4216-4220, 1980).
A plasmid expressing the DHFR cDNA can be introduced into strain
DX-B11, and only cells that contain the plasmid can grow in the
appropriate selective media. Other examples of selectable markers
that can be incorporated into an expression vector include cDNAs
conferring resistance to antibiotcs, such as G418 and hygromycin B.
Cells harboring the vector can be selected on the basis of
resistance to these compounds.
[0116] Transcriptional and translational control sequences for
mammalian host cell expression vectors can be excised from viral
genomes. Commonly used promoter sequences and enhancer sequences
are derived from polyoma virus, adenovirus 2, simian virus 40
(SV40), and human cytomegalovirus. DNA sequences derived from the
SV40 viral genome, for example, SV40 origin, early and later
promoter, enhancer, splice, and polyadenylation sites can be used
to provide other genetic elements for expression of a structural
gene sequence in a mammalian host cell. Viral early and late
promoters are particularly useful because both are easily obtained
from a viral genome as a fragment, which can also contain a viral
origin of replication (Fiers et al., Nature 273:113, 1978; Kaufman,
Meth. in Enzymology, 1990). Smaller or larger SV40 fragments can
also be used, provided the approximately 250 bp sequence extending
from the Hind III site toward the Bgl I site located in the SV40
viral origin of replication site is included.
[0117] Additional control sequences shown to improve expression of
heterologous genes from mammalian expression vectors include such
elements as the expression augmenting sequence element (EASE)
derived from CHO cells (Morris et al., Animal Cell Technology,
1997, pp. 529-534) and the tripartite leader (TPL) and VA gene RNAs
from Adenovirus 2 (Gingeras et al., J. Biol. Chem.
257:13475-13491,1982). The internal ribosome entry site (IRES)
sequences of viral origin allows dicistronic mRNAs to be translated
efficiently (Oh and Sarnow, Current Opinion in Genetics and
Development 3:295-300, 1993; Ramesh et al., Nucleic Acids Research
24:2697-2700, 1996). Expression of a heterologous cDNA as part of a
dicistronic mRNA followed by the gene for a selectable marker (eg.
DHFR) has been shown to improve transfectability of the host and
expression of the heterologous cDNA (Kaufman, Meth. in Enzymology,
1990). Exemplary expression vectors that employ dicistronic mRNAs
are pTR-DC/GFP described by Mosser et al., Biotechniques
22:150-161, 1997, and p2A51 described by Morris et al., Animal Cell
Technology, 1997, pp. 529-534.
[0118] A useful high expression vector, pCAVNOT, has been described
by Mosley et al., Cell 59:335-348,1989. Other expression vectors
for use in mammalian host cells can be constructed as disclosed by
Okayama and Berg (Mol. Cell. Biol. 3:280, 1983). A useful system
for stable high level expression of mammalian cDNAs in C127 murine
mammary epithelial cells can be constructed substantially as
described by Cosman et al. (Mol. Immunol. 23:935, 1986). A useful
high expression vector, PMLSV N1/N4, described by Cosman et al.,
Nature 312:768, 1984, has been deposited as ATCC 39890. Additional
useful mammalian expression vectors are described in EP-A-0367566,
and in U.S. patent application Ser. No. 07/701,415, filed May 16,
1991, incorporated by reference herein. The vectors can be derived
from retroviruses. In place of the native signal sequence, a
heterologous signal sequence can be added, such as the signal
sequence for IL-7 described in U.S. Pat. No. 4,965,195; the signal
sequence for IL-2 receptor described in Cosman et al., Nature
312:768 (1984); the IL4 signal peptide described in EP 367,566; the
type I IL-1 receptor signal peptide described in U.S. Pat. No.
4,968,607; and the type H IL-1 receptor signal peptide described in
EP 460,846.
[0119] The polypeptides of the invention and the nucleic acid
molecules encoding them can also be used as reagents to identify
(a) proteins that the disclosed polypeptides or their constituent
proteins regulate, and (b) other proteins with which it might
interact. The disclosed polypeptides can be coupled to a
recombinant protein, to an affinity matrix, or by using them as a
bait in the yeast two-hybrid system. The use of the yeast
two-hybrid system developed by Stanley Fields and coworkers is well
known in the art and described in Golemis, E., et al Section 20.1
in: Current Protocols in Molecular Biology, ed. Ausubel, F. M., et
al., John Wiley & Sons, NY, 1997 and in The Yeast Two-Hybrid
System., ed. P. L. Bartel and S. Fields, Oxford University Press,
1997.
[0120] Antibodies and Peptide Binding Proteins
[0121] Purified polypeptides of the invention can be used to
generate antibodies that bind to one or more epitopes of the
disclosed polypeptide. Such anti-polypeptide antibodies includes
polyclonal antibodies, monoclonal antibodies, fragments thereof
such as F(ab').sub.2, and Fab fragments, as well as any
recombinantly produced binding partners. Antibodies are defined to
be specifically binding if they bind pine tree polypeptides with a
K.sub.a of greater than or equal to about 10.sup.7 M.sup.-1.
Affinities of binding partners or antibodies can be readily
determined using conventional techniques, for example, those
described by Scatchard et al., Ann. N.Y. Acad. Sci., 51:660
(1949).
[0122] Polyclonal antibodies can be readily generated from a
variety of sources, for example, horses, cows, goats, sheep, dogs,
chickens, rabbits, mice, hamsters, guinea pigs, or rats, using
procedures that are well-known in the art, for example, as
described for example, U.S. Pat. No. 5,585,100, incorporated by
reference herein. In general, a composition comprising at least one
of the polypeptides of the invention is administered to the host
animal, typically through intra-peritoneal or subcutaneous
injection. In the case where a peptide is used as the immunogen, it
is preferable to conjugated it to a suitable carrier molecule, such
as a T-dependent antigen (Bovine Serum Albumin, cholera toxin, and
the like). The immunogenicity of the disclosed polypeptides can
also be enhanced through the use of an adjuvant, for example,
Freund's complete or incomplete adjuvant or alum. Following booster
immunizations, small samples of serum are collected and tested for
reactivity to the disclosed polypeptides or their constituent
epitopes. Examples of various assays useful for such determination
include those described in: Antibodies: A Laboratory Manual, Harlow
and Lane (eds.), Cold Spring Harbor Laboratory Press, 1988; as well
as procedures such as countercurrent immuno-electrophoresis (CIEP),
radioimmunoassay, radio-immunoprecipitation, enzyme-linked
immuno-sorbent assays (ELISA), dot blot assays, and sandwich
assays, see U.S. Pat. Nos. 4,376,110 and 4,486,530, each of which
is incorporated by reference in their entirety.
[0123] Monoclonal antibodies (or fragments thereof), directed
against epitopes of the disclosed polypeptides can also be readily
prepared using well-known procedures, such as, for example, the
procedures described in U.S. Patent No. RE 32,011, U.S. Pat. Nos.
4,902,614, 4,543,439, and 4,411,993; Monoclonal Antibodies,
Hybrddomas: A New Dimension in Biological Analyses, Plenum Press,
Kennett, McKearn, and Bechtol (eds.), 1980, each of which is
incorporated by reference. Briefly, the host animals, such as mice,
are injected intraperitoneally at least once, and preferably at
least twice at about 3 week intervals with isolated and purified
polypeptides optionally in the presence of adjuvant. Again, if
peptide fragments are used they may need to be conjugated to a
suitable carrier protein. Mouse sera are then assayed by
conventional dot blot technique or antibody capture (ABC) to
determine which animal is best to fuse. Approximately two to three
weeks later, the mice are given an intravenous boost of pine tree
polypeptides. Mice are later sacrificed and spleen cells fused with
commercially available myeloma cells, such as Ag8.653 (ATCC),
following established protocols. Briefly, the myeloma cells are
washed several times in media and fused to mouse spleen cells at a
ratio of about three spleen cells to one myeloma cell. The fusing
agent can be any suitable agent used in the art, for example,
polyethylene glycol (PEG). Fusion is plated out into plates
containing media that allows for the selective growth of the fused
cells. The fused cells can then be allowed to grow for
approximately eight days. Supernatants from resultant hybridomas
are collected and added to a plate that is first coated with goat
anti-mouse Ig. Following washes, a label, such as, .sup.125I-pine
tree polypeptides is added to each well followed by incubation.
Positive wells can be subsequently detected by autoradiography.
Positive clones can be grown in bulk culture and supernatants are
subsequently purified over a Protein A column (Pharmacia).
[0124] Monoclonal antibodies and specific-binding fragments of the
invention can be produced using alternative techniques, such as
those described by Alting-Mees et al., "Monoclonal Antibody
Expression Libraries: A Rapid Alternative to Hybridomas",
Strategies in Molecular Biology 3:1-9 (1990), which is incorporated
herein by reference. Similarly, binding partners can be constructed
using recombinant DNA techniques to incorporate the variable
regions of a gene that encodes a specific binding antibody Such a
technique is described in Larrick et al., Biotechnology, 7:394
(1989).
[0125] It is understood of course that many techniques could be
used to generate antibodies against the polypeptides of the
invention and that the above embodiments in no way limits the scope
of the invention.
[0126] Nucleotides. Proteins, Antibodies, and Binding Proteins As
Probes and Reagents
[0127] The disclosed nucleic acids, polypeptides, and antibodies
directed against the disclosed polypeptides can be used in a
variety of research protocols, such as in DNA arrays or as
reagents. A sample of such research protocols are given in Sambrook
et al. Molecular Cloning: A Laboratory Manual, 2 ed. Vol. 1-3, Cold
Spring Harbor Laboratory Press, (1989), incorporated by reference.
For example, the compiled sequences, polypeptides, etc., can serve
as markers for cell specific or tissue specific expression of RNA
or proteins. Similarly, this system can be used to investigate
constitutive and transient expression of the genes encoding the
cDNAs of SEQ ID NOS: 1-327 and the proteins encoded by these
genes.
[0128] Further, the disclosed cDNA sequences can be used to
determine the chromosomal location of the genomic DNA and to map
genes in relation to this chromosomal location. The disclosed
nucleotide sequence can be further used to identify additional
genes related to the nucleotides of SEQ ID NOS: 1-334 and to
establish evolutionary relatedness among species based on the
comparison of sequences. The disclosed nucleotide and polypeptide
sequences can be used to select for those genes or proteins that
are homologous to the disclosed cDNAs or polypeptides, using
well-established positive screening procedures such as Southern
blotting and immunoblotting and negative screening procedures such
as subtractive hybridization.
[0129] Method for Using Nucleic Acid Probes or Antibodies to Stage
Embryos
[0130] Accurate staging of tree embryos is critical. It is known
that different stages of tree embryos have different capacities as
subjects for genetic transformation and genetic engineering. In
addition, environmental requirements exhibited by embryos vary due
to increasing physiologic age. Currently, the staging of tree
embryogenesis is most accurately performed by an expert in the
field who is very familiar with the morphological appearance of
embryos at different stages. The cDNAs and related molecules of
this invention can be used as markers for different stages of tree
embryogenesis, thereby eliminating the need for a subjective eye to
assess maturity and potentially allowing for more accurate staging
of tree embryos. Moreover, by monitoring the expression of the
underlying genes, it is possible to determine when an embryo has
reached a certain level of development even if that level does not
correspond to a visible difference in embryo morphology. The
relational database of this invention aids the ability to monitor
expression levels and tailor research approaches, such as the use
of DNA arrays, to the specific needs of the objective, i.e.,
staging.
[0131] The information provided in this invention can be used in
whole or in part to stage embryos. For example, one or a
multiplicity of nucleic acid molecules from SEQ ID NOS: 1-327
having an expression profile consistent with a particular embryo
stage can be used in this invention. A researcher may find it
beneficial to use oligonucleotide probes or antibodies, for
example, that specifically recognize proteins derived from genes
expressed during middle embryonic stages, or that specifically
monitor expression levels for embryos that have reached maturity
associated with late developmental stages. A researcher can quickly
determine that an embryo subset has progressed to or through an
embryonic stage with the use of this invention and make appropriate
changes in conditions if necessary, e.g. alter growth media or
other environmental conditions.
[0132] Method for Monitoring, Enhancing, or Determining Expression
of Stage-Specific Genes
[0133] Expression patterns of SEQ ID NOS: 1-327 indicate that gene
activation can be classified as stage-specific, such as in the case
of SEQ ID NO: 327, otherwise known as "LP2-3." The promoter that
drives such a gene can perform valuable functions. For example, a
promoter from LP2-3 operatively linked to a reporter gene presented
within an embryo system is expected to produce the reporter product
under the conditions for expression of gene LP2-3. Thus, the system
allows a rapid determination of stage specific embryos by a simple
phenotypic reporter screen, perhaps by visualization of green
fluorescent protein (GFP) or by loss of fluorescent protein
product. Similarly, a set of promoters from known, differently
staged genes operatively linked to reporter genes will be effective
for monitoring developmental changes within the system as the
embryos mature. The LP2-3 promoter is identified as SEQ ID NOS:
328-334 in Table I. The promoter expression pattern is that of the
natively linked gene, LP2-3.
[0134] Virtually any indicator or reporter gene can be used for
this approach or for other methods associated with this invention
provided they are compatible with the system studied. Generally,
reporter genes are genes typically not present in the recipient
organism or tissue and which encode for proteins resulting in some
phenotypic change or enzymatic property. Examples of such genes and
assays are provided by Schenborn, E. and Groskreutz, D., Mol.
Biotechnol., 13:29, 1999; Helfand, S. L. and Rogina, B., Results
Probl. Cell Differ., 29:67, 2000; Kricka, L. J., Methods Enzymol.,
305:333, 2000; Himes, S. R. and Shannon, M. F., Methods Mol. Biol.,
130:165, 2000; and Leffel, S. M. et al., Biotechniques, 23:912,
1997, which are incorporated in their entirety by reference. In one
embodiment of this invention, the reporter used is GFP, or any
ariant of the fluorescent protein.
[0135] Additionally, one skilled in the art would recognize that a
promoter, like that from LP2-3, has potential to stimulate
production of products not ordinarily observed at a particular
stage. A promoter derived from a gene that expresses during a known
stage, for example an early stage, can be operatively linked to a
gene that does not normally express during that stage, yielding
controlled expression of any targeted gene. It may be shown that
earlier or later expression, or prolonged expression of a
particular gene may give a desirable genotype or phenotype in a
mature plant, may result in increased vigor in culture, or may be
sufficient to alter the normal maturation process of the embryo.
Prolonged expression of any desired gene also may be achieved from
linking a constitutively expressed promoter to the targeted gene.
Further, the ability to manipulate gene expression during
embryogenesis allows for a detailed study of the effects of an
individual gene or multiple genes on embryogenesis, leading to a
better understanding of the developmental processes involved in
embryogenesis.
[0136] Method of Correlatinq Gene Expression with Improved Tree
Stock or Culture Conditions
[0137] Importantly, the cDNAs and related molecules of the
invention can also be used as markers to examine genetic
heterogeneity and heredity through the use of techniques such as
genetic fingerprinting. These markers can also be correlated with
improved agronomic traits including good initiation frequency,
embryonic maturation, high frequency of germination, rapid growth
rates, herbicide tolerance, insect resistance, pathogen resistance,
climate and environmental adaptability wood quality, and wood fiber
quality and content, to name a few. Additionally, the expression of
these developmentally regulated genes can be compared among
genetically identical clones grown under different culture
conditions to determine the best protocols and media for somatic
embryogenesis.
[0138] Cryogenic storage of pine tree embryos is effective for
maintaining stocks of embryos determined by this invention to have
the desired fitness traits or exist at the appropriate
developmental stage. With such storage, one can specifically target
desirable embryos for expansion many years after they are frozen.
For example, a culture of somatic embryos can be divided into at
least three portions, one of which is cryogenically stored, one
which is used to study gene embryonic gene (and protein)
expression, and one that is used to produce seedlings for field
testing. Clones producing valuable mature plants could be selected
and expanded from frozen stocks. Additional clones exhibiting
similar expression patterns could be selected for future expansion
and cultivation.
[0139] As will be evident to the ordinary practitioner, there are
numerous ways in which the nucleic acids, polypeptides and
antibodies of this invention might be used to characterize the gene
expression of embryos. Ideally the stage-specific gene expression
of embryos of several different genotypes and at several different
stages of embryogenesis are characterized. For example, sets of
oligonucleotide primers designed using any one of SEQ ID NOS: 1-327
may be used in RT-PCR assays to characterize expression of a gene
product. In situ hybridization assays or antibody staining
protocols may also be used to characterize RNA and/or protein
expression and localization.
[0140] Embryos of the same genotype in which gene expression has
been characterized may also used be to generate plantlets that are
used in field testing. Once the embryos have developed into mature
trees, the various genotype trees can be evaluated for important
traits such as growth rates, herbicide tolerance, insect
resistance, pathogen resistance, climate and environmental
adaptability, wood quality, and wood fiber quality and content,
among others. Finally the phenotypic data collected from the field
testing can be correlated with gene expression during early
embryogenesis to further enhance the database of the present
invention. This will allow further identification of gene products
which whose expression is correlated, either positively or
negatively, with commercially valuable tree characteristics.
[0141] It will be clear to those skilled in the art that
identification of such gene products can have several uses.
Determining the correlation between a desirable phenotype and a
genotype would allow for the "pre-selection" of tree embryos for
field testing. It would also be useful in evaluating experimental
tissue culture conditions for somatic embryogenesis; in other
words, the expression level of a gene known to correlate with the
development of trees with desirable characteristics could serve as
the criterion on which culture media is evaluated, as opposed to
assessing the phenotype of fully matured trees. The ability to
evaluate culture conditions without having to develop fully mature
trees and do field testing would save a great deal of research time
and expense. And of course, the knowledge of the correlation
between gene expression and desirable tree phenotypes would serve
to identify target genes for genetic engineering.
[0142] Genetically Engineering Trees and Other Plants
[0143] There are several methods known in the art for the creation
of transgenic plants. These include, but are not limited to:
electroporation of plant protoplasts, liposome-mediated
transformation, polyethylene-glycol-mediated transformation,
microinjection of plant cells, and transformation using viruses.
Because the invention is especially concerned with the
transformation of woody species, the two prevalent methods for
transforming forest trees, namely Agrobacteriurm-mediated transfer
and direct gene transfer by particle bombardment, will be discussed
in more detail, though it is understood that the present invention
encompasses generation of transgenic plants via standard methods
commonly known in the art.
[0144] Agrobacterium Mediated Transfer
[0145] A. tumefaciens and A. rhizogenes are two soil microorganisms
that naturally infect a wide variety of plants including
dicotyledonous plants, gymnosperms and some monocotyledonous
plants. Infection by these organisms results in the growth of crown
gall tumors or in hairy root disease, respectively. Each of these
organisms carries a large plasmid, the tumor inducing (Ti) plasmid,
in the case of A. tumefaciens and the root-inducing (Ri) plasmid in
the case of A. rhizogenes. These plasmids have two critical
features, a set of virulence genes and a segment of DNA called
T-DNA that is delimited by conserved regions of approximately 25
base pairs known as the left and right borders. During infection,
the T-DNA is transferred to the plant cell where it is able to
stably integrate in single copy in the plant genome. Transfer of
T-DNA requires the function of the virulence genes.
[0146] In its natural state, T-DNA contains genes that mediate
progression of disease such as growth hormones or genes controlling
root morphogenesis. Using recombinant DNA technology, however,
T-DNA may be modified to contain an expression cassette encoding a
foreign gene of interest. There are several T-DNA vector systems
commonly in use for the transformation of plants. Several of these
vector systems are reviewed in Hansen et al., Current Topics in
Microbiology and Immunology 240: 21-57 (1999) which is hereby
incorporated by reference. T-DNA vectors must include the left and
right borders. In addition they must either be capable of
replication in Agrobacterium or be designed so as to recombine with
a plasmid that does so. The latter type of vector is known as a
co-integrate vector. For transformation to proceed, there must also
be a source of virulence (vir) genes. The vir genes may be on the
same plasmid with the T-DNA or more likely supplied by a helper
plasmid. For example, binary T-DNA vector systems are comprised of
two plasmids, one containing the vir genes and the other containing
T-DNA. Some plants known to be recalcitrant to
Agrobacterium-mediated transformation may be transformed if
additional copies of some or all virulence genes are provided.
Extra copies of VirG and VirE can be particularly useful.
[0147] Additionally, it is convenient to include in the T-DNA a
selectable marker that will allow identification and selection of
transformed plant cells. The selectable marker should be one that
works in both Agrobacterium and the target plant. For example, the
genes encoding chloramphenicol acetyltransferase and neomycin
phosphotransferase are suitable marker genes that confer resistance
to chloramphenicol and kanamycin, respectively. Additionally, a
selectable marker may be provided on a separate T-DNA from the
T-DNA encoding the gene of interest. Co-transformed T-DNAs can
integrate at separate sites in the plant genome. This can be useful
because it will later allow segregation of the marker gene in
progeny enabling the generation of transgenic trees expressing the
gene of interest but not the marker gene.
[0148] The gene of interest and the selectable marker genes must
also be under the control of promoters that function in the
transformed plant cell. Examples of suitable promoters include, but
are not limited to: the abscisic acid (ABA)-inducible promoter from
the early methionine (Em) gene from wheat (Marcotte et al., Plant
Cell 1:976-979 (1989); the cauliflower mosaic virus (CaMV) 35S
promoter (Odell et al., Nature 313:810-812 (1985); and the nopaline
synthase (nos) promoter (Sanders et al., Nucl. Acids Res.
15(4):1543-58 (1987). Tissue-specific plant promoters or plant
promoters responsive to chemical, hormone, heat or light treatments
may be used. Additionally, the gene of interest may be expressed
under the control of its endogenous promoter to ensure proper
regulation.
[0149] The process of transformation requires plant cells that are
competent and that are either embryogenic or organogenic. The plant
cells to be transformed are then co-cultivated with Agrobacterium
containing an engineered T-DNA vector system for 1-5 days.
Following the co-cultivation period, the cells are incubated with
the antibiotic against which the selectable marker confers
resistance, and transformed lines are selected for further
cultivation. The use of Agrobacterium mediated transfer in woody
trees is described in Loopstra et al., Plant Molecular Biology
15:1-9 (1990), Gallardo et al., Planta 210:19-26 (1999) and Wenck
et al., Plant Molecular Biology 39:407-419 (1999), each of which is
hereby incorporated by reference.
[0150] Direct Gene Transfer by Particle Bombardment
[0151] Direct gene transfer by particle bombardment provides
another method for transforming plant tissue. This method can be
especially useful when plant species are recalcitrant to
transformation by other means. In this technique a particle, or
microprojectile, coated with DNA is shot through the physical
barriers of the cell. Particle bombardment can be used to introduce
DNA into any target tissue that is penetrable by DNA coated
particles, but for stable transformation, it is imperative that
regenerable cells be used. Typically, the particles are made of
gold or tungsten. The particles are coated with DNA using either
CaCl.sup.2 or ethanol precipitation methods which are commonly
known in the art.
[0152] DNA coated particles are shot out of a particle gun. A
suitable particle gun can be purchased from Bio-Rad Laboratories
(Hercules, Calif.). Particle penetration is controlled by varying
parameters such as the intensity of the explosive burst, the size
of the particles, or the distance particles must travel to reach
the target tissue.
[0153] The DNA used for coating the particles should comprise an
expression cassette suitable for driving the expression of the gene
of interest. Minimally this will comprise a promoter operably
linked to the gene of interest. As with Agrobacterium mediated
transformation. Suitable promoters include, but are not limted to,
the the abscisic acid (ABA)-inducible Em promoter from wheat
(Marcotte et al., Plant Cell 1:976-979 (1989), the CaMV35S promoter
(Odell, et al., Nature 313:810-812 (1985), and the NOS:promoter
(Sanders et., Nucl. Acids Res. 15(4):1543-58 (1987).
[0154] Methods for performing direct gene transfer by particle
bombardment are disclosed in U.S. Pat. No. 5,990,387 to Tomes et
al. Additionally, Ellis et al. describe the successful use of
direct gene transfer to white spruce and larch trees in
Bio/Technology 11, 84-89 (1993).
[0155] Researchers skilled in the area of DNA or gene
transformation will recognize that additional procedures, or
combination of procedures, may be useful for the successful
tranformation of genetic stock.
[0156] Antisense Expression
[0157] The cDNAs of the invention may be expressed in such a way as
to produce either sense or antisense RNA. Antisense RNA is RNA that
has a sequence which is the reverse complement of the mRNA (sense
RNA) encoded by a gene. A vector that will drive the expression of
antisense RNA is one in which the cDNA is placed in "reverse
orientation" with respect to the promoter such that the non-coding
strand (rather than the coding strand) is transcribed. The
expression of antisense RNA can be used to down-modulate the
expression of the protein encoded by the mRNA to which the
antisense RNA is complementary. This phenomenon is also known as
"antisense suppression." It is believed that down-regulation of
protein expression following antisense RNA is caused by the binding
of the antisense RNA to the endogenous mRNA molecule to which it is
complementary, thereby, inhibiting or preventing translation of the
endogenous mRNA.
[0158] The antisense RNA expressed need not be the full-length cDNA
and need not be exactly homologous to the target mRNA. Generally,
however, where the introduced sequence is of shorter length, a
higher degree of homology to the endogenous mRNA will be needed for
effective antisense suppression. Preferably, the introduced
antisense sequence in the vector will be at least 30 nucleotides in
length, and improved antisense suppression will typically be
observed as the length of the antisense sequence increases. The
length of the antisense sequence in the vector may be greater than
100 nucleotides. Vectors producing antisense RNA's could be used to
make transgenic plants, as described above, in situations when
desirable tree characteristics are produced when the expression of
a particular gene is reduced or inhibited.
METHODS
[0159] Tissue Samples
[0160] The cDNAs of the current invention can be derived from any
sets of plant tissue. The cDNAs of SEQ ID NOS: 1-334, for example,
were originally derived from embryonic tissues of pine tree embryos
staged 1-9.9 as classified in Pullman and Webb TAPPI R&D
Division 1994 Biological Sciences Symposium, pages 31-34, which is
hereby incorporated by reference. LPS and LPZ clones are derived
from somatic and zygotic embryos, respectively. As noted, embryos
may be of either somatic or zygotic derivation, and the embryos may
be grown in either semi-solid or liquid tissue culture systems.
Applicable methods for growing embryos in semi-solid or liquid
tissue culture systems are disclosed in U.S. Pat. Nos.: 5,036,007;
5,236,841; 5,294,549; 5,413,930; 5,491,090; 5,506,136; 5,563,061;
5,677,185; 5,731,203; 5,731,204; and U.S. Patent Application
60/212,651 filed Jun. 19, 2000, which are hereby incorporated by
reference.
[0161] RNA Isolation
[0162] In one embodiment, RNA isolated from staged cell populations
provides the starting material for reverse transcription,
differential display, and cloning of amplified cDNA. Methods and
kits for isolating total RNA from cellular populations, or for
generating poly(A)+ RNA, are commonly known in the art. For
example, several procedures for isolating RNA are disclosed in
Chapter 4 of Current Protocols in Molecular Biology edited by F. A.
Ausubel et al., John Wiley and Sons, Inc. (1987) (incorporated
herein by reference). As an example, the TRI Reagent7 available
from Molecular Research Center, Inc. (Cincinnati, Ohio) is a
suitable reagent (used according to the manufacturer's
instructions) for isolation of RNA from plant tissues.
[0163] Differential Display
[0164] Differential display provides a method to identify
individual messenger RNAs that are differentially expressed among
two or more cell populations. In the practice of the present
invention, these cell populations may be provided by pine tree or
other plant embryos of different developmental stages. The
differential display procedure is taught in Liang et al., Science,
257:967-71 (1992) and in U.S. Pat. No. 5,262,311, which are hereby
incorporated by reference. Briefly, mRNA sequences are
PCR-amplified using two types of oligonucleotide primers known as
"anchor" and "arbitrary" primers. Anchor primers are designed to
recognize the polyadenylate tail of messenger RNAs. Arbitrary
primers are short and arbitrary in sequence ard anneal to
complementary sequences in various mRNAs. Products amplified with
these primers will vary in size and can be differentiated on an
agarose or sequencing gel based on their size. If different cell
populations are amplified with the same anchor and arbitrary
primers, one can compare the amplification products to identify
differentially expressed RNA sequences.
[0165] Cloning
[0166] PCR-amplified bands representing differentially expressed
RNA samples are excised from the gel, transferred to tubes and
reamplified using the same primer pairs and PCR conditions as used
in the differential display procedure. Methods for the cloning of
PCR products are commonly known in the art and there are several
commercially available reagents and kits for cloning PCR products.
For instance, the pCR-Scipt.TM. Cloning kit from Stratagene, La
Jolla, Calif.) is suitable for this purpose. Using this kit, E.
coli transformants containing plasmids with PCR fragment inserts
can rapidly be identified using blue/white color selection followed
by plasmid purification and restriction digests. The pCR-Script
vector contains T3 and T7 polymerase recognition sites allowing for
in vitro transcription of the inserted fragment.
[0167] Sequence Determination
[0168] Methods for sequencing DNA, including cloned PCR products,
are commonly known in the art. The selection of cloning vectors
having M13, T7 or T3 primer annealing sites flanking the
PCR-amplified insert can be used in sequencing reactions directly.
Most sequencing procedures in use today are modifications of
Sanger's dideoxy chain termination sequencing reaction as disclosed
in and Sanger et al., Proceedings of the National Academy of
Sciences, 74:5463-5467 (1977); which is hereby incorporated by
reference.
[0169] Homology Searching and Identification of Protein Coding
Sequences
[0170] As understood by one of ordinary skill in the art, the
sequence of a cloned cDNA insert obtained, may be compared against
public databases such as Genbank to discern any identity or
homology to known sequences. Programs, such BLAST, for performing
such a search are available on the National Center for
Biotechnology Information's web page located at
hftp://www.ncbi.nim.nih.qov. The results from Genbank search may
reveal the potential function of a polypeptide or RNA molecule
encoded by the cDNA. In addition to searching gene sequence
database, the use of commercially available analysis software is
well known in the art. For example, software packages such as the
Wisconsin Package.TM. (Genetic Computer Group, Madison, Wis.)
include programs such as FRAMES and CodonPreference that help to
identify protein coding sequences in a query nucleotide sequence.
FRAMES displays open reading frames for the six DNA translation
frames, allowing one to quickly assess the presence or absence of
stretches of open-reading frames that are likely to be protein
encoding regions. CodonPreference is a more sophisticated program
that identifies and displays possible protein coding regions based
on similarity of the codon usage in the sequence to a codon
frequency table (Gribskov et al., 1984).
EXAMPLE 1
Differential Gene Expression Analysis in Pine Tree Embryo
enesis
[0171] cDNA libraries were prepared from staged pine tree embryos,
as described above. The differential display technique was used to
identify 327 novel cDNAs that were preferentially-expressed during
early, middle, or late stages of pine tree embryogenesis, as set
forth below. Clone nomenclature is divided into subsets based on
tissue type; a clone is designated LPS to indicate somatic origins
and LPZ for zygotic origins.
[0172] Plant Materials
[0173] Somatic embryos were collected at different stages of
development. Cultures of somatic embryos of were initiated from
Loblolly pine immature zygotic embryos as described by Becwar et
al., Forestry Science 44:287-301 (1994) (incorporated by reference)
or with minor modifications in media mineral composition. Somatic
embryos were grown in cell suspension culture medium 16 (Pullman
and Webb, Tappi R&D Division 1994 Biological Sciences
Symposium) and a maturation medium similar to that of a standard
maturation media. Resulting somatic embryos were selected and
classified as stages 1-9 according to morphological development
following the teachings of Pullman and Webb, Tappi R&D Division
1994 Biological Sciences Symposium pp.31-34. Somatic embryos were
sorted into tubes containing the same stages and stored at
-70.degree. C.
[0174] RNA Isolation
[0175] Total RNA was isolated from all stages of somatic embryos of
loblolly pine and grouped into early, middle, and late phases of
development. The early phase is represented by a liquid suspension
culture containing embryos of stages 1 through stage 3. Middle
phase contains embryos of stages 4 through stage 6, while stages 7
through 9 formed the late phase. 60-100 mg aliquots of staged
frozen embryos were ground in 1.0 ml of TRI Reagent.RTM. Isolation
Reagent (Molecular Research Center, Inc.), a commercial product
that includes phenol and guideline thiocyanate in a monophase
solution and extracted according to the manufacturer's
instructions.
[0176] Reverse Transcription of mRNA (RT-PCR)
[0177] The total RNA was used as a template to synthesize single
stranded DNA mediated by MMLV reverse transcriptase (100 U/.mu.l).
The method involves the reverse transcription by PCR of the mRNA
with an oligo-dT primer (H-T.sub.11G: 5' B AAGCTTTTTTTTTTTG 3')
anchored to the beginning of the poly(A) tail, followed by a PCR
reaction in the presence of a second short (13-mer) primer which is
arbitrary in sequence [AP.sub.1 (5' B AAGCTTGATTGCC-3') or AP.sub.2
(5' B AAGCTTCGACTGT-3')]. Reverse transcription and Differential
Display were conducted using the GenHunter RNAimage Kit 1.
[0178] A 19 .mu.l reverse transcription reaction (10 .mu.l sterile
water, 2.0 .mu.l 5.times.RT buffer, 1.6 .mu.l dNTP (250 .mu.M), 2.0
.mu.l anchored primer (2.0 .mu.M), 2.0 .mu.l RNA template at 100
ng/.mu.l) was prepared for each embryo phase sample. The reaction
mixture was heated to 65.degree. C. for 5 minutes in a
thermocycler, cooled to 37.degree. C. and paused after 10 minutes
while 1.0 .mu.l MMLV was added. The program was allowed to resume
at 37.degree. C. for 50 minutes. The reaction was then heated to
75.degree. C. for 5 minutes, cooled to 4.degree. C. and stored at
-20.degree. C.
[0179] Incorporation of Radiolabeled Nucleotides by PCR
[0180] Differential Display PCR was performed in a 20 .mu.l
reaction containing 2 .mu.l of the reverse-transcribed cDNA
template; 10 .mu.l sterile water 2.0 .mu.l 10.times.PCR buffer, 1.6
.mu.l dNTP (25 .mu.M), 2.0 .mu.l anchored primer H-T 11G, (2.0
.mu.M), 2.0 .mu.l 13 mer arbitrary primer (AP.sub.1 or AP.sub.2
(2.0 .mu.M), 0.2 .mu.l Taq DNA polymerase, and 0.2 .mu.l
.alpha..sup.32P-dATP (2000 Ci/mmole). The cDNA was amplified by
PCR: 94.degree. C. for 3 minutes, 40 cycles of 94.degree. C. for 30
seconds, 40.degree. C. for 2 minutes, and 72.degree. C. for 30
seconds, followed by 72.degree. C. for 5 minutes. The reaction was
cooled to 4.degree. C. and stored at -20.degree. C.
[0181] Differential Display
[0182] The PCR products were separated on a Stratagene (La Jolla,
Calif.) pre-cast 6% polyacrylamide sequencing gel at 30 watts
constant power for approximately 2.5 to 3 hours. 3.5 .mu.l of
sample was mixed with 2.0 .mu.l, of loading dye and incubated at
80.degree. C. for 2 minutes immediately before loading onto the
gel. The gel was rinsed in water and dried. Dilute .sup.35P-dATP
with loading dye was spotted at the corners as alignment markers
and the gels were exposed to Kodak BioMaX.TM. autoradiography film.
An exemplary gel is shown in FIG. 1.
[0183] Bands that appeared to be possible markers for phase
specific gene expression were marked on the film and aligned over
the gel. The bands were excised by cutting through the film. The
gel pieces were scraped from the gel and transferred to tubes and
re-amplified using the same primer pairs and PCR conditions as used
for incorporation of radiolabeled nucleotides.
[0184] Cloning of DNA Fragments from Differential Display
[0185] The PCR products from the gel fragments were purified,
polished, ligated and cloned into XL 10-Gold Kan ultracompetent
cells by heat shock with the Stratagene pCR-Script Amp SK(+)
Supercompetent Cell Cloning Kit according to manufacturer's
instructions. The transformed cells were spread on LB agar plates
containing ampicillin, IPTG, and X-Gal each at 50 .mu.g/ml. The
plates were incubated overnight at 37.degree. C. Plasmids
containing PCR inserts were identified using blue-white colony
screening. The presence of inserts was confirmed by digesting the
clones with restriction endonucleases, Msc I and Nla ll, followed
by standard DNA gel electrophoresis. Transformants representing
early, middle, and late phase embryos were sequenced using standard
dideoxy protocols known in the art with the T3 primer.
[0186] Sequence Analysis
[0187] All sequences were analyzed using a program-database pair
search of the NCBI BLAST 2.0 server, blastn-nr, blastn-others ests,
and blastx-nr. In each case, the query sequence was filtered for
low complexity regions by default and entered in FASTA format.
Other formatting options were set by default; alignment
view-pairwise, descriptions-100, and alignments-50. Using these
parameter settings, significant similarity to known DNA, RNA, or
protein sequences was found for several of the nucleic acid
molecules of SEQ ID NOS: 1-334, for example, those described
herein. (Alignment data not shown).
EXAMPLE 2
Characterization of Full Length LP2-3 cDNA Sequence
[0188] SEQ ID NO: 327, designated LP2-3, was first identified
through differential display with T.sub.12MG and AP.sub.1 primers
(GeneHunter). The differential display band appeared to be present
only in liquid suspension cultures of Loblolly Pine somatic
embryos. The conditions for mRNA isolation, reverse-transcription,
differential display-PCR, and gel separation/visualization for
producing this band were all as described in Example 1. Likewise,
the band containing the original LP2-3 fragment was excised from
the differential display gel, amplified, and cloned into pCR-Script
AMP SK(+) according to standard protocols known in the art.
[0189] Northern Hybridizations Demonstrating Early-Specific
Expression
[0190] Northern analysis demonstrated that the LP2-3 differential
display clone hybridized to an approximately 1.2 Kb mRNA from
liquid suspension culture embryos but was undetectable in late
(6-9) stage embryo RNA. (FIG. 11) In general, LP2-3 is most highly
expressed in early stage embryos in liquid culture. LP2-3 mRNA is
found most abundantly in early stage somatic embryos, especially
for embryos grown in liquid multiplication medium. (FIG. 12)
Further, transcription decreases rapidly as embryos are transferred
to maturation medium (stage 3 and stage 4) and begin to mature.
LP2-3 transcripts are virtually undetectable at stage 6-9 somatic
embryos grown on maturation medium. (See FIG. 12) Additional
studies indicate that LP23 mRNA is expressed zygotically,
particularly in early stage zygotic embryos, but is undetectable in
mature vegetative tissues. (FIGS. 13 and 14) Specifically, the
signal intensity from liquid suspension somatic embryo RNA was
about 3 times greater than the signal from the analogous stage 1
zygotic embryo RNA. (FIGS. 13 and 14) LP2-3 transcripts were not
detectable in total RNA from needles, stems, or roots of one year
old seedlings, including those exposed to cold, ozone, wound
stresses, or the hormone jasmonic acid (not shown).
[0191] LP2-3 Differential Display and `Full-Length` cDNA
Sequences
[0192] A `full-length` cDNA was captured from SMART.TM. cDNA made
from somatic embryo liquid suspension by using a biotinylated LP2-3
differential display fragment as a capture probe. The "full-length"
cDNA was cloned and sequenced according to standard protocols known
in the art. This sequence was designated at LP2-3.sup.+.
[0193] GenBank blastx searches conducted with the above sequence
translated in all 6 reading frames indicated that LP2-3+likely
encodes a member of the major intrinsic protein family. This family
of proteins encodes membrane channels for the transport of water
and/or ions across cell membranes. They may play a significant role
in osmoregulation and may play a role in the cellular responses to
water and salt stresses. As is known in the art, the MIPs are
induced by dessication, flooding, and high levels of the plant
hormone ABA. In contrast, the LP2-3 sequence was not detected in
desiccated late-stage embryos which have high levels of ABA and,
thus, appears to be regulated by some embryo-specific signal.
EXAMPLE 3: Hypothesis Development for Improved Protocols
[0194] Currently the improvement of tissue culture practices arises
via hypothesis, evaluation and adoption. Hypotheses arise from
observation of size, shape, weight, etc. and physiological
measurement of ion or sugar content (FIG. 6, box 1). These
observations are limited in scope and this limits the improvements
that can be made to the tissue culture process. Gene expression is
closely linked to metabolic condition, thus the observation of
which genes are induced or repressed under a given growth
condition, naturally, on the tree, or in a culture vessel, provides
insight into the metabolic state of the embryo. This information
can be used to create new hypotheses that can be evaluated by
modifying tissue culture.
[0195] To this end, mRNA levels of two cDNAs (LPZ-202 and LPZ-216),
similar to "Late Embryogenesis Abundant" (LEA) proteins, identified
in other plants, were monitored. These genes are induced by the
plant hormone ABA. Two peaks of mRNA were observed in these clones
rather than the typical single peak in most plants. (See FIG. 4 for
clone LPZ-216; clone LPZ-202 is similar but data is not shown.) It
was subsequently confirmed that two peaks in ABA activity are
observed during development and that these correspond in timing to
the elevation in mRNA for LPZ-202 and LPZ-216. Thus mRNA abundance
profiles are providing insight into embryo physiology. (See FIG. 7)
The effect of giving two pulses of ABA to our somatic embryos is
assessed; a tissue culture modification that we might not have
considered as important had the gene expression data been
unavailable. Internal data shows fluctuations in the abundance of
mRNA for cDNAs listed in this collection (data not shown.)
[0196] Zygotic and Somatic Loblolly Pine Embryos
[0197] Loblolly pine cones were collected weekly from a breeding
orchard near Lake Charles, La., and shipped on ice for
experimentation. Embryos were excised and evaluated for
developmental stage (Pullman et al. 1994). Stage 9 embryos were
separated by the week they were collected-9.1 (week 1), 9.2 (week
2), etc. Staged zygotic embryos were sorted into vials partially
immersed in liquid nitrogen and stored at -70.degree. C. Somatic
embryos for loblolly pine were initiated as described by Becwar et
al. (1995) or with minor modifications. Somatic embryos were grown,
selected, and staged as described by Pullman et al. (1994) and
stored at -70.degree. C.
[0198] cDNA Probe Preparation and Hybridization
[0199] 30 ng of purified Lea protein cDNA fragments was labeled
with .sup.32P dCTP using the Ready-To-Go cDNA Random Labeling kit
(Pharmacia). The labeled cDNAs were purified using NICK Column
(Pharmacia) and heat denatured for hybridization. The RNA slot blot
was pre-hybridized in hybridization buffer (0.5 M sodium-phosphate,
pH 7.2, 5% SDS, and 10 mM EDTA) at 65.degree. C. for 2 hours in a
hybridization oven (Model 400, Robbins Scientific, Sunnyvale,
Calif.) and the hybridized in the same conditions with the cDNA
probes. After hybridization, the membranes were washed at
65.degree. C. in 0.2.times.SSC and 0.1% SDS. Each wash was 15 min.
The membranes were then exposed to Image Plate.
[0200] The probes can be stripped from the RNA slot blot by pouring
boiling 0.5% SDS onto the membrane twice and incubating without
heating for 30 min. The stripped blot was then exposed to Image
Plate for overnight to check the completeness of the de-probing
before next round of hybridization.
[0201] To ensure the equal loading of the each RNA sample, the same
membranes were stripped and hybridized with a .sup.32P-dCTP labeled
26S ribosomal rDNA fragment. These results were used as controls to
normalize the Lea protein gene expression levels.
[0202] As a means of evaluating the usefulness of these arrays, we
followed the expression of three cDNAs that have strong sequence
similarity to late embryo-abundant proteins, (Lea) proteins from
cotton (Baker et al 1988). Lea proteins and mRNAs appear in embryos
at a stage when ABA is high and the genes can be induced in
vegetative tissue by application of ABA. The transcript level of
Lea genes LPZ-202 and LPZ-216 showed two peaks, rising from stage 5
and returning to a base line about stage 9.2 then rising again
around stage 9.5. (See FIG. 4 for clone LPZ-216).
[0203] To confirm the fluctuation in lea transcript levels by
Northern analysis. RNA was extracted from zygotic embryos at
different stages of development A in `dehydrin` cDNA from the North
Carolina State University cDNA collection
(hftp://www.cbc.med.umn.edu/ResearchProiects/Pine/DOE.pin-
e/index.html) was used as probe for some experiments. Dehydrins are
a class of lea protein, originally identified as water deficit
inducible proteins. Since the expression of this class of protein
is well characterized, in contrast to our lea genes, the dehydrin
expression profile could act as a reference point. After probing
with dehydrin, blots were stripped and probed with a 26S rDNA probe
from Arabidopsis to check the loading of the original gel. The
normalized expression pattern of dehydrin in the zygotic
embryogenesis is illustrated in the top panel of FIG. 4. The
expression of the dehydrin gene was induced at stage 5 and reached
a peak at stage 6. It declined at stage 7-8, just prior to the
onset of the desiccation. Then the mRNAs level remained low from
stage 9.1 through 9.5. The dehydrin mRNA levels rose again late in
development, from stage 9.6 on, apparently dropping in very late
development. A similar pattern of expression was observed in a
parallel experiment when our lea-like clone, LPZ-216, was used as a
probe.
[0204] This pattern reveals two significant peaks at the early
development of the embryos and high expression levels for the stage
9.6 and beyond. The expression pattern of these two lea genes in
loblolly pine embryos is consistent with the changes in ABA
concentration observed in pine during embryogenesis. (See FIG.
5)
EXAMPLE 4
Evaluation of Metabolic State of Somatic
[0205] Embryos as Compared to Zygotic Embryos for Fitness
Determination
[0206] The model and goal for somatic embryogenesis is to produce
an embryo that in vigor, germinatability, etc., resembles a zygotic
embryo. Standard measurements reveal relatively little about the
embryos; thus the metabolic state of somatic and zygotic embryos is
unknown. The metabolic state of zygotic (natural) embryos can be
evaluated by DNA arrays containing the cDNA clones described in
this application. A database of mRNA levels for the genes
represented on the DNA arrays can then be established. Embryos
growing under a new tissue culture protocol (FIG. 6, box #2) can be
evaluated by DNA array southerns (FIG. 6, box #3). The array
elucidates patterns of gene activity and reveals whether those
patterns are similar to the natural state (FIG. 6, box #4). The
germination, or further development can be checked (FIG. 6, box #5)
to confirm the conclusion. Where a link between specific gene
activity and embryo performance has been demonstrated, protocols
can be modified with efficiency as seen in FIG. 6, box 6.
[0207] To illustrate this process, elevation of plant hormone ABA
in maturation medium was evaluated as a protocol modification, as
described below. This modification proved beneficial, elevating the
number and quality of the embryos produced. The mRNA abundance for
cDNAs was assessed by DNA array using RNA isolated from control and
elevated ABA conditions; several differences were observed in the
mRNA levels of specific genes. Further, abundance of mRNA in the
elevated ABA condition, more closely resembled the mRNA abundance
observed for the these same genes in zygotic embryos. Thus a
protocol which produces higher quality embryos produces, in these
embryos, a mRNA profile that more closely resembles that observed
in natural embryos.
[0208] Zygotic and Somatic Loblolly Pine Embryos
[0209] Loblolly pine cones were collected weekly from a breeding
orchard near Lake Charles, La., and shipped on ice for
experimentation. Embryos were excised and evaluated for
developmental stage (Pullman et al. 1994). Stage 9 embryos were
separated by the week they were collected-9.1 (week 1), 9.2 (week
2), etc. Staged zygotic embryos were sorted into vials partially
immersed in liquid nitrogen and stored at -70.degree. C. Somatic
embryos for loblolly pine were initiated as described by Becwar et
al. (1995) or with minor modifications. Somatic embryos were grown,
selected, and staged as described by Pullman et al. (1994) and
stored at -70.degree. C.
[0210] Mass Isolation of Genes Differentially Expressed in Loblolly
Pine Zygotic Embryos
[0211] The following RNA differential display method is sensitive
enough to produce banding patterns from one mid- to late-stage
embryo or 10-20 early stage embryos. This technique, which extracts
mRNA directly from tissue using oligo(dt) beads, avoids losses
inherent in conventional RNA extraction methods, is fast, reliable,
and inexpensive. Differences in gene expression during development,
as well as between somatic and zygotic embryos, can be easily
detected.
[0212] To achieve these results, 50-100 .mu.l lysis buffer
containing 100 mM Tris-HCl, pH 8.0, 500 mM LiCl, 10 mM EDTA, 1% SDS
and 5 mM DTT was added to 10-100 mg of staged embryos in a 1.5 ml
tube. The mixture was ground thoroughly with an electric drill
containing a plastic pestle bit (VWR, Cat# KT95050-99) that had
been sterilized by autoclaving. An additional 50-100 .mu.l lysis
buffer was added and ground briefly. The grinder and vortex was
washed with 100 .mu.l lysis buffer. If multiple samples were
processed, each is stored on ice until ready for the next step. The
grinding tip was washed with sterile water and dried for the next
sample.
[0213] After all the samples were ground, they were spun at
4.degree. C. for 15 minutes in a bench top centrifuge at 14,000
rpm. 8 .mu.l oligo(dT) coated Dynal beads (mRNA DIRECT Kit, Dynal,
N.Y.) was placed in a 1.5 ml tube. The Dynal beads were washed
twice with a 100 .mu.l of the above mentioned lysis buffer and
suspended in an equal volume of the lysis buffer used in tissue
grinding. If more than one sample is handled, the beads for all the
samples can be washed together and dispensed in several 1.5-ml
tubes. The cleared embryo lysate (after centrifugation) was added
to the beads and mixed well.
[0214] The mixture was then incubated on ice for 5 min., placed on
a magnetic stand (Promega) for 5 min., and partially dried by
careful removal of the liquid. To this, 100 .mu.l of washing buffer
with LiDS containing 100 mM Tris-HCl, pH 8.0, 0.15 mM LiCl, 1.0 mM
EDTA, and 0.1% SDS was added, (mRNA DIRECT kit.) The mix was
transferred to a 200 .mu.l PCR tube. The beads were washed once
with 100 .mu.l washing buffer with LIDS and once with 50 .mu.l
washing buffer containing 100 mM Tris-HCl, pH 8.0, 0.15 mM LiCl,
and 1.0 mM EDTA. (mRNA DIRECT kit.) The beads were then washed
quickly with 20 .mu.l 1.times.RT Buffer (25 mM Tris-HCl, pH 8.3,
37.6 mM KCl, 2.5 mM MgCl2, and 5 mM DTT) and 20 .mu.l RT Mix
containing 1.times.RT Buffer and 20 .mu.M dNTP was added. The tube
was heated at 65.degree. C. for 5 min. and cooled to 37.degree. C.
1 .mu.l MMLV reverse transcriptase (Promega) was added and the
mixture was incubated at 37.degree. C. for 1 h. with occasional
shaking. Next, 20 .mu.l of water was added to the RT reaction,
mixed and a 1.0 .mu.l to 20 .mu.l aliquot of the PCR mix containing
1.times.Perkin-Elmer PCR buffer, 2.0 .mu.M dNTP, 1.0 .mu.M T12VN,
0.2 .mu.M arbitrary 10-mer, 1 unit AmpliTaq (Perkin-Elmer), 50
.mu.Ci .alpha..sup.35S-dATP (Amersham) was taken. PCR using
temperature settings of 94.degree. C. 30", 40.degree. C. 1',
72.degree. C. 2', 40 cycles, and 72.degree. C. 10' extension was
performed with the Perkin Elmer 9600 Thermal Cycler. All PCR
product was run on appropriate gels for band visualization.
[0215] cDNA cloning of Differential Display Bands
[0216] All dried gels were marked with radioactive ink prior to
film exposure for proper alignment between the X-ray film and the
dried gel plate. Appropriate bands were marked by puncturing. A
scalpel blade was used to score the gel around each band to be
excised. The excised gel pieces were placed into a PCR tube
containing 2 .mu.l water. PCR was performed using a 50 .mu.l PCR
mix (same as for differential display with the following
modifications: the primer concentration was 1 .mu.M, and the dNTP
concentration was 200 .mu.M; no .alpha..sup.35S-dATP is added.) The
cycle settings were the same as above.
[0217] A portion of the PCR products was run on a gel to determine
amount and size of PCR products; DNA that did not correspond to the
size of the original differential display band was discarded. The
remaining PCR fractions were purified using CHROMA SPIN-100 columns
(Clontech, Palo Alto, Calif.) according to the manufacturer's
instructions. The purified PCR fragments were cloned into the
pCR2.1 TA cloning vector (Invitrogen) according to Invitrogen
cloning protocols supplied with the vector. The only variation from
the standard protocol was an increase in the molar concentration of
PCR product to vector (over 100-fold); multiple insertions were not
found to be a problem. All ligations were performed at 16.degree.
C. overnight, transformed into E. coli strain DH5.alpha., and
plated onto LB with X-gal/IPTG.
[0218] Five colonies were chosen for PCR verification; PCR products
of expected size were selected. About 10 .mu.l of the 30 .mu.l PCR
reaction was simultaneously digested with Nla III and Mse I
overnight at 37.degree. C. (a 5 h digestion was used as well.) cDNA
clones were selected according to the colony PCR and the
restriction enzyme digestion pattern.
[0219] The differential display protocol for finely staged zygotic
embryos of loblolly pine as described above, has produced more than
600 differential display patterns and more than 60,000 bands.
Within that set of bands, we have identified bands that increased
and/or decreased during embryo development. From those bands cDNA
clones of this invention were isolated and sequenced.
[0220] Detection of Gene Expression by Micro-Array Assay
[0221] In order to verify expression patterns of the cloned DNA in
loblolly pine embryos a micro-array assay was developed. The cloned
cDNAs were amplified by PCR and adjusted to equal concentrations
(0.1 .mu.g/.mu.l). The cDNAs were then dispensed in the wells of a
384-well plate, denatured in 0.3 M NaOH at 65.degree. C. for 30
min. and neutralized with 2 volumes of 20.times.SSPE mixed with
0.00125% bromophenol blue and 0.0125% xylene cyanol FF (5% gel
loading dye). The denatured DNAs were then blotted on to Hybond
N+membranes (Amersham) as arrays using a VP 386 pin blotter
(V&P Scientific, Inc., San Diego, Calif.). Each DNA was
dot-blotted four times as a quartet on the membrane. An example of
quartet spotting is seen in FIG. 7. Each dot is about 1.2 mm in
diameter and contains about 3 ng of DNA. DNA was then cross-linked
to the membrane at 120,000 mJ/cm2 in a CL-1000 UV-linker.
(Strategene, Inc., Upland, Calif.) The dot image of each membrane
was scanned, numbered and saved in computer for later use in data
digitizing.
[0222] The cDNA array membranes were pre-hybridized in
hybridization buffer (0.5 M Na-phosphate, pH 7.2, 5% SDS, and 10 mM
EDTA) at 65.degree. C. for 30' in a hybridization oven (Model 400,
Robbins Scientific, Sunnyvale, Calif.) and then hybridized under
the same conditions with total cDNA probes made from mRNA. The
membranes were washed twice at room temperature in 2.times.SSPE and
0.1% SDS, twice in 0.5.times.SSPE and 0.1% SDS, and twice in
0.1.times. hybridization buffer. Each wash was roughly 20 min. Each
membrane was then exposed to Kodak Biomax MR films.
[0223] The total cDNA probes referred to above were made by
initially creating the first strand cDNA. This was accomplished by
mixing loblolly pine embryos (0.05-0.1 gm fresh weight) with 100
.mu.l lysis buffer (containing 100 mM Tris-HCl, pH 8.0, 500 mM
LiCl, 10 mM EDTA, 1% SDS and 5 mM DTT) in a 1.5 ml Eppendorf tube.
The mix was then ground with an electric drill as described above.
Another 100 .mu.l lysis buffer was added and the lysate was ground
again briefly. The drill pestle was washed with 100 .mu.l lysis
buffer that was pooled with the lysate. After centrifugation at 14K
at 4.degree. C. for 15 min. in a Beckman bench top centrifuge, the
clear embryo lysate was mixed with 10 .mu.l Dynal beads washed
twice with lysis buffer. The suspension was incubated on ice for 5
min., with occasional mixing to allow binding of Poly (A) RNA to
the oligo (dT) on the beads, and then left on a magnetic stand at
room temperature for another 5 min. The liquid was removed and the
beads were moved to a 0.2 ml PCR tube by suspending in 100 .mu.l
lysis buffer.
[0224] The beads were washed twice with 100 .mu.l of washing buffer
with LiDS and once with 50 .mu.l of washing buffer. The mRNA was
eluted from the beads in 6 .mu.l water at 65.degree. C. for 2'. One
.mu.l T21VN primer (10 .mu.M) and 1 .mu.l SCSP oligo (cap switch
primer, 5'-ctcttaattaagtacgcggg-3', 10 .mu.M) were added to the
mRNA eluate. The mixture was incubated at 70.degree. C. for 2' and
cooled on ice. Three .mu.l 5.times.First Strand Buffer, 1.5 .mu.l
DTT (20 mM), 1.5 .mu.l dNTP (10 mM each) and 1 .mu.l MMLV
Superscript II (Gibco BRL) were added to the mRNA-primer mixture
followed by incubation at 42.degree. C. for 1 h to synthesis first
strand cDNAs. The cDNA was heated to 72.degree. C. for 1 min. to
degrade RNA and then diluted to 100 .mu.l with water. The lysis
buffer, washing buffer and Dynal beads are components of the mRNA
DIRECT kit (Dynal, N.Y.). The first strand buffer (5.times.), 20 mM
DTT and 10 mM dNTP are components of the SMART PCR cDNA synthesis
kit (Clontech, Palo Alto, Calif.).
[0225] The first strand cDNAs synthesized as described above
contains a T21VN sequence at their 5' ends and the SCSP sequence
(see "SMARTTM cDNA, Clontech, Palo Alto, Calif.) at their 3'
terminals. Total cDNA probes were made by PCR amplifying the first
strand cDNAs using SMART cDNA PCR (Clontech, Palo Alto, Calif.) in
the presence of labeling agent. Five 5 .mu.l first strand cDNA
solution was mixed with 5 .mu.l 10.times.KlenTaq PCR buffer
(Clonetech), 5 .mu.l dATP+dGTP+dUTP (5 .mu.M each), 1 .mu.l T21VN
primer, 1 .mu.l SCSP oligo, 1 .mu.l KlenTaq Mix, 5 .mu.l
.sup.32P-dCTP (10 mCi/ml, Amersham) and 27 .mu.l water. The PCR was
performed using the setting of 94.degree. C. 2', 15 cycles of
95.degree. C. 15", 52.degree. C. 30", 68.degree. C. 6'. The PCR
products were purified using NICK column (Pharmacia) according to
the manufacture's instructions.
[0226] Currently, high-density array Southerns for both somatic and
zygotic embryos at all the developmental stages have been
performed. The dot array Southern data indicate that gene
expression of late stage somatic embryos resembles middle stage
zygotic embryos; many transcripts present during late zygotic
embryogenesis (ZE) are absent in somatic embryos and late stage
somatic embryo gene expression patterns resemble the patterns of
middle stage zygotic embryos.
[0227] Cairney et al. (In Vitro Cell. & Devel. Biol.-Plant.
36:155-162 (2000); Appl. Biochem. Biotech. 77-79:5-17 (1999)) have
discussed how this gene expression information may be used to
improve the process of somatic embryogenesis; the rare incorporated
in their entirety. As shown in FIG. 2, the high-density array
Southerns allows rapid evaluation of embryos subjected to protocol
changes. Following the expression of a known gene permits
inferences about metabolism and is very valuable in developing
media-improvement hypotheses. Further, detailed gene expression
studies may help by providing an understanding of the timing and
location of gene expression (e.g., in situ hybridization). The
isolation of key genes also provides the ability to monitor the
expression of these genes as stage specific markers and allows
protocol variations to be quickly evaluated.
EXAMPLE 5
Identification of Markers for Superior Performance in Tissue
Culture"
[0228] The evaluation of tissue culture modifications for pine
somatic embryogenesis, depicted in FIG. 8, is typically a lengthy
process. However, where molecular tools are available, potentially
improved media or genotypes can be discerned more rapidly, thereby
avoiding the months of costly evaluation. (See FIG. 8) Table 5
illustrates this proposition.
[0229] Table 4 describes several publicly available clones. Lec.
Fie, and Pkl, used to provide a representative model for this
example. Any clone within Table 1, SEQ ID NOS: 1-327, can be
substituted for those in Table 4 to assay increased performance in
tissue culture. Any promoter within Table 1, SEQ ID NOS: 328-334,
can be incorporated with those in Table 4 or SEQ ID NOS: 1-327 to
assay increased performance in tissue culture. In this scenario,
Table 5, a representation of the information contained in FIG. 9,
shows performance of selected genotypes (260, 480, 499, and 500) in
various media (1133 or 16) determined by the total number of
embryos produced per medium as described by Pullman and Webb
(1994), incorporated herein. Embryo maturation was determined by
the presence of recognized morphology according to methods
previously mentioned above. (Pullman and Webb, (1994)) Genotypes
that produced high, medium, and low numbers of embryos were
selected for RNA extraction. Gene expression assays, such as DNA
arrays, Northern blots, slot blots, etc., were used in attempt to
correlate embryo performance with mRNA abundance for selected
genes. In the example shown in FIG. 9 and Table 5, expression of
loblolly pine genes, designated as Lec, Fie, and Pkl, obtained from
the Pine Gene Discovery Project, was evaluated. The preliminary
correlation appears to be that the high levels of the Lec gene's
mRNA correlates with greater number of pine embryos. (See table 5.)
These experiments can be further expanded to incorporate additional
or alternative genotypes with the prospect of identifying a large
collection of gene indicators of good or poor performance in tissue
culture based on high or low mRNA levels. It is clear from the
above that this approach, using the sequences disclosed in this
application, can evaluate a genotype entering tissue culture,
saving both time and expense.
[0230] Somatic Embryos
[0231] Immature zygotic seeds were collected from loblolly pine
genotype 260 (mother tree BC-3, Boise Cascade). Somatic embryos
were initiated as described by Becwar et al. (1990) or with
modifications in media mineral composition. The early stage somatic
embryos were grown in cell suspension culture medium 16 and
sub-cultured every week (Pullman and Webb, 1994). The embryos
collected from the suspension, which include stage 1 and stage 2
somatic embryos, are referred to as stage S embryos. At the end of
the subculture week, the somatic embryos in the suspension were
settled in a cylinder and transferred to maturation medium 240
(Pullman and Webb, 1994). Resulting somatic embryos were selected,
staged, sorted into vials containing the same stage, and stored at
-70.degree. C. until analyses were performed.
[0232] Probes
[0233] For the following example analysis RNA was isolated from
embryos at different stages in development, early stage somatic
embryos and late-stage somatic embryos. The cDNA probes used in
this example are not contained in the SEQ ID NOS: 1-327, but
rather, are generic, publicly available pine sequences obtained
from the Pine Gene Discovery project located at
(http://www.cbc.med.umn.edu/ResearchProiects/Pine/DOE.pine/ind-
ex.html). These clones are homologs to the well-studied Arabidopsis
genes that have been shown to have significant influence on embryo
development in this plant. The pine clone names (first column) and
corresponding references for the Arabidopsis homologs are shown in
Table 4. The three clones listed, Lec, Lie, and Pkl, are for
representative purposes within this example and it will be clear to
one skilled in the art that any of the SEQ ID NOS: 1-327 could be
substituted for those here as all will help identify conditions for
improved performance in culture.
[0234] Probes were made by preparation of DNA using Wizard
Minipreps (Promega, Madison, Wis.) and cDNA inserts isolated by
restriction enzyme digestion. For the cDNA probes, 50 ng of the
isolated cDNA insert DNA was used to make .sup.32P-labeled probes
with Ready-To-Go DNA labeling beads (Amersham Pharmacia Biotech)
according to manufacturer's instructions. Blots were prehybridized
(7% SDS, 1% BSA, 0.25 M NaPO.sub.4 (pH 7.2), 1.0 mM EDTA) for 3
hours at 65.degree. C. and hybridized in fresh buffer at 65.degree.
C. for 12 to 18 hours (4). Each blot was washed 6 times with the
following conditions: 1) RT, 2.times.SSC, 0.1% SDS, 15 min; 2) RT,
2.times.SSC, 0.1% SDS, 30 min; 3) 42.degree. C., 0.2.times.SSC,
0.1% SDS, 15 min; 4) 42.degree. C., 0.2.times.SSC, 0.1% SDS, 30
min; 5) 60.degree. C., 0.2.times.SSC, 0.1% SDS, 30 min; 6)
60.degree. C., 0.2.times.SSC, 0.1% SDS, 30 min. Blots were exposed
to a phosphorimaging plate for 10 minutes. Screens were read with a
BAS1800 (software v1.0) and images were manipulated with ImageGauge
(v2.54) (Fuji Photo Film Co., Ltd., Kanagawa, Japan).
[0235] The hypothesis tested within this example is that genotypes
that produce large numbers of embryos have high Lec expression and
low Pkl expression, poor genotypes have the opposite pattern, and
that Lec and Pkl expression act as indicators of embryogenic
potential. FIG. 9 shows that Lec is not expressed in late stages of
embryogenesis in somatic embryos. The Lec gene is expressed
throughout embryogenesis in Arabidopsis. The blot reveals that the
Lec gene is a useful early expression marker for embryogenesis. One
interpretation of these results is that the somatic embryos do not
express Lec in the manner that Lec is expressed in zygotic embryos,
i.e. the use of Lec expression has highlighted a defect in gene
expression in somatic embryos. This defect could be used to
identify desirable genotypes, i.e. those likely to progress through
development and produce a large number of healthy plantlets
compared to undesirable genotypes that will cease development
prematurely or produce low numbers of plantlets. This is an example
of the principle described pictorially in FIG. 8.
[0236] The results described in the previous section of Example 5
reveal ways in which gene expression analyses can be used to
improve somatic embryogenesis based on several genes. However, this
principle applies as well when the assay is expanded to determine
the expression of hundreds or thousands of genes simultaneously
(e.g. by DNA arrays). We can create hypotheses which state that
expression of a single specific gene can be used to determine the
potential of a culture, or hypotheses that state that the
expression of a group of genes (e.g., hypothetical genes A, B, C,
D, E, F) acts as an indicator of high embryogenic potential. For
example, all these genes may be expressed at a high level in cell
lines that produce large numbers of embryos, thus we would select
cell lines which exhibited this characteristic. Alternatively
specific levels of expression for genes A, B, C, D, E and F may be
required and a combination of high and low expression of particular
genes will identify desirable cultures. Alternatively, experience
will determine that certain exceptions can be tolerated.
[0237] While the previous paragraphs discuss numbers of embryos
produced, the principle applies to ANY desired characteristic: by
establishing a correlation of gene expression with e.g.,
germination potential, embryo size, growth of plantlets in their
first year, disease resistance of mature plants, environmental
hardiness or wood quality. Any trait where could be evaluated by
these gene expression assays and correlations with gene expression
established, resulting in a molecular tool which could be used to
predict desirable characteristics. Explicitly, we could use these
gene expression tools to select cell lines which will produce high
quality plantlets months before they grow into plantlets, or cell
lines or juvenile plantlets which will produce hardy trees with
desirable wood quality, years before these traits are
expressed.
1 TABLE I Embryo cDNA Phase Clone Nucleotide Sequence SEQ ID NO:1
Late LPS-001
GGTACTCCACCGTAATAACCCTTGGGAAATAGCCTATGATCCAGGGGAGGCAACC
ACCTATATCATTGACAACAGCGAAAAATGTGGCGCAAGAAGTTTCACATACAATTCA
TGGTTACAAAGATCACATACCAGGTGTTGGAGCAGATTCGATAGATATTGAAGATAT
GAAGCCAAGGAGTGGAGCAGTTATTGAAAAGGGCACAAAAAAATTTGCCATTTACA
AAGATGAAAATGGGCTGATTCACAAATACTCGGCAATATGCCCACACATGAACTGT
ATTGTGAAATGGAATCCTATAGACTCAACTTTCGATTGCCCCTGCCATGGTTCAATG
TTTGATAATCTGGGTCGATGCATCAATGGACCTGCCAAGGCGGACCTATTTCCCGA
AGATTAACGATAGTTGTTTGTACATGTAATTATCTTGATATTGTATATATATGTAT- TTA
AATTATACAGTACAATAAATCCATGTTGCAGGCTATTTCTGCTTGATAATT- TAGCTC
CAGATTATACATAACCAGTTATTGGCTGTTTTCCCCTGGCAAAAAAAA- AAAA SEQ ID NO:2
Late LPS-003 003GGTACTCCACAGAAAGAAATGATT-
TGACAGAAAAAGAGAGCTGTAGGATTGGG AAACCCTGCAGTGGATATATACAAT-
GTATATGTACTCTGTCTGTTTTTCTGTTATTTG
ACGGAAATAAAAACGCCATAGCGACGGATGACTGTAAATCCTTAGGGACGGATGAC
TGTAAATCCTTAGGTTGGAAGATTACAAACGACATATGGGTCTTTCAATTTTCAGAT
TTCTGTAAGACTTACATTTCAAAGACTGTTTGGATGGGCAAAAAAAAAAAA SEQ ID NO:3
Middle LPS-004 GGTACTCCACCAGAATGCCGCAGTTTAGTTCTCTAAAGCAAGCAGT-
AAATTAATTTT GTCAAAATCTAAAGAGTGTATAGTATCAGTGGGTTTGTATTTC-
CTAGTTTGCCTACA ATAACGATGGGGATTCACCAGTTTTTGTAGAATTTGCAAT-
CATCGGATGACAATTTC AAAGTTTTCTCTAAGTCACCCGCATTGATATCGAGAA-
GCCTTCCATTTTCAATTATTT AATATCAGAAAATCTTTTCAGTTGGCAAAAAAA- AAAAA SEQ
ID NO:4 Middle LPS-006 AGCCCAGCTGCGAAGGGGATGTGCT-
GCAAGCGATAAGTGGTAACGCCAGGTTTCC AGTCAGACGTGTAAACGACGCCAG-
TGATGTATACGAATCACTATAGGCGATGGCCT
TCTAGATGCATGCTCGAGCGCCGCAGTGTGATGAATTGCAGAATCGGCTGGTACT
CACGGGCTAGAGAAAGGCACAAGCACTTTTTGTCATTTTAGGATCAGAGGCATTCA
GGTATAGGAAGGGTGGCTCAGATAGGCAGATGGATCGGCATTTTGCCCAGTCATG
AAACATTTTATGCATGTTATTGCCTCCCAAGGACGAAATCAGTTCTTTGTGCCTTCT
GGTGATATCACTTCAAACAAAAGGCAACAGTTCTGTGATTTCATATGGTTTGTCACT
GAATATTTTGTTGCAGATGTTCTCTACTATTTTTTATCTGCTTTCAAGTGATTATTTG
TTGATTCCCCATGGATAGTTATGCTAATCAGTTGCATTTCTCTTGTACCAGTCAACA
AACAAAAATGCTTGTAGGAATCCATTACTATTTATTTTCAGACAGGTAAACGTGTA- G
CTAATTGTTCTGGCAAAAAAAAAAAA SEQ ID NO:5 Middle LPS-007
TCCAAAATACAAAGGCTTTATTTGCATCATGATATAATACAAAGTAAGAAATT- TACCC
AACTGTTTAACCTAATAATAATACAAAGGAAGCATTTTACCCAACTCTT- TAACGTAAT
AATACCAAAGAGTGGAATGCTTTATTGACCAGCAAGACCTTGAAA- TTTTTATAACCA
ATGCCCATCAACAGAGCCTTTCTTAAAAAACGCAAAGCCCAG- CTCTGTCACCTTATT
AGTTAGTATAAACTGACATTCTTCCAAGCTTGTGTGCGC- AGAAACAATAAAGAACT
CACCTTGGTTTAAAGAACGTGCCATGAAGAAAACGTC- CCAAGAAAAATGAAATGGC
TCCTTCGACCATTCAGTCCTCCCTAGAAAAATCAA- AAGACTCCTTCGACCATTAGGT
CCTCCAATTGGGCATCTAACTACAAGCGGTC SEQ ID NO:6 Middle LPS-008
GGTACTCCACGGGCTAGAGAAAAGGCACAA- GCACTTCTTCGTCATTTTAGGGATCA
GAGGCATTCAGGTATAGGAAGGGGTGGC- TCAGATAGGCAGATGGATCGGCATTTT
GCCCAGTCATGAAACATTTTATGCATG- TTATTGCCTCCCAAGGACGAAATCAGTTCT
TTGTGCCTTCTGGTGATATCACTT- CAAACAAAGGCAACAGTTCTGTGATTTCATAT
GGTTTGTCACTGAATATTTTGTTGCAGATGTTCTCTACTATTTTTTATCTGCTTTCAA
GTGATTATTTGTTGATTCCCCATGGATAGTTATGCTAATCAGTTGCATTTCTCTTGTA
CCAGTCAACAAACAAAAATGCTTGTAGGAATCCATTACTATTTATTTTCAGACAGGT
AAACGTGTAGCTAATTGTCTGGCAAAAAAAAAAAA SEQ ID NO:7 Middle LPS-010
ACGACGTGTAAACGACGGCCAGTGATTGTATACGACTCACTATAGGGCGATTG- GC
CTTCTAGATGCATGCTCGAGCGGCCGCAGGTGATGGATATCTGCAGAATTCG- CTT
GGTACTCCACGGCTAGAGAAAAGGCACAAGCACTTCTTCGTCATTTTAGGA- TCAGA
GGCATTCAGGTATAGGAAGGGTGGTCAGATAGGCAGATGGATCGGCATT- TTGCCC
AGTCATGAAACATTTTATGCATGTTATTGCCTCCCAAGGACGAAATCA- GTTCTTTGT
GCCTTCTGGTGATATCACTTCAAACAAAAGGCAACAGTTCTGTGA- TTTCATATGGTT
TGTCACTGAATATTTTGTTGCAGATGTTCTCTACTATTTTTT- ATCTGCTTTCAAGTGA
TTATTTGTTGATTCCCCATGGATAGTTATGCTAATCAG- TTGCATTTCTCTTGTACCAG
TCAACAAACAAAAATGCTTGTAGGAATCCATTAC- TATTTATTTTCAGACAGGTAAAC
GTGTAGCTAATTGTTCTGGCAAAAAAAAAAA SEQ ID NO:8 Middle LPS-011
GGTACTCCACGAAGCAAAAAGAGTCAGGGG- AATGAAGATGGGGGGCTCCGACAAG
AAGCGGATCAGAGAAGAGCAGGAAATGAG- TCCACCTGAGGAATCCTGGAGACAGA
AACAGGGGCGTTTTAATGGAGTTTGAGG- CAGGGATGGCCTATGATAAACCTGAAAAT
GCCGGTGCAGGTAATGAGAATTTGC- CAGAGTTTTGCTCTCTTTCAAATGAGTACTC
GATGTTATTGAAAGATCCATGGA- GTTGGGAGGATAGCACTGGTTTCGGAATCCGAA
GCTTAGCTGCTGTCAGGAAGCAGTCTTGTATATTGGACTATCTCCATGATTCTGCT
GTAGATAATCGCTGTGAAAAGGATTTTGCCGAGCAGCACAAGGTACAGGAAGAGG
AGGATTGTTTGAGAAGGTCTCTTTTTGAAGCCACAGATGATCAGCTCTGGAGGCTT
CAGAGTCTTTGCAGGATACAGAAGGTCTGTTTCCTCTGGATTCCGTGGGTAGCCAT
GATTGCACGACCTTGTTGCAGGATGAGAGCATTGTTCAGGGCGCTGCTCTTACTT
CAGAATTTGGGAACAGGATGATGGTCACAAGGATGCCAAAATTCATGAAGATGGCA
TTGGTTTTGTGTATGGGAGTGGGATCTCGGATTGGATTCGGAGGGCTCCCTCGAA
TCAATCTGAGTTTTCTGAATCTGTTGAATTTGAAAGCTCTATGTTTTCACTGTAATTT
GGGTCTTTTTAATTTCTTCCTATGTAATTTGGGTGTTTCTAATTTCTTCCTTCAGCAA
AAAAAAAAAA SEQ ID NO:9 Middle LPS-012
GGTACTCCACCATATCCAGGTAACAAGGGAAAACAGAGTCAGCTTCTAGTATGTT
GTATGCCTTGCTCTGTCTGTTTTCTTTGATCTTTGATGCCAAGCAAGTTGAATGTGA
TCACTAAATGTTGCTGGCAGTAGAGCTGGAGATGTGCTGTCTCTTTGGTGTCATTA
GCACAGAAGCTATTGGAGAAATGATTATTATCTGTTTGATAACTTCTAGAGCATTTT
TCTGCTTCCAATTCCACAAGGTGGAAAGTGCAAGGATGTTTACTTTCTTAAACTGTA
CTTGCCTTGTATTTGATGATGTAAGGTTGTGTGGCAAAAAAAAAAAA SEQ ID NO:10 Middle
LPS-013 GGTACTCACCATATCCGGTAACAAGGGAACAAGTCAGTTTTA- GAAAGTGGACCCCC
GGTTCCGTCGTTTTCTTGATCTCGGAGCCAAGCAAGTGGA- TGTGATCACTAAATGT
TGCTGGCAGTAGAGGTGGAGATGTGCTGTCTCTTTGGG- TCATTAGCACAGAAGCTA
TTGGAGAAATGATTATGGTATTCCACCATATCCAGG- TAAACAAGGGAAAACAGAGC
TCAGCTTCTAGTATGTTGTATGCCCTGCTCTGTC- TGTTTTCTTTGATCTTTGATGCC
AAGCAAGTTGAATGTGATCACTAAATGTTGC- TGGCAGTAGAGCTGGAGATGTGCTG
TCTCTTTGGTGTCATTAGCACAGAAGCTA- TTGGAGAAATGATTATTATCTGTTTGAT
AACTTCTAGAGCATTTTTCTGCTTCC- AATCCACAAGGTGGAAAGTGCAAGGATGTT
TACTTTCTTAAACTGTACTTGCCT- TGTATTTGATGATGTAAGGTGTGTGGCAAAAA AAAAAAA
SEQ ID NO:11 Middle LPS-014
GGTACTCCACCATATCCATGTAAACAAGGGAAAACAGAGCTCAG- CTTCTAGTATGT
AGTATGCCCTGCTCTGTCTGTTTTCTTGATCTTTGATGCCAA- GCAAGTTGAATGTG
ATCACTAAATGTTGCTGGCAGTAGAGCTGGAGATGTGCTG- TCTCTTTGGTGTCATT
AGCACAGAAGCTATTGGAGAAATGATTATTATCTGTTA- CATAACTTATAGAGCATTTT
TCTGCTTCCAATTCCACAAGGTGGAAAGTGCAAG- GATGTTACTTTCTTAAACTGTA
CTTGCCTTGTATTTGATGATGTAAGGTTGTGT- GGCAAAAAAAAAAAA SEQ ID NO:12 Late
LPS-015 GGTACTCCACTAGACCGGGTAGGGTCTCTCCATGGTTTTGCGACTAGGTTAGGTG
TCCTGTTCTGTTAATGATTTTGAGGTTTTGTTAATTGTGAGTATGTTTCCAGGGTTTT
GAACCTGGGTACTCGGCCTTTGTTGGAATGTAGTCTGGTTAATTTATATGTATATGT
AACCTTGGGGTTTCGAGCCCAGTTCTCTGTTCTTCTTGAAATGAAATGCGATTTGTT
CTAAAAAAAAAAAA SEQ ID NO:13 Late LPS-019
ATATATACGTATGGTATTCCACAGCATGAACTCTCGACATTATATGCTTGTTATAGT
TTTTAAGAGAGGAGACTTACCTCACACATGTACAGCTTTTTATTGTCGTGCTTTCAG
TTGATGGATGATTGTTGTAGTCCTGTCATTGGTTGGACAATTTCATCATCCTAAAG
ATCCAAGAATTCATGTGGCAAGAAACTTTAATAAAGTCAAATATAATCCGATGACGT
AACCCTAAAAAAAAAAAA SEQ ID NO:14 Late LPS-020
GGTACTCCACTAGTGATCGATTCTCTGTATGTGACGCTGCGCGGCGGCTATAGC
GCTTCACTGAGAATGTACGGTATATTATGATTGATGTGATGGATTTGCTCCGCAGC
TTCGGCTGTTGTATCTGCTCACTTCGGCGTATATATGTAATATGTTGCTTCTTCAGA
GAGATGAACTTCCCCCTAAAAAAAAAAAA SEQ ID NO:15 Middle LPS-023
ATAGATCATTTTAAAGTTTCAGTGATTTGAATCTAATTCCACTGCATTTCCTCGCAAA
CTGGCAGTCAAATAGTATTCCCTCTTTCAGTGACAGGCTGGCAGGTGTTCATTCT
TATACAAACATGATTATCATAATTCCATTAATTCATGGCGTTTTCTTTGCCAAAAAAA AAAAA
SEQ ID NO:16 Late LPS-024
TTTTTTTTTTTTAGGGAGAAAGGTAACTTCAGCCAGCTTTCAAAGGCAACACCTACA
AAAGGGGTGACTGAGAACTCAGACACAGACGACAAGTGATCATTCGGGCCAGATT
TTTGTTGAGAGAGTTGTAGTGTGTAATTGATTCATTTCATACATTTGATATGCAAGC
CTGTACAATAGCCTGTGACTGTTAAGGGCATTCTTTTGTCTCCCTGTTGCTATTTGG
GTTTCCGGTGTGTTCATTTTCACTTATTTTTGTGTTTTAGCTGGAAGAATTTGAGAG
GGTAGAATTGTGTCATCGCTATGGCTTGTGCATGACTCATGAGCCAGCAGTTGAAA
CTTTTATTTATTAAGTTATAATACTATGTCTTGTCAATTCTCAATAAAAGATATTTTA- T
GCTGTTGGGCAGCATCTAAAATGTTTTGTATGTTAGCATAAAATCCCATTTTC- TATA
AGTTTTTGCCAAAAAAAAAA SEQ ID NO:17 All LPS-025
AGCAGGTTCAGTCAGACGTGTAAACGACGCCATGATGTATACGAACTCATATAGGG
CGATTGGCCTTTAGATGCATGTTGACGGCCCGCAGTGTGATATTCGCAGATCGCT
TTTTTTTTTTTTAGGCATGGTGCGCGATGAGCTGATAGCGATGATGAAGACCAAGA
CCACCAAAGGAAGATTCTTCAGAGCAAAAGCTACGGAGACAGAACCAGAGGACTC
AAAGCCGGAATCCATTGGTGAGGTACCTGCAAATGTGTGATGGACTAACTAAGA- AG
GCTCCTTGAGAGGACCCATTAAGCACAGTGTTTTAGTCCCAAATTCTGTGCA- AT
TCCGTTGAAAATCATTTTTACGATTTTAGGTATGATGTGTGCAATTTAAAGT- TGGAA
TTATTGTGGGCAAAGGCTATAAGTGATTGTCTAATCCATTTAATTTATT- ATCTTTTGA
CTAAGAGCATATCTAGGCTGGAAGAAATTAGGGCACATATGTTTT- GTGAATTT
GAACATTCTGGGTTTTGCAATGCAAAACACCACAAATATTTTATAA- TGTAGAGGTG
TACTTTTTCTGGCCAAAAAAAAAAAA SEQ ID NO:18 Middle LPS-026
GGTACTCCACCAATAATACTGTCTGTTCTGCTCCCTGCTGATCCA- CTAAGCAGA
TTATTTCTGTCCACCCCACTTTAGAGTCTCAGTTTGTAAAGCACT- CCCTAGGAGCTA
AACTCATTTCCAATGGATTAAAGCACTCCATAGGAGCTAAAC- TCATTTCCAAGGGAT
TTTTGTCCATTTCTCTGTGCTAAAAAAAAAAAA SEQ ID NO:19 Early LPS-027
ATGTATACATATATGTGGTACTCCACACACTCAATA- ACAGCATCACAATCAAAACA
AGAAGGCGGCCAGAAAGCTTTAAAATGCTAAGCC- TACAGGTAATATTCACAACTGC
ATTAAGCACCCCGCTCCTAGTTCTGAAGAAGC- CAGAAAGCTTTAAAATGCTAAGC
CTACAGGTAATATTCACAACTGCATTAAGCA- CCCCGCTTCCTAGTAGGCTAGTACTA
GGACTAGGACCGCATTACCAGTTCCCTT- ATCTTCTACTCATCCTCTACAGGAAAAC
TATGACTAAAACTGCATTACCAGTTC- CCTTATCTTCTCAACTCGTCCTCTACAAAAAA AAAAAA
SEQ ID NO:20 Early LPS-028
GGTAATTTCCACCCACCACGGGCTTTTTCAATTAACCCATTTCTAC- CACTCCACAT
AGGGTTCTAAGTTTTGTGACTCACCCCCAATTTCGCTGATATTT- TGCATTGCAGCT
GTTTATCTACAGGAAATGGCTAATCAGTACTTTCAGAATTGG- TTGCTTCTGTACAG
GAAATGGATAATCAATCAGTACTTCTATACTAAGTTGCTT- ACGCGGGGATCAGAG
CCTTACTTCAGAAAATTGAATACATTTTCTTCTTTGTGT- ATGTATCAGGCATGGAAT
ATATGTAGCATGCCATGGAATGCGTATTTACTAGAT- TATCTTTTAATTTAATACATAT
GTTGCTTACTAATTTGTCCACAAAAAAAAAAA- A SEQ ID NO:21 Early LPS-029
GGTACTCCACACACTCAAACAACAGCATC- ACAATCAAAACAAGAAGGCGGCCAGAA
AGCTTTAAAATGCTAAGCCTACAGGTA- ATATTCACAACTGCATTAAGCACCCCGCTT
CCTAGTTCTGAAGAAGGCCAGAAA- GCTTAAAATGCTAAGCCTACAGGTAATATTCA
CAACTGCATTAAGCACCCCGCTTCCTAGTAGGCTAGTACTAGGACTAGGACCGCAT
TACCAGTTCCCTTATCTTCTACTCATCCTCTACAGGAAAAACTAGGACTAAAACTGC
ATTACCAGTTCCCTTATCTCTCAACTCGTCCTCTACAAAAAAAAAAAA SEQ ID NO:22
Middle LPS-030 GGTACTCCACTATTAGATTGATGCAAGACCAACTGATCATGGCTAGGGT-
GTATCA AGCATTTCCCAGGCTAGGAATAATCTTGATTTATACCATGAATGATGC- TTCGTATT
AAAGAATGTCAACGTACATGGGTGAGACTAATGCCGATCTGATCTA- CCTCAAG
GTAATAATTTTTGCATTAGCTGCTTCTAATCAAGAGTAGTAAGTGCT- TCCATTTGC
AAAAAAAAAAAA SEQ ID NO:23 Middle LPS-031
GGTACTCCACAAGGCATATATGGGCAATTGATTTTGCCTAGCCCAAATCCTATCA
AGCTTGCGTATTTCTAAAAGATGCACTATTTTTTGTCCGAGTGTAGGTTTTGAATTC
ATTGTAACATTCAGCAATATTAATTCAGGGGTAGCATTTCTGGCAAAAAAAAAA- AA SEQ ID
NO:24 Middle LPS-032 TTTTTTTTTTTTAGGGTAGAAAACCAT-
GCTTCACTAACAAGGTATTAAAATTACAATAT
AATTCTGGGTGTAAACGACCTGATAGATGATCTGCAAGTGCCAGGAGGCAATATCT
AGCAGAATACGTACAAATAAATTGCCAAAAAAAAAAA SEQ ID NO:25 Late LPS-036
GGTACTCCACCAATGATCACCCATGTCCATTTGGTTAATTCAATGTCAAGATTTAGT
AGTTCCGTATTCCCTTGGGTAAGCTGTAATGGTCCATTTGGGAACAGTCCATGTTT
GGGACACAAGTTCAATAGAGATGTCATCCATAAATATCCATAAATATGGGTATGAATCTC-
TTCCTC CCTCTCCGCCCAATAATAAAAAAAAAA SEQ ID NO:26 Late LPS-037
TTTTTTTTTTTTAGTAGCAATAGCAATCCATTTTAGGGATCTGCAGATCAG- TGACTAA
GTGACCCCTACCCCCAAAGGATTAATTGTACTTTGGCTTAACCACAA- AACCTGATC
AATGTGAAGTTTTTACCCATATTAATTCCCAAAAGTAACTACAAA- TTCCAG
AGTACATTTTTACCCAAAAAAAAAAA SEQ ID NO:27 Middle LPS-038
GGTACTCCACTATACAATATCAAGGCATATCTGCCGGTGTGAATCATTC- GGATC
TCAAGCACTCTCCGTGCCGCAACTTCTGGCCAGGCTTTCCCTCAATGTG- TGTTTGA
CCACTGGGATATGATGGGATCTGATCCATGGAACCTGGTCCCAAGCT- GGGCAG
CTTGTGACTGATATCCGTAAGAGGAAGGGTCTTAAGGAGAGTATGACT- CCCTGTC
AGAGTTCGAAGACAAGCTGTAGAGCTTTGCTATGTTTGCATGTCGGA- TGCTGTCAA
GATTGAGGAACCTCCGAGTATTAACACAGTTTTGTGTGCTAGGAC- TAAATT
TATGCTATTCACGTATTTTTGTGATCTGTATTTATGTTATCACGTATT- TTTGATTG
GAAAATACTTTTTACAAGTCATCCATTAATCTTTTAAATGTTACAT- AATTCTCTCTGT C SEQ
ID NO:28 Late LPS-040
AAGCTTGGTACCGAGCTCGGATCCACTAGTAACGGCCGCCAGTGTGCTGGAATC
GGCTTGGTACTCCACTATACAACATCAAGGCATATCTG SEQ ID NO:29 M,L LPS-041
CTTTTCTTCGTGCTTTCGTGGAGTACC SEQ ID NO:30 Middle LPS-042
GGTACTCCACAAAGTGAGATGAGTGATATGAGGTCAAACACGTAAATGACAA- TAGC
TATTATTTCCCCACTTGTTTGTGGCTGTGTATATTATACTTCATTGTCAG- GACTTTTG
TATGGTTGAAGTTGCAAGGTTTTGGCAAAAAAAAAAAAAAAA SEQ ID NO:31 Middle
LPS-043 GGTACTCCACCTCCAGCTGCTTATCCAAGTACT- ACGGATAGTTCATACTCCTATTAT
GCTTCTGCCAAGTGAACCAGAAGGCTTCTG- TTCTACACTAGCAAACTGATAGCTC
GAGCATCTCATTTACTAAGGATGATAATT- CAAAATTGTAACATTGCAAACATCAGC
AAACATCAGCATCAACTCTGTTACTAT- ACAAGCAATGGATGCGTCGCTGATGCTG
CGGGAGAGTAAATTTTTAGTTTACTG- CGGTTGGTAATTGAGTAGGTTGACTTACATT
TCTGTTGTAAAGCCGTTGTCGGG- CATTGTTTATCTGGCCGAGTTAGCGCCAGGAAG
CTAAATGTACCAAATATTTATTATTTTTATTAAGAATATAAAATTTAGTCGTCTTCT
GCTGCCCAAAAAAAAAAAAAAAA SEQ ID NO:32 Late LPS-044
ATGGCCATGGACTTATGACTTTCAAAACCCTAAAACCTATCTACAACTTTCCACGCT
GAGATTTTCCGAGGAAGGCATTCTAAGCCATTCCCACCGTACTTTAATAAAATAAAA
ACAAGAAGATAGTAAAGCTAAGCTACAACCTTCCGCCAAAAAAAAAAAA SEQ ID NO:33 Late
LPS-045 GACCGCTGTAGGAACACTAGCAGATTCCGGAACATAGGTACTTTGAA- CATCTTTC
ACTCCTCACCATATGAATAGTGAGTCGATGGCGGCCTTAACAGTCG- AGCATGCTTT
GATTTCGTCTCTCTCTCTAGTGACCGGAAATCAATCTCATTATA- TATGTCATTATGCAT
TCATTCCCACTTCCTAACTTTCATTATTGTTCAAAACTC- GCCTTCCTGAAAATGCTA
TAATAGTAGGGGAATATTGAAAAACTTCCGCCAAGC- TAAAAAGGCACTTAAAGCAC
CTGGATTTGAACGAGGATTTCCCACCCCGATGAG- GGGGGGTGTCTTTCCATTGAG
ACGATGCCTTACTCGGCAGACCCTGTGGGGGTC- TTTATAGGTGACTTAATACTTAA
GTATAGGACTAAGAGAGAGGAAGCGACCGCC- TCTCTGATCAAGCCTTTACGTGC
GACGTGCCCAGGTAAAGGCTGATCTCACCAA- ATAATTCAGAGAAAGAAGATGACTC
CACAGTAGCGAAACTCCTACATTGTCTTA- CATATCGTAACAAGCGGTC SEQ ID NO:34
Middle LPS-046
GACCGCTTGTGCGTGGTGTCCAAACTAGGACGCCTTAGTTTTCCTAAGAAGGAAAC
CCAGGCGTTGACTTGAGGCAGACTTGTGCTTCTGGGTACTCTCATCACTGCGTGA
CCTTGAGAAAGGGACTTTACCTCCAGGATCCTCAAACTTCTTCTCTGTAAAATGAGC
ATTGTAATAATTATATCCCAGGCTTATGTTGGGAATATTCAATAAATGCTCCCTTCAT
TCTTTAAAAAATAAGTAAAGACAGCCTGAATGGGAGCCACGTTCTCATTCTTCTTTC
TCTTTAAAAAATAAGTAAAGACAGCCTGAATGGGAGCCACGTTCTCATCTTCTTTC
TCTATGCAAAATGTATTGTGTAATGTTTGTGTACTAGTAGTTCAAGAGCAAATAAGT
AGTTGGTTAATGGCTAACATATTTCTTAAATTTGTAACTGTTAAGATAAACATT- GAAC
AAGGAAAAAGATTCGTAACTGAAATGTAAAGTCATTTGACCCTGGATAGT- CAATGAC
AATCTTATTCACAGTGTAATAAGTAATTCATAACGAGATGATTATTA- TGAAATTATCA
ATAGCCTGCTATATCACTTTATGTTTATGATCCACAAGCGGTC SEQ ID NO:35 All
LPS-047 GACCGCTTGTGGAAGAAAAGAAAGAATCTCTT- TCGGATTCAATAGGCGGTATGGGA
GAGTCTGCTACTGCCTCTTGGATTCCAGGA- ATCCTAGAGCTGGGAGTATGAGTTGG
AGATGATGAAGGTGTCTCTTACCTATTT- CTTGAAGTGGATGGAGTTGTGAAAATCG
ACTTCTAGCTTCAGCTAAAAACCTTC- CCCTAGAATCTCTTGCTCTATGCATATCATTT
TTATTTTTTCTTTCAAGATAGGGTAATAATTCTCTTTCTGATCTTCCAGGTCACTCTA
GGTGCAAGAAGAGAGCATAGTCAAGGAACTATTAAACCAATAACTTTCTCTTTTCTG
ATCCTCCAGTTCACTCTAGGTACAAGCGGTC SEQ ID NO:36 All LPS-050
GACCGCTTGTGCAAAGTAGATACCGTCCTGTTCCGGTGAATTGAAGTACATTTTCA
AAATGCGCTACTATGACATTTTATAGGATGTCTGAGTGTAAAATAATGGTACTGGTT
GTTGCAAAGAATCTGATGTTTGGATGTATGGAACTATAAATAGATGTTATTTTCTGA
TCCAGAAGGCTTTCCTTACCAACTGATTTCATCTCAGAAACTAAAAGCTCTTGAAC
TTGTGTAGATGGGGCTTGGTCATTGTAGTTTAAATGCATTATGTAGTGGCAAAA- AAA
AAAAGTTATAGCCTACGTTCAAATGGATTTGCTCGACAATCAAATGAATTA- CAATT
GAATATTCATGTATACCCAAATTTTAAATGTAGAATGACATCATCAATG- TAGACAAAC
ACCACTGTGCTTGTCCTTGATATCCTCTTTCACCATATAATTGGT- GGCTACTCAAA
GTCACTATCTGATGCAACTACAAGCGGTC SEQ ID NO:37 Late LPS-051
GACCGCTTGTTCAATGCAGAATCTCGAAGAGATGTCTTGGACAAA-
TACTGAACTGG CACGATTGGTGTAGTGCGGTTCAAAAGGCGCTCCAGATTCGTC-
TGGAACGAATCTT CATACGCTGAACAATTAGACATCTTGTACGCAAGAGAATTA-
CGATCGGCCATATAAA AACCCCAAAGAGAAGAAAGTGTTTCGAAATTCTCCCAG-
AAAACAGTCTTATGCCAC CGATTTGTCTTTTCAACATGCATTTGCAATGAAGTC-
TTTGGATTCTTACTGTGAGTG CTGATCAGCAACGGATTTTCGATCTGTATAGCT-
CTGCCGATTCCTGGTTAAAGCAG CTAAGAGTTAGGCATCCAGATTTGAGTTTTT-
TGCATCTCACAATGTTTGAATACAT TCAAATCCATTGTTGGAGTAACCTAACAA-
CAACTGTACTCTTCTTCCTATTTCTGAA GCCCTCTGCCAGTTTAAGGCAGAGAA-
CTGAGTATCTACAAGCGGTC SEQ ID NO:38 Late LPS-052
GACCGCTTGTATAATAAAGTGGTACCGCGTCCTGCAAACAGGGTTCTCTTGCCATC
CTGCTACAACCCTGCAGTGGTCGCAGTAGAGAGAATCGGAGCAACGAACGTTTTC
CCGAATATATGGAGCGGGAGGAAGAGTTTTCTTGCTGATGATCCAATCGGAGTCGA
ACTGCCACCGCTGGATGAAGGGCGGCGAGGAAATCTTGGGGGGCAGAGGCCCGT
CGGCGTAGGAAATAAGAAACGATTTGATATGGAACGAAAGGGCCCGTCCAGGGTT
CGATCCCCGGCAGGGCAGCCAGCCCCGAACTAAACAAAACAATAAGAACAAACAG
CAAAGTAAAAGAAAGCACCAGAAGAAACAGCAGCAGACGAAGAGTAAGGAGCTGC
CCACAAGCGGTC SEQ ID NO:39 All LPS-053
GACCGCTTGTAATCCACAGCATTTTCAATAACTTCCTGAGGTGACATCCACCTCCAC
TCAGAAAACTCGGCTGCATCTGTCCCATCACCAGCTAGATTGATCTCACTCTCGTC
TCCTCTAAATTTTAGGAGGAACCATTTCTGTGCTTGACCTTTCCATTCGCCTCCCCA CAAGCGGTC
SEQ ID NO:40 Middle LPS-054
GACCGCTTGTATATAATGTGAAGACACAATAAAATTTTGTCCAACAAAGCAACCAAA
CGACCAAAAATTTAGCTGTGACATCAAAAAGCTCAACCCCTACAATGAATGTAACCT
TAATCTAGAAAATTGATCCATGATCTCCACTGAATTTCTCGTCATCCTGAAGAAT
GAGAAACTTAAATGTACCCGATTCCCTCAACCAAGCCCCCACAAGCGGTC SEQ ID NO:41
Early LPS-055 GACCGCTTGTAATCCACAGCATTTTCAATAACTTCCTGAGGTGACA-
TCCACCTCCAC TCAGAAAACTCGGCTGCATCTGTCCCATCACCAGCTAGATTGA-
TCTCACTCTCGTC TCCTCTAAATTTTAGGAGGAACCTGTGATTGGTAGGGGCTT-
GTCATAAATGATCAAG ACGACCCGCATCGTGATGCCAAGCTTAGTCTTTCTACT-
TACTGTCTATGTAATGGTC ACGGGCCCTTCTTATGTTTATGTCTCTTTGAAATG-
GACGATTTTTTTGTTTTAGGTAT ACGGGCCCTTCTTATGTTTATGTCTCTTTGA-
AATGGACGATTTTTTTGTTAGGTAT TCAGTTTCTGAAGCTGTTTTGGTAGTAAA-
CTGGGCTCAATCATTTCTGTTGCTGAA CTTTCCATTCGCCTCCCCCACAAGCGT-
CAGCCGAATTCTGCAGATATCCATCACCT GGGGGGGCCGCTCGAACATGCATCT-
AGAAGGCCAATCCCCTATATGAATTCTATTA AATCCCTGGCCTCGTTTTA SEQ ID NO:42
Early LPS-056 GGTGCGATCCAGAACTATCATCTCTCACTGCTCG-
TGAACAAAATGCTGGTCAT AGCCATCACTAAGGCTAAGGTACTATCCAGCCAAA-
CTGATCTCAAATAATAATTTCA TAAGCTTAAATAAATAGTCCAGCCAGTAGATG-
GAGCCAAAAAGCCATAGAAGCTC AAATACTTGTGGTATCAATCTCTCCTCTGTT-
AAGGGAGGTATCAGATCAGAAGCACT AATCAAATGCATACATAAATGCAGTAGA-
CTGCAATAAAACAAAATCTGCAGATAGCA ACAGAGCGCTTAACGAACGGAAAAG-
AGTTTAACTTGATCTATCACAGGATCGCACC SEQ ID NO:43 All LPS-057
GGTGCGATCCACAATAGTTCGTACGAGCGACGTCTATCTGGTTAATCAGAACACAT
ATCTAATTTGGAAATTTGTGGGCATAAAGCTCCACAGTGTAGGTGGGCTAATCCCA
TGAAACATTACTCTTCAAACATCATACAACTGAGGTGGAAATGCAAAAGATTATT
ACTGGATGCTGATCTGGGACTAAGGTGGTGGCCATTGGTAATGTGTGTTTCAGAA
ATATATCTTCATGATGATCAGTAGTTGCATCTGGTTGGAAGAATGATAAATTCTGGT
AATTTGTCTTGGGATCGCACC SEQ ID NO:44 Late LPS-058
GGTGCGATCCAACTAGAAGAATATAAAGAAAAATTACGGACTACCAGAAAACATCA
CATCACAGTGTATGCATTCTCAATAATCAGAACTGTACTGGCTAATATCGCTGTGC
CTGTCGTTTCATTTTCCTGTCATCCGCATAGGGCCCCTCATTTTCCCTATCTTGCAG
AAATCCAAGAAATGCAAGAAAACCAAAAAGGAAGAAACCCCCAGAGGAAGAGTCCG
AAGAGGATATGGGTGTCAGTCTTTTTGACTAGATTGGAGGATCGCACC SEQ ID NO:45 Early
LPS-059 GGTGCGATCCCAGAACATTTCAGACAGATTAAAACAAGATCTAGT- CAATCCTACAA
GGGAAACTTTTGTCAAGATCCGGATCCAGATTTTCCTCAAGTA- AAACTAATCTCATT
AAATCCAAGCCAATCTCTAGCAAAATTCAAACACTTTTTA- TTAAATCCAAGCCATATA
TCTGGCAAATTCACCGAAATATGTACAATCGCAGCG- CATTGCTTGGCTTGCGACAG
AAACCATATTCGCACGTCTTCATAAGGCTTTGGA- TCGCACC SEQ ID NO:46 All
LPS-060 GGTGCGATCCAACAACACAGCTTCA- CACTTACTCCATCCTCTGGAACTCTCATCAG
ATTGTGTTCTTCGTAGACCAAGT- TCCTGTGAGAGTCCACAGGCACACTGAGGCTAC
AAGCGATGTGTTCCCTAAAGAACAGGGGATGTACATGTTTTCCAGCATTTGGAATG
CAGACGACTGGGCAACCAGGGGTGGGCTTGGGAAGACAAACTGGACTGCCGCTC
CATTCAGCGGATCGCACC SEQ ID NO:47 All LPS-061
GGTGCGATCCCAACACCAAGTGAGAATGAAGCAATATAAATCAGCAGACTCACTAA
AGCCAAAACAGTGAAAAATGTTTCATATTGGGAATCTGCTCCAGAATGAGCCTTCAA
GTAAAATGACAAACTAACGAGGAAGAGACATACGGCCATGCCCCCAGATGAGACC
ATGAGGAGGAGACGTCGTCCGGCTTTATCCATGAGCCATACAGCAACTGCAGTCAT
GATGACCTGGATCGCACC SEQ ID NO:48 Late LPS-062
GGTGCGATCCAGGAAATCATCAAAGGGGAGCACATCCAATGTGCAAAATAAGATCA
TCATGCAGCAAGATCTCTGAAATATAAGCTCTGTAAGACCAATCTGAAGTGCTGATG
ATCAATATGAACTGAAACATCATGCCACAATGGGCTGGTACTTGTGCAAAATTCTCT
GGCATGTGATGAGAATCACATGGTTACCTCTTTGGATCGCACC SEQ ID NO:49 Early
LPS-063 GGTGCGATCCAAAGAGCCTTCTTGCAGACAATCCGTGAAAACATGGCTAT- ACAATA
AATTCCCAGTTTGGAATTCTAAATAAAACTGTTCAATATTTGAAGGCC- TCTGATATCA
CAGAGACTGATATTAGAATGGAAGCATGTAGCAACCCTAGAAGC- TTTCGCATAAAG
ATACCAGATTAATTCATAAGAAGGATCTCTCGTTCACCAGTC- ACATATCACAGTCGG
ATCGCACC SEQ ID NO:50 Late LPS-064
GGTGCGATCCGTTAGATGAGCTGCCAAGTATGGAATTATTGACATTTTTGGACGGG
TTATGGGCAGAGGGATGTGCCAAGCTGAAGAAGATACCGGGGTTGGAGCAAGCCA
CAAAACTTCGAGAGTTAGATGTTAGTGGGTGCCCTCAGTTAGATGAGCTGCCAAGT
ATGGAATTATTGACATCTTTGGACGGCTTGTGGGCAAAGGGATCGCACC SEQ ID NO:51
Middle LPS-065 GGTGCGATCCACATAGTTTGAATGCAAGGAAATT-
GCACATACTTCGTGGGGAATTT CGATGGCAAATCAGTCCAGGTAAATGACTTCT-
CAACATAGGTCCAAAACTCTTTCAT AGACCAGATCTTGACCGTGTTGTCCATGC-
CACAGCTGCAATACGATATACATCTG AAGGATGAAAATCTACACTGAGAACTTC-
ATTGCGATGTCCCCCAGCTCCAGCAAAT ATCAAAATGCATATTCCAGTTTGAAC-
ATTCCAGAGTCGTACAGATTCATCTTTGCTA GCAGATAAAATAAGGGAAGGTTT-
CAGTTGCTTGGGTCCTTATTTCATTCACAGAACT
CCATGGCCAACGAAACTCTTATGGACTTTTCATTTGCACATCCATTCTCGAATTATA
CATTGTGACCGCAGCCACTAATAATGGGGAACATCACTCGCCTGCCGACTTATGTG TTAAAGAATC
SEQ ID NO:52 Late LPS-066
GGTGCGATCCCCTCCATTTACCATGGTATACTGTTCCAAAGGTTCCAGAGCCTAGC
TCTTTCAATTCTTCAAGGTCAGCATTCTTTATATCTGGAAACTTCGCTAGCTGTGT
CTATAATCACGAAACCCAGACGGGGAACTAATAGGCGATGAAGTTCTCTTATCCA
TAACCGTTGCAAAGATCTTACACGGAGTTTTCTCTTCTTCTGCGTGGCTTTtCTTTC
CCGTATTCTCGGATCGCACC SEQ ID NO:53 Late LPS-067
GGTGCGATCCATACATGCGAGGGCGCATGAGAGACTACCACAAATCCTACATACCT
CCATTCACCCCTGGATCGGTTATACAAGGATTTGGGGTGGCTAAAGTGATACTCTC
AAATCACCCAGACTTCAGAGAGGGTGACTTGTATCTGGTACTATAGGATGGGAAG
AGTACAGCATAATACCAAAAGGGAGTAACTTAAGAAAGATCAAATATACGGACGTAC
CACTTTCATATTTTGTGGGTGTTTTAAGAATGCCCGGGTTTACTGCTTATGCTGGAT
TCTTTGAAGTTTGCTCTCCTAAAAAGGGGGAGCATGTTTTTGTCTCTGCCGCTTCA
GGAGCTGTTGGCCAGCTTGTTGGGCACTTTGCAAAGTTGATGGGTTGCTATGTTTGT
TAGGGAGCGCGGGTAACAAACAGAAGGCTGATCTGCTGAAACATAAAATGGGCTTT
GATGATGATCTCCACCATAACGAGGAGCATGACTTCGATGTGGCTTTAAAAAGG- CA
TTTTCCAGATGGGATTGCACC SEQ ID NO:54 Late LPS-069
GGTGCGATCGAACTGAATGAATGAOGTTGCCAAGCTATGTTTGGGAATTAAAACTT
GAATGCCGTTATTCTCTCCTTTTTCCAAAAGGGCCTTTTCTGCCAGAAAACCTTAAA
TTTCTGACTGGTTTCCAAGTCCAATTTTTCCAAATATGGATTGGTTTACCATTG- AAGG
CACCACCATGCTCTGAAAGTTATGGACTGCACTTGCCCCAGTGCTATATT- TAGTCC
AGATAGCGCTTGTGTCTCTAAATGCATCTCCCTGCTCGGATATCACC SEQ ID NO:55 Late
LPS-070 GGTGCGATCCGAACAGAGGGAGCAGATTTTG- CCCTGCAAGTATTCACAACATTAG
AGAAGCCCTGCCAGAGATATGGGAGGAAGA- AGATGCAGAGAACACCAAAAATGTT
GTGGGATCAAGAGGAGCGGATGCAACTAT- AGAAACTGTTGTCACGGCATAAGCCA
TCGCCTCATTGAATGAGGGAATGGAGGA- CTAGACAAATCCCTTTGGATCGCACC SEQ ID
NO:56 Middle LPS-071
GGTGCGATCCGATTGGGCAGCTGCAGCCTTGGGAAGCTTTAGAATCAAATTGCAC
TCATCCTCCAGGAGGTATTGAGAAGTCAATTTCTCAAGGTCTACAGTGACAGAAGG
AACCATCTTGACAATCTTATCAGGTTTCCTGCTCTGGTTAAACACTTCAACTTTGAC
AGGACGAGAGTATGTGACTAATTCATCTTCTTCATCAGACTCTACATCTCCTGTTT
CAAGAAACAAAGATACTGATCATCACTAGGGCAAGAATTGATGATTTTGATATCTCT
GGAGAAGCCAGTGTTTACATTGGTTTGCTTCATGGCCACCAGTCTATGGCATAAAG
CTTTCCCGAAAGGGTACTTGGCAGATTTAACAGAGCCCAACGTTATATTTAAGGCC
CATCTCTTTGCTCTCAAAATTTTTCTTGCATCCTCTGGAGAATATAAAACCCCTTGG
TGTCTCTTTCCACAAACACCTTCTCATTGATC SEQ ID NO:57 Late LPS-072
GGTGCGATCCAACTGAGAGGGTGTTGGTGGAAAGATGACACCAAGTGGGTT- CT
ATATTCTCCAGAGGATGCAAGAAAAATTTTGAGAGAAAGAAGATGGGCCCTT- AAAT
TAACGTGGGGTTCTGTTAAATCTGCCAAGTACCCTTCAGGAAAGTTTATG- CCATAG
ACTTGGTGGCCATGAAGCAAACCAATGTAAACACTGGTTCTCCAGAGA- TATCAAAA
TCATCAATTCTTGCCCTAGTGATGATCAGGAAGATGTAGAGTCTGA- TGAAGAAGAT
GAATTAGTCACATTCTCTCGTCCTGTCAAAGTTGAAGTGCTTAA- CCAGAGCAGGAA
ACCTGATAAGATTGTCAAGATGGTtCCTTCTGTCACTGTAGA- CCTTGAGAAATGAC
TTCTCAATACCTCCTGGAGGATGAGTGCAATTTGATTCTA- AAGCTTCCCAAGGCTG
CAGCTGCCCAATCGGATCGCACC SEQ ID NO:58 Late LPS-073
GGTGCGATCCATGTAGTGCCAACTTACGAGATCACTAACTTTAAAA- CTATCATGCAA
TTGGCCAATAGAAGCGACACTTGCTGTGCCAAAGTATCGATAG- GCTACTCCCGATG
GCTCAATCATATATAGTTGGGGCCCATCTCTATCATAACCT- CCAAGGATAACTCCAG
ATCCAAAAGGCCTTAACCACCAATATAGTGTGCACAAA- TGCACATAACTGGCAACA
CGTTCACAAAGTTCCTTAAT SEQ ID NO:59 All LPS-074
GGTGCGATCCCATGGGATAGTTGCAAGACACACAAATTTGTTGTGAAA- GAAGAGAG
ACACGCACAGACAACCATATGATCTTTTTTTTTTTTTTTTTTTTTT- TTTTTTTTTTTTTT
TTAGCAAAATTCAAACACTTTTTATTAAATCCAAGCCATA- TATCTGGCAAATTCACCG
AAATATGTACAATCGCAGCGCATTGCTTGGCTTGCG- ACAGAAACCATATTCGCACG
TCTTCATAAGGCTTTGGATCGCACC SEQ ID NO:60 Early LPS-075
GGTGCGATCCCACTGTAGTGTCCTTGTTGAGCATAGTTC- AAGCTGTTCTGATTCC
ACCAGTTAGTGGCCCAACACTGCGAGGTGCTGCCATTT- CCATTCCATTCACAGACG
TCAGTGTTGAAATTCATATAGGAAGCCACAAAGGGT- GAGGAAGACCAATCTATTTT
ACTCGCCCCCCTTGAGTTGCCCACTGGTCTCCGC- TCCATATGCTAGAGAATACTCT
CATTGCCTGCTCATTCGGATAGGGAACGCCTA- TGTTTTCATTGTTTGCAAATACTCT
GATTGGCAAACCATCAACGAAAATCGCAA- TTTGCTGGGGGTTCCAGAGAATAGAGT
AATTGTGGAAATCTGCTGTAGGATCGC- ACC SEQ ID NO:61 Early LPS-076
GGTGCGATCCCACACTCCTAACCCTAT- TATATGTCTCCCGTCCATGGAGTCATAGA
AGGAGTACGATAATATGCCCTTCAG- CCAAGCGAAGTATGACTTTAGTATGGCCAGG
CAGCAGTATGAAAGCACATCTTG- TTTCTTCCAGGTCGGCATGTATAGTCTCCGGAG
GCTAACAATGTCACCCAAAGCTAATTGCGCAAACGGAACTCCTCTGCTGATCTCCC
GGGAACTTAGGCGGAACCACCCTGAATCCACTATTCTCACCGCGCATTTCATCCCT
TTGGTGAACGCCGCTGCCTCTGGTAGATACAGAGCTGGCTTGTCTCCACTGGAAC
CCCCTTTCCGGATCGCACC SEQ ID NO:62 All LPS-077
GGTGCGATCCAAACTGTGGTTATCGGTGGAGAGATTAAGCAATTTATTGGAGTAGC
AAGTACGCTGAATTAAGGGGGTCCATCTTCAAGCAAAGGTTCCTTTGGATGACTAT
GTGTTCTGGAAGTGTTTATGGATCAATCATCTCATAAATTTTGGTAATATATAACAGA
AGATTATGGCATCCAGTTAGGATGGTAGTTTCATTGAGGTATAGTAAAAACTACACT
AGTCTTGTGTTGCCACCCACTTTTCAGAGAAGTCAGGAGGTCTCTTTGTGAATCATT
GATAACTTTATGAGTGGGTACCTAAATGAAATATTTGCATCTTGAGTATATACTCAAT
TGATCTTACTTGTGGATCGCAC SEQ ID NO:63 Middle LPS-078
CTTGGTACCGAGCTCGGATCCACTAGTAACGGCCGCCAGTGTGCTGGAATTTACG
GCTGCGAGAAGACGACAGAACACCTATCATAACTTGAATTCTGATGCAAATCGGAA
TTTGCCAAAAACTTGGACGGAAATATAATAGGCAATATCATCCCCGCAAGTAAC- AAA
AAAATTGCATGAAAGCTCAAATCCTATGTGCTTTACACCTTGACTGCATAC- TTTCTC
ATTGGAAAATACATCTCTTTCTTTTTCTGTCTCTCAGTCTTCAATGAC- GGCTGATGC
TGGTAAGGCGTCGCCTGATAGCACGAGTCTTCTTGGGACGCAAAT- CAAGAGGCAG
GTACTTCTTTTTTTTGTATGCTTCTCTTAATGCGGATCGCACC SEQ ID NO:64 Late
LPS-079 GGTGCGATCCAAGATTGTACGGCACAGGCAA- ATGCTGTTCTTTTTCTTAATCACGA
TGTGCTTGAAGAATATGAGCGCCGATGTG- AACAGATCCACAACCTGGAGTTAAAAT
TGGAGGAAGACAGAGCAGTGCTGAATA- GGAGCTTGGCAGAAATAAATAGTCTTTAAG
GAATCCTGGCTTCCCACATTGAGG- AGTTTGGTTACCAGAATTAATGAAACTTTCAGC
CACAACTTTCAAGGGATGGCTGTTGCTGGAGAAGTTACACTAGATGAACATGGCAT
GGATTTTGACAAGTTATGGTATTCTAATAAAAGTCAAGTCAGGCAAACTGGACAGT
TGCAGGTATTGAATTGCTCATCATCAGTCTGGAGGGATCGCACC SEQ ID NO:65 All
LPS-080 GGTGCGATCCGAGGGAAGCGATGTAGTCTTGCCCCAAGCGACGACCATGATCCCT
TATTCTTGGGCAATATGTGCAAGACGTGGACAAATGAAGCGGTTAAAGGGAAGC- TT
ATGGACTATGGAATAGAGGGTCTTGAAGAGCTAACTCTAGTGGGTGATACTC- AAAA
TGAAGGAATAAGCCGTGGTTTTGCATTTATAGCATTTCTACGCACATGGA- TGCGAT
GAATGCATACAAACGCCTTCAGAGGCCAGATGTTATTTTTGGTGCTGA- TCGAACTG
CGAATGTGGCATTTGCAGAGCCACTGCGTGAGCCTGACGAAGAGAT- CATGGCCCA
GGTTAAGTCAGTGTTGTTGATGGGATCGCACC SEQ ID NO:66 Late LPS-081
GGTGCGATCCAGTCCTGAAAATGTACTTTACCATTTGTATAATG- ATGTAAAAATCTT
GGCCATAGTCTGGTCAAACCAGACTGTATTGTTGCTAAAGT- ATGGAAATTCTGGC
CATATTTTTGTCTAACCAGACTGTATTGTTGCCAAAGTTA- TGGGAATTCCGGCTATA
TTTTTGTCTTCGAAAAAAAAAAAAAAAAAAAAAAAAA- AAAAAAAAAAAGATCATAGGG
TTGTCTGTGCGTGTCTCTCTTCTTACACAACAA- ATTTGTGTGTTTTGCAACTATCCC
ATGGGATCGCACC SEQ ID NO:67 Early LPS-083
GGTGCGATCCGCTGGAAGGTGGGCAGCTGGACATCTGGGAATTATA- AGTCGAATG
TCAATTGCTGGGCCATCTGGGGGATGAGCAATAGCATCGGAGGCC- AAGTTCTTCT
GCAGCCGGGCACCAAATGCCATGTGGAGGTCTGAATCTTAGTTT- GGAGGTCGAAG
TTTCAATCCCCTTGTGTTTACTCTGTTTCTGGTTTTATTTGAA- TAATTTGAGCAATTT
AATGTGGGTCCTTAGTGCTTCTGTGGATCAGATTCTAGG- GAACGCCATCCTGATAA
GTAAAGATCCGAGTTTTAATGGAGATTCAATCTATCA- GAATTCCATGGTGGTTTAA
ATTCCCTTGTACTGTTGATCTACGTCGCTTTGTAT- ATCAGTGTGTGTTAAGATTTTCT
CAGAATCCACAGCTTTGTTATGGATCGCACC SEQ ID NO:68 Middle LPS-084
GGTGCGATCCAAGCACTTACGACTCCCAA- CAAGGACGGGAAACTCTAAAATCGG
AAATATCATATACTGAGGCATCAACTTTG- TTGATAAAACTTTAAACAAGAACAATATT
TGCAGCATATTAGCCCACATGCCAT- AATGACAAACAAATATGAGAACACTGCCTACA
GGTTTGCCAAAAGCATGGCCCTCACTTTTGCCCTGAGGTCATCAGGAGCTTCTGAG
GCTCGAGAAGGAGAAAAAGATTGTGTCACTTCAGGAGCTGAGGCCTCCACATCTTT SEQ ID
NO:69 Early LPS-086 GGTGCGATCCAAGGTACGAGCGAACAAGTTTCTTCAGCAAGCCAC-
CTGGAACTTTC CATGAGTCCAAAACAAGTTGAAGAAGGCTTCTTTGGCTACTTT-
TAAGATGCTGAAGT GATTGTGCTCGCCTCTTGCACAGTTCAACCGCAATAACAT-
TGGGTTTTACAAAACC GATTACCTGTTTAACCTGCTGTGCACTCTTTTTCGAAA-
CATGACAAGTTCCAACAAG ATAAACTTCGGCCCCATTCTCGCCATTCCGCAAAT-
AAACCACGCTCTCATCTTCTGT TATCGAACTCGAGTGCATGCCACGACGCTCAA-
TTGCAGGATTCCAACCCCGGACTT GCGAATGGTGCAAAGCGATGCCCGTTTCGT-
CTCAGCGATACTGCTAAAGATCGGCA GACCCGAACCAGTTTGATGCTTCCATTG-
CCTTAAACATCCAGAGTTTTCCTTCGACC TTAAACCCTAACAAGATTACTGATT-
TCTGGTCCGGATGTTCACTGTCTGTTATACTT
CTCACAAATCTGTCACACTCCTGATAATCTTCGGTATTGAACTTCATTGAATTGAATT
TTCCTTCTCATTGGAATTCAATTGTACCTTGTAAATGTCTGGATCCTACACTATACCA
ATATTTACAGGTCTGAGTATTTTGCCTGTAGTATAATTATCTTTCCTTCGGTCTCGT
GTTTCCGTATTATTCGTGTAGGATCGCACC SEQ ID NO:70 Late LPS-087
GGTGCGATCCCGGGGGGAGGTTGATGTTCTGAGAGAATCAATGAAGGGATTTCAG
CTGAGCTTGCCTTTTTGAAGACGGAATGCGAACAACCAGTCATTTGCAATAGCGAG
AATTCTCTTAAGCCACTGCCTGCTGGGGAGGCGAGTTCTGATTCCGGTGATTGCAT
CACTCAACGGCAGCAGCAGCGGCAGAACCTTTAGTTTCCCATGACAGGTCTCTC- TG
TACAAGTATCTTCCTGTATGATCTAATCCGGGTTGTTCGATTATCGTGATGT- CTC
CTGTATTGACATATTAGCAGAATATTACCATGATACGATGTTAAGTGGCAT- GGTTTA
TGCCCTGCATGTTATGTTATGGAGGAGGTGAGGCATGTGGCGCTCATG- GGAGGGC
CCACATGGTCCATGGACGTCTTATTAAACGCATAGTCGTGAATGAAA- ATAGTTCAAT
ACATTCAAAATTCCAACACAATTTCATTACAATGGAAGTGACTT- CGACTTGAATGTT
CATTGAAGCATTTGCATGCACAAACAAAGTATACTAGATTA- GAAGAAAATTGCAAAA
AAGGACATTGTGCCCTTCTTAGTGAATATATAAAGATG- TTCTTCATGCTGGATCGCA CC SEQ
ID NO:71 Middle LPS-088
GGTGCGATCCCAATAGCCAATATTGCCTCCAAGATAGCCTAGACTGCCTTTGCAT
AGTTCTAGAAGCCAGTCACCCAACCTCCCAAAAGAAATTGCGCAATCTTTCCCATC
AGTTTCCCGGGTATGTGTTCTGTCATTCCCCGAATTTTCTTTGGTTTCACTAAT- AG
ATTTCTTTCCATGCACATTGCTTGTCTCCAGATCTTTTAGGTGTTCATCCAT- CTCTTA
GTAGTACTAGATCGATGGCTTCCAAGAGAACAGGATCATATGACACTG- TTGGAAAT
GTAGCTGGAGCAGCAGTTGAGCAAGTGTCCTCTAGTCTATCTATCT- ATGAAAGATA
CACATTGTTTCTAGACATGGATATCAAATTGAAATTGCCAGAAG- TCCATGAAACATT
TGCCGCCTTTTGAAGAAAGGCTCCAAACTGTCAGGGTTCGT- GAACATCACATGTT
CTCGCTGTCTGATCCCCCC SEQ ID NO:72 Middle LPS-089
GGTGCGATCCTCAGGGTAATGGCCTGGCTGAATCAAGTAACAAGAATCT- TATAACC
ATTATCTAAGAAGATAGTAGGAGATAACAAGCGGTCTTGGGACAACA- AAATCAAGT
GCGCTTTGTGGGCAGATAGGATAACTAAAAAGAAAGCCACTGGTA- AAAGTCCCTTT
GAACTTGTCTATGGCATGGATTTGACATTACATGCCCATCTTA- AATTACTAGCTTAC
CAACTCCTTCAACATTTTTCTAGTGATAAAGGTGTTGTCC- AAAACATGGTTGATCAA
ATTGTGCAGTTGGATGAAATCCGCAGGAAAGATTTTG- ATAGTGCAAAAATCAGTCT
CCATTAAGAAAATCTTTGACAAATCTTCTCGGTCT-
AGATATTTACAGGTTGGAGATA TGGTTTTACTATGGATTCCACC SEQ ID NO:73 Late
LPS-090 GGTGCGATCCTGCAGGCTTAGATAGTTTCGGCGCTCCTCT- GAAAGAAGCACGAGT
AGGTGTCTCCACATTAGGTGGCCTGATCCCTTGCCTGCA- CTTGCAGCTTGTCTTA
CAACATCTCCTATGCTTTGATCCAGGCTTTTCACTGAC- ATAACTTCAGGGGCTTCCT
TCTCCCAGGGCCGTGCTGCCATCCAGCGTTCTAGC- CAGCTCCATCCCCAATTTGG
CTTGTTTGGGTCAATTTCCATCAGCATAGGATGA- GCTGCTCCTCGTGTGCTTTTCAA
TGACTGATGAGAATATGCGTTATGCCAATGC- CCTTTCTCGCTTCATGGCTGCTTCTT
GCTTGCTTTGCAAACTAGCCTCAATTTC- CTCTTTGGATTGCAACTGTCATCCAATCC
TTTGCTTCCATACTGGATCCAC SEQ ID NO:74 Late LPS-091
GGTGCGATCCCAAATGAACATTCAACATTCGATC- ATGTCAAGCGCTAAATGCCTTG
GCAGCTTAAAAGCTAGACTCCGCAAGTGACCC- TTCTGACTTAGTACACATATTAAG
CTCATCAAGGGTCCAATTCCATGAAAAGAA- ATTTTAAAACGGTTACATATTCACAAG
AACAGCACGAGATTTCCCAGATAGTCA- ACCACCAACTTGCCCTATCAGCCCAAATA
TTACTCATTCCATGTTAAAAATAGC- AAATTTCCAGATAGAATGTCGAAAGAGATCTT
CATGCACCATATATGGACTCTTAAAACCAGCCAAAATCTATACTGCCATGCTTGGAT CGCACC
SEQ ID NO:75 Late LPS-092
GGTGCGATCCTGGAGAGAGAAGCAAAAAGCCTACCATCTAAATCTACATCTAAAT
CAGATATCTTTACTGTGAAAGGAATTGAATGCTGCTTCAGATATCCTACAAGAATA
AGAAGAAAAGAATGATCAACTCCAAATCAGGCAGATGGCTCAGAATTTCCCGCAGC
TTCATTTTCGACGGCCTCCACAACACCAACCTCGGCAGGACGTATTACTCTGCCAT
GAAGTGTATAGCCAGGCTTCAAAACCACAGCCACACTGCCAGGCTGCTTACTAGCA
TCTTGAACTTGAGATACTGCCATGTTGCATATGAGGATCAAACTCTTCATTTATTGG ATCGCACC
SEQ ID NO:76 Late LPS-093
GGTGCGATCCCCAGAGGTTATTTGGGTTCAAAGTATTCTACACCAGTTGACATGT
GGTCATTTGCTTGCATAATTTGAACTGGCTACAGGTGATATGTTATTTGATCCTC
AGAGTGCAGAAGGTTATGACCGCGATGAGGACCACCTTGCCCTGATGATGGAGCT
TCTTGGAAAAATACCTCGTAAGATCGCCTTAGGTGGGAGCTATTCACGGGAACTTT
TTGACAGGCATGGGGATTTAAAGCACATTAGACGGCTTCGGTATTGGCCCTTGGAT CGCACC SEQ
ID NO:77 Late LPS-094
GGTGCGATCCTAAACTGTATGTCTCCACAATTGTCTTCAATATAGAAGCAGCTACG
CCCCTCCTAAGTCATCATAAGTTAAAAACTTCATCTTTCCAATACAATTAAACTATCT
AGCTTATCAGTTTGGAATAGAGATACAAAATTACAGATAGATTAGCGAAACTGTGCC
ACAAAACCTCTTCAAAATTAGAAGCATGATTGTCTACAACTCCACTTCAAAAAGGAG
CTGAACCAGTCCTTCGAAGGGTGTGCTTTGGTTGTGGTGGAGGTACAGAAGGCAG
CTAATTTCTCCAAGAACTGCTGTTTTTTTAGCCTCTCATTCTCCTCTTTAAGCTGCATC
ACTTCATTCTCTAGCTCATTTGTGTATGCCTGCTTTCTTGCCCTGGATCGCACC SEQ ID NO:78
Middle LPS-095 GGTGCGATCCGAGTGATGGCACAAAGAAA-
AGCAATGATAGAAAACAAAGAACAGGT AGCTCAGAAGGTTCAGCAACTAGAGAG-
TCAACTTCGAGTTAAGGAGGGCGGGAG CAATTGGCAGATTCTTCCAAATTTGTC-
AAGATCTCTTGGCATGAGATGACCTTATAG GATGTTAAGGAGCAAGAGGATTCT-
AGGAATAATGCCAAGGATAATAAGACTAAAAG
GATGCTTCAAGACCAGGTGGCAAGGAAGGCTTCTAATTCAAAGGGAGTTAGCAAC
GGCAACAGATGCAATTCTAGGATCGCACC SEQ ID NO:79 Middle LPS-096
GGTGCGATCCTAGAATTGCATCTGTTGCCGTTGCTACTCCCTTTGAATTAGAAGCC
TTCCTTGCCACCTGGTCTTGAAGCATCCTTTTAGTCTTATTATCCTTGGCATTATTC
CTAGAATCCTCTTGCTCCTTAACATCCTATAAGGTCATCTCATGCCAAGAGATCTTG
ACAAATTTGGAAGAATCTGCCAATTGCTCCCGCCCTCCTTAACTCGAAGTTGACTCT
CTAAGTTGCTGAACCTTCTGAGCTACCTGTTCTTTGTTTTCTATCATTGCTTTT- CTTT
GTGCCATCACTCGGATCGCACC SEQ ID NO:80 Middle LPZ-001
ATCTAGATCATCGATCTTGTCCAAATTTTAACTAGTGAATAGTTTAAAAAAA- AGCAA
CTAGCAGAAGAGAACCTAACCACTGACAAATTGCAAATACTCTAGAACA- CTATTCAT
CATTTTTTGCGATTCACGCTGGACCCACAAGAACCCCTTGAGCTGA- ACTTTCTTTTC
GTTCTCCCTCCTTTTGGATCGCACCATCTAGACCATCGATCTT- GTCCAAATTTTAAC
TAGTGAATAGTTTTAAAAAAAAGCAACTAGCAGAAGAGAA- CTAACCACTGACAAATT
GCAAATACTCTAGAACACTATTCATCATTTTTTGCGA- TTCACGCTGGACCACAAGAA
CTCTTGAGCTGAATTTCTTTTCGTCTCCTCCTTT- TGGATTGGACATCNAATCCTGCA
GCCGGGGATTCATATTCTTAACGGCGCNCGC- GNGGACTCCATNCCCCATATGATC
TTTTCATCCTGGCGCNTTTAACTCTGAAGG- GAAACCGGNTTNCCCTTATCCCTGGA
NATCCCTTCC SEQ ID NO:81 Middle LPZ-002
GTGGAGTGTAAAGGTCAACGTGCCATCCGGGTACAAACTATTGTA- GAAAAAATGGC
AAAGTTAGGTCTGAAAATATCCATTTGGCCTGCTCTAGTTGTA- CAGTACATGATTTT
GCACTCGCACAACAATGGACTATAATTATTTTCCTGGCAA- AAAAAAAAAA SEQ ID NO:82
Late LPZ-003
GGTGCGATCCAGGACATGAGGCCGAGTTTGCCATTGTGATATGATTGAGGAAGTC
CAGTCCTAAAATAGGTTTATCTTGATGTTTGACAAGAGATATAGAGGGGCATGATG
ATTCATTGATCTGTTTGCAGATCTGTAACTGCAACCATTCTAATGACATAATAGCGC
TATTGTTGGGTTCGTGTGATGACATAATAAATTGATTTAATTTAATAACATCTGTTA
ATGCAATGGCTGTAGCTGCATCATCACCGTATCCATCGAATGTTCCATTTTTCCAAA
TGTTTGTTTCCAAAACCAGAACACCAAAATGTCCCCTGCGTTTGTNTTGAAAAATAT
TGGGCCCNTACTATACTATAATNTTTNGGCATACTATACTATAATGTTTCTCCCATT- C
CCCCCAAATGANTCCTATACAATCCTGGCCGNCTTTACACTCCTGACNGGAAA- CCC
GGCTTNCCACTAATCCCTGGNCNANCCCTTC SEQ ID NO:83 Late LPZ-004
GGTGCGATCCGACTGTGATATGTGACTGGTGAACGAGAGATCCTTCTTATG- AATTA
ATCTGGTATCTTTATGCGAAAGCTTTTAGGGTTGCTACATGCTCTCCTC- TTTTGTAT
GAATTTCCATTCTAATATCAGTCTCTGTGAT SEQ ID NO:84 M,L LPZ-005
GGGGAGTGTCAAGGGATAAGTGGTAAGCCAGGTTTCCAGTCAGAAGT- GTAAAGGC
GGCCAGTGATGTAATAGATTCATATAGGGGAATGGAGTCACCGGGG- TGCGCCGTT
TTAGAATAGTGGATCCCCGGCTGCAGGATTTGATGGTGCGATCCT- GCCCCTGATAA
TTTGGTTGCAATGGAAAATGCAGTATTAGGTGCGAGATGTAAA- GCCCGCCCGGAG
CGGTGCATGAAGTACTGCAATATTTGTTGTAGTAAATGTGCT- GGTTGTGTTCCCAG
CGGTCACTATGGCAACAAGGACGAGTGCCCCTGCTACAGA- GATATGAAGTCCGCA
GCCGGCAAGCCCAAGTGTCCCTGATCTAGCACTCAGTCC- AGTCGCTCACTTCTT
TTATTCTTTTTTTTTATAAAAGTGACGAGGCCGTTTTTC- TTGTACTTGGTGGCCATAT
GTAGAGCGGTGGCTACTTCTCCTGTGTTAGGAAAT- GTTGCAGTACTAATAATAAGA
ACTTCTTTGGCAAAAAAAAAAAA SEQ ID NO:85 M,L LPZ-006
GGGTTTCCTTAAGAGTTAAAGGCGCATGATGTATAGAATCA- TATAGGGGATGGATT
CCCCCCGGGGGGCCTTTCAGAATAGGGATTCCCGGCTGC- AGGATTGATAGTGCGA
TCCAAGACACAGTGGAGTACCACAATGGGGATCTGGCC- AGTGCTTTGTGGCTATTC
ACTGCAGCTGTATTAAAACAGGAAGCCGCAAATGGC- CAGAAGGCCATTGAACTTGC
TGAGAGCAGACTATCTAAGGATGGCTGGCCTGAA- TATTATGATGGGAAGCTTGGAC
GATATATGGAAAGCAGTCTCGAAAGTGGCAAA- CCTGGTCAGTTGCTGGATATCTT
GTAGCCAAGATGATGCTTGAAGATCCATCCC- ATTTAGGTATGATAGCATTGGAAGA
GGACAAAAAGATGAAGCCGTCCCTCACTC- GATCAGCTTCTTGGATAATGTAAAATG
GGGAAATCCTAAACTTTCAGGCCACTC- TTGAATGTTTTGTCACTTCTGTATGACAAA
TGAGGCAATTCATAGTACATGTTG- TGCAAAAAAAAAAAA SEQ ID NO:86 M,L LPZ-007
GGTGCGATCCCAGAGAATATTAGTTCATGTGTTGCTCTCATTTTCTTCAATATGCAG
GGCAACCATTTGAATGAAATTATTCCTTTCGAATTTCAAAAACTTAATAGGCTAACTT
ATCTATCTGGAGCCGATTTTCATTGACGAGTAACCTGTAAGCTGGCCAGCAAAAGC
CAACAGATGTTCAGCTCGTTGGAACCAGTTGAAGATGTAATAGAGATGGTGAATA
ATCGCGGACGGCTCGGCCAATGGAATATTTGTTGCATCATCATCAAGGGGGTATGA
ATTCCAAAGAACTTGTTGATTGAAATTCCCAAGCAAAATTCTGTGAAATGAAAAATTT
ATTGAGACCATTGGGCAAAAAAAAAAAAA SEQ ID NO:87 Late LPZ-008
GGTGCGATCCAAAGAACACAAGATGGAGTTACCACAATGGAGGATCTTGGCCAG- T
GCTTTTGTGGCTATTCACTGCAGCCTGTATTAAAACAGGAAGGCCGCAAATGG- CCA
GAAGGGCCATTGAACTTGCTGAGAGCAGACTATCTAAGGATGGCTGGCCTG- AATAT
TATGATGGGAAGCTTGGACGATATATTGGAAAGCAGTCTCGAAAGTGGC- AAACCTG
GTCAGTTGCTGGAT SEQ ID NO:88 Late LPZ-009
GGTGCGATCTGTGTGGCTCTGAAACATCCCGGCTCCCCTCTGCACTATAATAATCC
CAAAATTAAGTGAACCCAACAGAATTTGCTCATATCTCTACAGTTATTGCAGACTGA
GCAAAACCCTCAAACTCATGTGACCTCTCAATAGGAGCCCACGCCCAAGATTTG- TC
CAGCATGTAACACACCTGATCGCCGCCACTGCAAGCACAACCGCTCACAAAT- ATCT
TGTCACACCACACTGTTGCGCAAGTTAACAATATTCATGTCTCCAGGAAA- GAAATGC
CACACTTCCCAACATTCTCTTTACTATTATAGAACTTCCTTGTTGCT- ATGGAAAAAAT
ACATTCCCAACGCAGAACCCCAACGGGGGTTCCCAATANCCCA- TTTCCCCCCTNTC
CAANCCNNTNTGAATGCNCCCCATNCCCTATTGNATNNTTT- AAATCCNGGCGCNTT
ANCTGGAAGGNAACCCGNTTCCCN SEQ ID NO:89 M,L LPZ-010
GTTTCCCAGTCAGGACGTGTAAAACGACGGCCAGGGATTGTAATACG- ATTCACTA
TAGGCGAATTGGAGGTCGATCCGTATAGGTAGTTGGATGATGAACG- GGCAAAGAA
GGCAAAGGAGTACAGTGATGGATCCTGTAATTCCTGTTTCAGAAA- ACAGAAAATCT
GCAATATAAGGATGGCTAAGCTTTTCAGCTATGAAAATATATG- GTGCAGTGGCACT
CATATCAGTTGCAGAGTTGTCAATATAACTTTTGTGAATAG- GAAAGTTGTCCTCTTT
TAGAGTGCAGAAATCCTGCAATATAAGGATGGCTAAGT- TTTTCAGCTATATGAAAAT
ATATGGTGCAGTGGCAAAAAAAAAAAA SEQ ID NO:90 All LPZ-011
GGTGCGATCCTACAGAGAGCAGCTTGACGAGGGCCAAAAG- GTTAAGGATGAAGAA
TGACCTCAGCTAGTAAGGTTTACAGAAGCAGCAGAGGCA- TCTTAACTGTTTTTATGT
TTTGGCAAAAGTTGTTGCGTCGGTTGTTTAATCCAG- GATTTCAGATGTATTTTGTAG A SEQ
ID NO:91 Late LPZ-012
ATTGTAATACGACTCACTATAGGGCGAATTGGAGGGTCCGATCCTGCGAGACCGA
GGGTTCATTTTCCTTTAGACAACGACGTTCAGTGGCGACCAGAGTTTCCCAATCAC
TTCAGCGATTCTATTCCTTCGTTGTAATAAAGCTTAAGGAATCCATGCAAATTCCT
GGAAGGTTTGAATATTTATATTTATTGGCAAAAAAAAAAAA SEQ ID NO:92 Late LPZ-013
AGGTGACCGTCAAAATGATTGCAGAGGACTTAGAGAGGGAAAAC- CGTTCCGATCT
GGTGAAGCAATTGGATGAAGCAGCTCTGGAATTGATTCCCGTT- CTGATGATATCG
TACGGCTAAGCTCAGCTCTTCAGGCAATTGGCAGAGAATACG- ATTCTTCAAATGAG
ATGACAGATTTTAAGAAACTTATAGATGAACATATTTCCA- AGCTTGAAGCGGATTCC
CCTACGGTCACCT SEQ ID NO:93 Late LPZ-015
AGGTGACCGTAAAATACTATGAGAAATGCTTTCATCAGGCACCGCTGGTAGGTT- TT
CTTCAAGCTTTTCATTAGGCAAAAGAGGCTCCGTGAGTTGATCGTAATCTCT- CCT
TGAATGGCCATATTGACCAGACACTCTGATTAGAAACTGGAATACAACTGC- ACATAT
AGTCATTCTTATATGATCATCCTTCTGCACTTCAGCATCCTGCGGCAA- CTCTCAT
CCCGCCATACTGCAGAAAAATTATTTGACTCTTGATCATGTTGTAGA- TGAATCTTCA
TGAATCTTCTCATCTTGCATTCTTGTCTTTATATCTTTAGGAAA- TTGCATCTGGTAAA
AGTATAAATGCATCTTCACTGGTTGCTTCAGTTTTGCATG- CTCCTGTCTTCTTGTT
TACATGTGATCTACCAAATCATCTAATGTATTCTCTCA- ATGTCTTGTGGACATCTCC
TTCATTCCGAGATTACCAATCATCTACCCGAATAA- ATGTGCCCCGTCAGCAATGCC
GTTTTGGTCC SEQ ID NO:94 Late LPZ-016
AGGTGACCGTAGTAGGCGTCCAGAGGCTGACAAAATCCCAGGCCTGTGCAAATC- T
GGAAGCCGCATGCAGGGCCGTGGCACCTACACTTGCGGCCTTAACAAAGTGGC- C
CGCGGCACCCACTTCTACCAGTGTGTTTATATTCTTGTGCAGCCAACACCAGA- GGT
TATGCAGGCGAATGTGCTGGCCAAGCGTTGTTTCGGCTGTCCGCAAACCCT- CTC
GAGTCTTACATGCCGCATATGAGTCTTGTGTATGGCGATTTGCCTGACGAC- GAGAA
AGAGAAGGCCAAGGTTAAGGCGCAGCTAAATTCGATGAACTTATCCGCA- ACACGG
ATTCCAAGTCTCCAGCTTGTGCTTGTACTCGACAGATCTGAAAATAAT- CCTCACTCA
TGCATAAGTGCAAAATGTGATCTTAACCTGCTCTGAAAATTACAT- AA SEQ ID NO:95 Late
LPZ-017 AGGTGACCGTCCACGAGAATTTGGCTTCA- AAACCCTAGGAGAGGGATATGAACTTG
CCAAGGCACAACTGACGCATGAACAAG- ACGTAAAATGACTCATTAGACACTGACAT
GATAATGAAAAACCTATGAATGATG- ATAGACTCAGCTACTTGATGACATCGCCCGC
CATTTGGACATCTTTATAAGGAG- TTTAAGCAAACCCTAGACCTACTGCCTAGTGACC
AACTTTTGCTTGACGACTCACTGAAATGACAATATTTGACCTTGACACTTCAAAATC
ACTTTGTAGGAACTCATTTGATCACTGGAGGACGGCTGGAAAGACTGACACTAACA
GGACTTTATATATGCACCTCGTCTATCCGAACTT SEQ ID NO:96 Late LPZ-018
AGGTGACCGTAAGCACAAGTCGTCAAAATTATCTCTATTCCGGCAGTAAAAACCTAT
AGCTAATGATGGATCAATAGCACTAAGTGGCAGCTGGCGTACATCACTGCAATGAT
AAGAACCAGTATCAACCCCCATATTATCAGGAGATATCTCCACCACCTGCTGCACT
ACATGTGGATCTAAGTACAGAGCCTGATCATCCTGAACACCAACAATATACGTTG
GCTCCAGGCTTTCCACCAGCAATACCAAGACTTTGGGGAAATGTGAACGTTTCACG
AAGTGATGGTACATACCTGGGTTGATCTCTCTACACCAAGAACAAGCGGCACCA
AAATCAGGATAGGCACTTGGTCTTCCCCTTCTCCATTGGACCACTCTGAACACA- GC
CTCGCAGCATCATCAATGCAGATAACTGGAGTCCCTCCACGGTCACCT SEQ ID NO:97
Middle LPZ-019 AGGTGACCGTGAATATGGTGGGTATTTGCAGGG-
CAAGATTCAGGATGCTGCTCCC GGAGCTTAAGTAAGGTCTTGGACCCTAATAAA-
TTCAGGGTATATGCATTATGTATAT GCTCTCATTTAGCTGCTCATCTGATTTCC-
ATTGGGTGAATCAGTTGTTTTGCAGTAC GTGGGGGTCTGTTATTTTGTGAGTTA-
TGGTGGAGTCATTTTGTTGTTGTTGTT TTTCTTATCTAGGGTTTAGGGTTTTGC-
CCTGTAATCGGTCTTCCCCTCTCTCCTGCG CTTGAATTTGACCTGAAACCTCTT-
GAAGTAGGCCCTGGTTTTCTGGGCTTTGACGA
AAACCATGGTTGTGGATCTCCTCTCTCCTGCTACGGTCACCT SEQ ID NO:98 Late
LPZ-020 AGGTGACCGTCCTACTTCACCGCAGTGACTTCCATCTGGTTTTAGGAAACTATCCC
TAAATCCTTCACTAGTTGACGAATTGATTGACTCAAATCAACTGTCGGTCAAACCCA
CTCTCTCTGAAAGTGAATTCTATGAGTCTATACCCAACCCAAATCAATAGGTTG- AGG
TAACAGTTGACCCGATTTCACCTTCAACAAATCATACCTTTCCCGAAGAGA- GTGAAC
ATGATTCAACACAAGTTCTTTTTGGTTCACCAGATTCAAATGAGCTTG- GGGGTAATC
CTCCTGTTCCATCAAGACAAGAAGAAAATCCTCCCACTCTCGTAA- CTCAAGGGTTAA
TCCTCCCATTTCTACGGTCACCT SEQ ID NO:99 Late LPZ-022
AGGTGACCGTCNCGGGATAGNTGGAGCCNAACAAAGTACNGAANAAA- NTGAANCG
CNCTGGGAAGCGNGCNGAAANNTGGNCANACNTGCCCTNCNACTCG- GTTACCCAG
CCNTTCTCTACCNANAATTATNACNNNANAGCNCCATGCTGGGTT- TGTNANAAAAN
AACNGCTNTTGATAAAATTACATAGANTNNNGAACACGTTAAG- AGGAATATGGTCC
ANATNCATTNTNAATNANNANTTAAAAACTNNNTATGTNCT- AGNGTCNCCT SEQ ID NO:100
Late LPZ-023
AGGTGACCGTACAGCACAGGTATACAAATCATAGAAATGGGCTTCTGTCCAACTGT
CAGCAGAAGCGATATGAAACCCAGAAGCATCAACTCTGCTTTCAATTTTTCAAGCG
CTTCATATAGAGCCTTTTTATTTCTCTGGAGAGCCAATTGCTAGCATAATGAATAC
CATGTTCAAGAAGTAAAGAGATGACCACAAATGCCAAACAAACAACTGCTACTGCC
CAAGTTAGGAGTTTGCTCTAGAGAACGGTCATTGCCACGGTCACCT SEQ ID NO:101 M,L
LPZ-024 AGGTGACCGTGGATATGGGAGCAGAGCCGTCCGCAGTGGATGCTGCA- ATTCAACT
TGAAGTGGCAGAAGCTGTGAAGACTCTCCAAATGGACAAGGCACGA- AGACAAAAC
CAAGACAAGGATGAGGGCAAGAGTGGCAACGCTGATTCAGATGAC- TTGAATGAAAT
GGAAGTCAAAGCTAAAGCAGCCGAACAACTGCTTGCTGTGCAT- GGGGCAGCATTA
CTACAGAATGCTCTGAAAGAAAATTGTCGAGTCATGAAATGC- GGGTTGGTTCAAA
TACAAGGGAGGAAGGTGAAGTTAGAAAGAACAGAAAGGGCA- TCAACGCAGACCCC
TCACTGATATCGGCAACACTACGGTCACCTAAGCCAATTC- TGCAAATTTCCATCACT
GGCGGGGCCCGCTCCAACTTCCTCTAAAAGGCCAATT- CCCCTATATGATTCTTATT
ACAATCCCTGGCCCTCCTTTTCCACTTCT SEQ ID NO:102 M,L LPZ-025
AGGTGACCGTAGCAGGAGAGAGGAGATCCACAACCAT- GGTTTTCGTCAAAGCCCA
GAAAACCAGGGCCTACTTCAAGAGGTTTCAGGTCAA- ATTCAAGCGCAGGAGAGAG
GGGAAGACCGATTACAGGGCAAGGATCCGCCTGAT- TAACCAAGATAAGAACAAGT
CAACACACCCTTGCCAAAAAAAAAAAAAAAA SEQ ID NO:103 Middle LPZ-026
AGGTGACCGTATGAGCAAGGAGGGAACAGTA- TGACAGGCAGTCAAAGCCCACGAG
GGGTGCCCCACTGCCTGCAGCAGCGCACTT- ACTTGGACTAACAAACTTGTATCGTG
ATTAAAACGATGAACATCGTATTGTGGA- GTGGAGCCACTCGTGACCTGATTCTGTC
CTAAGTACTTGGTCCTGGAATACAAT- ATTGCACGGTCACCT SEQ ID NO:104 All
LPZ-028 AGGTGACCGTCAAAGTACAATGGAGTCATATATCCACTTGAATTGAAACCTCTAATT
TAAAAGTTCTCAAAAAATATTTTATTTACAAAACAGGGAAAATAAAAAATGACTCTAT
CAACTATACAATCCTAACATCCATCTCCCGACAGACCTCCAGTATATGTACAAGGC
GCTGAAAGAAGGCTGATTATTTTCTATTCCAGCTCGCATAACGTGGTCTTCTGAG
GCTTTGCCTATTCCTTTCTTTAAAATCTTTCGCACGAAAGATTGGCATTGACCTTCG
GCTAAATCTCAGACTCCAGGGAACCTTGGACTCCCTTTAAAACCTAGAGCTACTTTT
TACGAACCCCTGCTTCTCTTGAACACTTAGGGAACTTATACTTACAAAACTTCGGG- A
ACTCCACCCCCTAGCTTTGCAGGACTCCAGCAGATTCCCCAAACTGCCAGAAG- GC
TATTTCCATGCACTGTTAGGGGTGAATTCCTACTATCAAAACCCCCAAAACA- TCATA SEQ ID
NO:105 Late LPZ-029 AGGTGACCGTATGGGAACAAGTATG-
GGAACAAGAACGTATTACATAAAAGATGGA GATGCAACACAGCATAAATTGATG-
CTAAGTTGTTACAATGATGCATACAGCTTAAC
CAAGCTTGGAATGACATCATTAAGTGCGGTCACAGCCTCTGCATAGTATTTCTCT
GCCTTGGGTGTATCCTTGCTCCTTGCAGCGTAGTCCAGGTTGTCAAGGGTTGTCAA
AAAGCTTGGTGGTGAAGGTTTTGAGGGGCTTCTTCTGGTCCTTGGGCTTTGAGGA
GATAACGGTGTTTGAAGTCCTTAGCGAAAGTAAGAAACCTTTGGAACCGAAGTCCG
TTCTTGACGTTACCGCACGCCTTCCTTATCTATCACTTTTTCACCTCCAGAAATTGC
TTCCCGAATCCCTTGCTCTCCCACCCCCTGTCCCCC SEQ ID NO:106 Late LPZ-030
AGGTGACCGTAGTGTTGCCGATATCAGTGAGGGGTCTGCGTTGATGCCCTTTC- TG
TTCTTCTACTTCACCCTCCTCTCTTGTATTTGAACCAACCCGCATTTCATGA- CTCGA
CAAATTTTCTTTCAGAGCATTCTGTAGTAATGCTGCCCCATGCACAGCA- AGCAGTTG
TTCGGCTGCTTTAGCTTTGACTTCCATTTCATTCAAGTCATCTGAA- TCAGCGTTGCC
ACTCTTGCCCTCATCCTTGTCTTGGTTTTGTCTTCCGTGCCTT- GTCCATTTGGAGAG
TCTTCACAGCTTCTGCCACTTCAATTGAATTGCAGCATCC- ACTTGCGGAACGGTCT
GCTCCCCATATCACGGCACCTT SEQ ID NO:107 Late LPZ-031
AGGTGACCGTAGTGTTGCCGATATCAGTGAGGGGTCTGCGTTGATG- CCCTTTCTG
TTCTTCTACTTCACCCTCCTCTCTTGTATTTGAACCAACCCGCAT- TTCATGACTCGA
CAAATTTTCTTTCAGAGCATTCTGTAGTAATGCTGCCCCATG- CACAGCAAGCAGTTG
TTCGGCTGCTTTAGCTTTGACTTCCATTTCATTCTAAGT- CATCTGAATCAGTGTTGCC
ACTCTTGCCCTCATCCTTGTCTTGGTTTTGTCTTC- GTGCCTTGTCCATTTGGAGAGT
CTTCACAGCTTCTGCCACTTCAATTTGAATTG- CAGCATCCACTGCGGACGGCTCTG
CTCCCATATCCACGGTCACCT SEQ ID NO:108 Late LPZ-032
AGGTGACCGTCGTGAAATAGCGAGAACGGCGTGGAACAT- CGCAACGGCGGGGAG
GCTGGCGGACGTTGCACGTTTCTGGAAGGTATGCGGCTC- TCTCCTCCGCCTCAGT
TTCCATGAAGAGGTCCTCCCTGGTGAATCATACGATTG-
CGATTGATCGAGTACTT GCTGTATGGCTCGGCATCGGCATTGTGGAGACATTCT-
TTCCTATTCCTCGCAGCAT CTCTCCGATGGTTGCTCTCTCCGGAGCTCCATGTT-
ATCCCCGGCACTGAGACAGTC GCTGCCGAATCGCAAGAGCTTCTTTGTTTTTTG-
CAGGCTTCTCCAAACATAATGCCT CCGGGCCCCTCAACCGAATTCTGCCAAATC- CACCCC SEQ
ID NO:109 E,L LPZ-033 AGGTGACCGTGGACGACAGTGAGTG-
CAGTCATCATGCTCTCCAGTGGACTTTAAG CAATCTGCATCTTTATGGAAGTGA-
TGTATCTCTTGTGGTTTTTCATGCTCAACCATT
GGCAGTCTTCAACAGTGCTGCAACAATGGGCATAACGTCTCCCGAATTAATTGAAA
CTATTGTGAATCAACAGATAGGTTTCTGGTCACATCTAGCAATACAAACACAAATAA
CTGTGGAACAGAGCCACAAAACTATGCTTCAGAGCATCTAATTACACATATCTCTC
TAAAACCCTTGCATAAAAAATAAACTGAATCTCGACCTTAGCACTATTGCCACCATC
ATCTCAAGCAAACATTCTCTAGAATACCATCTTCACAATGCACTAAAGTTACATAAG
CACTGAACTTAAAACATTTCTGTGACGAATGAAGGACCAATTCATCATACTCAGCCT
TTGCATCCAATCTGTTGAATGTGCTGAAAAATGCCCAATAAACCTCCATCCAACACT
GTCTTCCTCTCTGAGGTGCACACTGATTTCTGCTGCTGAACCAGTCGGGATTCC- CT
GCTCAACGTCCC SEQ ID NO:110 Middle LPZ-034
AGGTGCCCGTGGAACTACTGTTAAATCTGGAATCCCTTGTCTAGCTGTTAAAAACTC
GACAAGTGCATGTTGGTATAGTAGGGTAACAGAAGGGTTCTTACCCAGATTTAC
CCCTTTGGCGGAGATATTTAAAAAAAAAGAATTGTCATTATGGTAAATAGGTGTGAC
AGGTTATCAATAGAATAACTGACGAGAGTAAACTGATAATTATTAAGGTTAAAGTGT
TCGTAAAGGAGACTTGGACTCTAGGTTGGATGCCTACACTTAGAGCCGTCCCGCA
CTTGGACGGTCACCT SEQ ID NO:111 Middle LPZ-035
AGGTGACCGTCCAGTGCGGGAACGGCTCTAAGTGTAGGCATCCACCTAGAGTCCA
AGTCTCCTTTACGAACACTTTAACCTTAATAATTATCAGTTTACTCTCGTCAGTTATT
CTATTGATAACCTGTCACACCTATTTACCATAATGACAATTCTTTTTTTTTAAATATCT
CCGCCAAAGGGGTAAATCTGGGTAAGAACCCTTCTGTTAACCCTACTAATACCA- AC
ATGCACTTGTCGAGTTTTTACAGCTAGACAAGGGATTCCAGATTTAACAGTA- GTTCC
ACGGTCACCT SEQ ID NO:112 Late LPZ-037
AGGTGACCGTATGGGAACAAGAACGTTATTACATAAAAGATGGAGATGCAACACAG
CATAAATTGATGCTAAGTTTGTTACAATGATGCATACAGCTTAACCAAGCTTGGAAA
TGACATCATTAAGTGCGGTCACAGCCTCTGCATAGTATTTCTCTGCCTTGGGTGTA
TCCTTGCTCCTTGCAGCGTAGTCCAAGTTGTCAAGGGTGTCAAAAAACTTGGTGGT
GAAGGTTTTGAAGGGCTTCTTCTGGTCCTTGGGCTTTGAAGAAATAACGGTGTTGA
AGTCCTTACCAAAGGTTAATAAACCTTTGGAGCCGAAGTCGTTCTGGACGTACG- GC
CACCCCTTCCTTATCTATCAGCTTTTTCACCTCCAAGAATTTGCTTCCCCGA- ATTCC
TTGCTCTCCCAGCCGCCTGGTCCCCCGAAAAGGGCTGAATATAAAACCG- TCCTCA
ACGGCATTCCATTCCTCCCTCGTCTGAAACACTTCCCCGCTGCCCCCG- AGGTGAA
GGGCCATCAACTTGATGAACGGCTTTTGCAAGGCTCTGACCCCGGCC- CCGTCACT
AACCAATTCTGCAATC SEQ ID NO:113 Middle LPZ-038
AGGTGACCGTGGGGAACAACTACATGACAAATCATTTCTTTGTGGTGGATGTACTG
GACACCAAATAAGTGTTGAGAGTCCACTGGCTCTGTACGCGTGGCAGAATCACAAC
GGACTTGAGAAAGTTGAAGATGGAATTTGTATCGCTAGATGGCCAGACCATGTT- GC
TTCAAGGGATGCACTCGTAACCCCCACAGTCTGTCTCTACCCACTAGATGGA- GGCT
GACATGAGACATGGAGACATTAATTGGGTTGTGGAGTTAAAGATCTCTCA- CGTTCG
GGGAAAATCCAAGCCATCATACTTATATATCCGTCCCGTGCATGTAAC- CTCCTCCA
CTCTGTCCCTTAGGCCCGTTGTTGCCT SEQ ID NO:114 E,L LPZ-039
AGGTGACCGTATGAGCAAGGAAANNACCGCACTGGCTCCCAGCAGCA- TGAACANC
CAGGTCCCAACCATANACCNCNTGGAGAANGTGATCAAGATATTAG- CGACAGTGTN
ATTGTACNTCTCNCCAAACACATTATACACGATAAGAGAGCNTA- AACTACTCTATTC
CTTTGACGNAGTGACTACNTGAGTANAAGCGATCATTATCT- TGCNAACTTTGCATG
AAAACAACAAACCCACNTCCAGTTTCTCTATANTCTGGC- CCCACNATGAATAANANT
CCTGCCATAATAATGANTCTTTGTCCCCANAGANAA- ATTNNATAAGACAGGAGCCC
ACTGTTGCTTGCATGACTACCANTCACTTTAAGG- CGTTGCGAATCCCGGTCCTAAC
CATCTCCATACCATNGGCANNCTTTACTTTCC- AACTGCCCAAGACTGTGAACAGGG
CGGTTCNNACCCTATAANTTTTAGCCTCTN- NTCGAANCNCTTNTTTTCGTTCCCCGG
AAANCCGNTTCCCACCCTTTGGAACCT- TTTTTTTTTGCCGGGCCCCAGGCNAATTC
TNCAATTCCCCNCTGGGGGG SEQ ID NO:115 Late LPZ-040
AGGTGACCGTGGCGGAGGTTAGGGAAGTTTGACTT- CTCATTTTCTCACGCACTCCT
CTCCCTCGTAACCTCGGTCGAGTCGATGGCGGC- TTTTTAGTCGAGTGTGCTAACG
CACCCTCCGGGCCTCAAAATTTCCAGCTACTC- GTATTTGATCAATGCTGAAATCGC
GTAATCACGTAGATAATAAAGCGTAATGAA- TTCTATAATGAAGCATGTTTCTCTATA
GTTCATGTTGCCGAGAAGGAATAATGA- AAATGAAGCCTTATATATTATCTGGGGCTC
AAGGAGATGTTATCTTTTCTCTTC- CTTGGTTAGAGACCGTCACCTTCACTTTGAATT
GGATAAAGCTTCATTTGTTTAAGACCTCCCACCCGTAAATACATACGGTAGCCTTCT
TATGTTAGAAACATACGTCACCTACGCAGAATTGTTAGAATGAAATGA SEQ ID NO:116 Late
LPZ-041 AGGTGACCGTGGAACAAGATGATTAGTTCTCATGCGGGCCAGGATGATTA- GTTCTC
CTATGGCAACTGTTGGACAGGATGATTCGTTCTCCTGTGGACAGGATG- ATTAGTTC
TCCTATCGAGGCATCCTACCCAAGCAGTTTGGGACTCATGGGAAGT- ACCTCTCATC
TGATCAATGAGTAGGAAATGGGGTTAGGGACCATTAAGTAGTAT- TATCGATGGATG
CATTGTTGTATCTATTGTACTCCCTATGCTAGAATGAACTCC- ATTGATCTGGGATCA
ATGAATACTGTTTCTGGGAATCATTGAAAATTTGTATGA- ACACACTCTGAACACTGA
ATTTCCGGTTCATTGGAAGAGATGGTTTTAAACACT- CTCCTCATCTCATTTCTTCCC
CTTCCTTATTCCAACCAAATTTGGGCCACCCTG- CCAGGAAATTCATTTGATGGTTGG
AAAATACCACGGGCCCTAACCAATTCTGCA- A SEQ ID NO:117 Late LPZ-042
AGGTGACCGTNCATCTCTACCATNATNCC- TCCCTCCCGNCTGTATCANCNGGCNTN
NANGTCNTTNNCTANNNNAAGNTAATC- CTATCCCNTTANAGTGACGGTCTCTAN
NCCTAGAAGAGAANCCATAACATCTCC- TTGAGCNACACATGGGATATACCGCCANC
TTATNTAATACTTTCNCNGCACGGT- AACNGACCANAANCATTCTTCACTATAGAATT
CATGTCGCTTCATTATCTACCTCATTNCNCCANATCCCCCTTNATCTCATNNATTTA
CTAGAAANTTCTGAAGNTCCNNAAGGGTTCGTTTTGCACCNCCCCAANTAAAAAAN
CCCTNCCGNTTACNTCGAACGAAGGTTTCAAANGAACAGNAATTCCTTTACAAAAA
TCAANAATTTTAACTTCCCNAATCCGGCCCCCCNGTNCCCGAAACCCNATTTCTAC
GATTGCATCACCCCGGGGGNCCNCTCAANCCNNCTCTAAAGGNCCATNCCCNT
NNNTGATCCTCTNCCATCCAANGGCNCCTTTCCACTTTTATTGGAAAACCCCCNTT
CCCCNTTTTACCCTTNNAAGGCCCCTCCC SEQ ID NO:118 Late LPZ-043
AGGTGACCGTGGAACTACTGTTAAATCTGGAATCCCTTGTCTAGCTGTAAAAACTC
GACAAGTGCATGTTGGTATTAGTAGGGTTAACAGAAGGGTTCTTACCCAGATTTAC
CCCTTTGGCGGAGATATTTAAAAAAAAAGAATTGTCATTATGGTAAATAGGTGTGA- C
AGGTTATCAATAGAATAACTGACGAGAGTAAACTGATAATTATTAAGGTTAAA- GTGT
TCGTAAAGGANACTTGGACTCTAGGTTGGATGCCTACACTTAGAGCCCGT- TCCCGC
ACTGGACGGTCACCT SEQ ID NO:119 Late LPZ-045
AGGTGACCGTGGGGGATGGGGCCGTGGGGAAGACTGTATGCTCATCTCCTACAC
AAGCAACACGTTTCCAACGGATTACGTGCCGACTGTTTTTGACAATTTTAGTGCAAA
TGTGGTGTTGATGGCAATACAGTAAACCTTGGCTTGTGGGACACTGCAGGGCAA
GAAGATTACAACAGACTGAGGCCATTGAGTTATAGAGGTGCAGATGCTTTTCTGCT
TGCCTTTTCTCTGATCAGCAAGGCTAGTTATGAAAATATATCAAAGAAGTGGAT- TCC
AGAACTAGACATTATGCACCAAATGTGCCAATCATTCTTGTGGGAACTAAA- TTAGA
TTTGCGTGATGACAAGCAGTTCTTTGCTGATCATCCTGGAGCAGCCCCT- ATAACAA
CAGCTCAAGGTGAAGAGTGAAGAAGCAGATTGGAGCAGCAGCATATA- TTGAGTG
CAGTTCCAAAACCCAGCAGAATGTCAAGGCTGTTTTTGATGCTGCTG- CAATTAAAGTGG
TTCTTCAGCCACCAAAGCAGAAAAAGCGGAGAAAAAAGCAGA- AAAATTGTTCTATTC
TCTAAGAAAAATGTGGATGTTCTGAACGCNCTTCACTGA- CAATAANGNTGACGTNG
GAATATCTTCCTCC SEQ ID NO:120 Late LPZ-047
AGGTGACCGTAAGCACAAGTCGTCAAAATATCTCTATTCCGGCAGTAAAAACC- TAT
AGCTAATGATGGATCAATACCACTAAGTGGCAGCTGGCGTACATCTCTGCA- ATGAT
AAGAACCAGTATCAGTCCCCATATAATCAGGAGATATCTCCAGCACCTG- CTGCACT
ACATGTGGATCTTAGTACAGAGCCTGATCATCCTGAACACCAACAAT- ATACGTTGAA
GCTCCGGGCTTTCCACCAGCAATACCAAGACTTTGGGGAAATGT- GAACGTTTCACG
AAGTGATGGTACATACCTTGGGTTGATCTTCTCTACACCAAG- AACAAGCGGCACCA
AAATCAGGATAGGCACTTGGTCTTCCCCTTCTCCATTGGA- CCACTCTGAACACAAG
CCTCGCAGCATCATCAATGCAGATAACTGGGCGCCCTC- CACGGTCACTT SEQ ID NO:121
Late LPZ-049
AGGTGACCGTGCCATAGCGCATGGCGTGTAACTGGATGAGACCGCATGGCTCAAA
TCTGCTAGGAATCAACATGAAATCAGCTCCAGCTGTTATCATATGAGCAAGTGGCA
CGTTAAACTTTGCTACTCCCCTGACGTTGTCTGGATATTTCTCTTCAAGCTCTTCAA
GCTGCTTCTCCAAGTACTTTTTACCGGTGCCTAGGATAATTAACTGCACGTTTCAT
CTGCAATTAGAGGGACAGCTTCAGCAAGAATATCTGGACCTTTCTGCTCTTCAAGT
CTTCCAATAAATCCTATAACAGGAATATCTGGATCCACGGTCACCT SEQ ID NO:122 Early
LPZ-051 ATGTGACCGTCAAAAGGGCATATAAATCGGGGAGCTCAATGGCAA- GAATGTACGAT
TTCTGGCCTCAAGTCGCCCTGAATTTGGTCAACAACATCTTGA- TAGAGCGAGAGGA
CGCTCCCAATTAAGATCTGGAAACTGTCGAGAGTGATTGAG- GTCATTTTTAATCTAA
ACTGAATTGTGGGGACAATTTTTCAATTCAGATCCTTC- TAGCAAAGCAAAGCAAACC
TTAACAGTATTGTATCCATGAGAATGGATTCTGCA- CAGGTCAGGCTCCACGGTCAC CT SEQ
ID NO:123 All LPZ-053
AGGTGACCGTGGAGAAGAGAACGCTTTGCCGACTCTCTGGGATGCCCTTCCCTCC
ATAGCCGTCGTGGGAGGACAGAGCTCCGGGAAATCCTCTGTGCTGGAGAGCATCG
TTGGAAGGGATTTTTTACCGCGTGGATCAGGTATTGTTACTAGACGGCCGCTTGTC
CTTCAACTTCACAAGACTGATGAAGGCAGCAGGGATTACGCCGAATTCCTTCACCA
ACCCAGAAAGAAATACACCGACTTTGCACTGGTAAGGAAGGAAATTGCGGATGA- GA
CTGATCGAATTACAGGGCGTTCCAAGCAAGTCTCAAGTGTCCCAATCACCTT- AGT
ATTTATTCACCCAATGTTTGTAAATTTGACTCTAATTGATCTCCCTGGGTT- GACAAAA
GTGGCTATTGACGGTCACCT SEQ ID NO:124 Middle LPZ-054
AGGTGACCGTGCAATATTGTATCCAGGACCAAGTACTAGGACAGAATCAGG- TTA
CGAGTGGCTCCACTCCACAATACGATGTTCATCGTTTGATCACAATACAGG- TTTGT
TAGTCCAAGTAGGTGCGCTGCTGCAGACAGTGGGGCAGCCCTCGTGGGC- TTGGA
CTGCCTGTCATACTGTTCTCTCCTTGCTTCAGGCTCTACTGCTGTTGCT- GCTGCTG
ATACGGTCACCT SEQ ID NO:125 Middle LPZ-055
AGGTGACCGTACATACAAGGTCTTATCACCAGCAGCAAGAATAATCAGTTGGCCAT
CTTCTGCAGGCTTCTTGCTGCCTGAGACAGGAGCCTCAAGAAATTTTCCCCCCTTT
TCAATGATTGCCTCATTGATCTTTGTTGAAGTGATAGTATCAACTGTTGACATG- TCA
ATGTATCCTTTTCCTGTACACATTTGCTCTAGGACACCATCCGAGAGGGCA- GCAGG
AGGATCAGACAGGATGGCTATGGTATAGTTGCACTTCTTTACAACTTCG- GCAGGAG
TGCTTCCTATGGAAGCACCTTGCTGAACAAGTTCTTCACACCTAGAC- ATTGTCCTAT
TCCACACGGTCACCT SEQ ID NO:126 Late LPZ-056
GGTGACCGTACATACAAGGTCTTATCACCAGCAGCAAGAATAATCAGTTGGCC- ATC
TTCTGCAGGCTTCTGGCTGCCTGAGACAGGAGCCTCATGAAATCTTCCCCC- CTTTT
CAATGATTGCCTCATTGATCTTTGTTGAAATGATAATATCAACTGTTGA- CATGTCAAT
GTATCCTTTGTCCTGTACACATTTGCTCTAGGACACCATCCGAGA- GGGCAGCAGGA
GGATCAGACAGGATGGCTATGGTATAGTCGCACTTCTTTACAA- CTTCGGCAGGAGT
GCTTCCTATGGAAGCACCTTGCTGAACAAAGTTCTTCACAC- CTAGACATTTGTCCTA
TTCCGCACGGTCACCT SEQ ID NO:127 Late LPZ-057
AGGTGACCGTGGAGGGGCTCCAGTTATCTGCATTGATGATGCTGCGAGGC- TGTGT
TCAGAGTGGTCCAATGGAGAAGGGGAAGACCAAGTGCCTATCCTGATTT- TGGTGC
CGCTTGTTCTTGGTGTAGAGAAGATCAACCCAAGGTATGTACCATCAC- TCGTGAA
ACGTTCACATTTCCCCAAAGTCTTGGTATTGCTGGTGGAAAGCCTGG- AGCTTCAAC
GTATATTGTTGGTGTTCAGGATGATCAGGCTCTGTACTTAGATCC- ACATGTAGTGC
AGCAGGTGGTGGAGATATCTCCTGATAATATGGGGGTTGATAC- TGGTTCTTATCAT
TGCAGTGATGTTCGCCACTGCCACTTAATGCTATTGATCCA- TCATTAGCTATAGGTT
TTTACTGCCCGGAATAGAAATAATTTTGACAACTTGTG- CTTACGGCACCT SEQ ID NO:128
Late LPZ-058
AGGTGACCGTGGAGGGGCTCCAGTTATCTGCATTGATGATGCTGCGAGGCTGTGT
TCAGAGTGGTCCAATGGAGAAGGGGAAGACCAAGTGCCTATCCTGATTTTGGTGC
CGCTGTTCTTGGTGTAGAGAAGATCAACCCAAGGTATGTACCATCACTTCGTGAA
ACGTTCACATTTCCCCAAAGTCTTGGTATTGCTGGTGGAAAGCCTGGAGCTTCAAC
GTATATTGTTGGTGTTCAGGATGATCAGGCTCTGTACTTAGATCCACATGTAGTGC
AGCAGGTTGGTGGAGATATCTCCTGATAATATGGGGGTTGATACTGGTTCTTATCAT
TGCAGTGATGTACCCACTGCCACTTAGTGCTATTGATCCATCATTTAGCTATAGGTTT
TGCAGTGATGTACCCACTGCCACTTAGTGCTATTGATCCATCATTAGCTATAGGTTT
TACTGCCGGAATAGAAAAATTTTGACAACTTGTGCTTACGGTCCCT SEQ ID NO:129 Late
LPZ-059 AGGTGACCGTGCTAGGACACACAATTTCTCAGCAAGGAT- TACAGGTGGATCCTAAC
AATATTGCTATAATTCAAAAGGTTCCACCTCCTTAAA- AGGTAAGAGATGTTTGGAGT
TTTCTAGGCTTGGCAGGATATTATAGAAGATTCA- TCAAAGATTTCATTAAGCTAGCC
TCGCCATTGTCTAGCCTCTTAGGGAAAGATG- TTGAGTTTCAATGGACTGATGACTG
CCAAGGGGCTCTGGATGAGTTGAGAGATA- AGCTGGTATCCGCCCCGATCTTGAGA
GGTCTAAACTGGGCCCTACCTTTCCACA- TCCACATTGATGCCTCGAACAAAGCCAT
AGGGGCAGCCTTAGGACAAGTTGAAG- AGAAAATACCATATGCCATATACTTGTCA
GCAAAAATCTGTCTAAGGCAGAACT- GAACTATACGGTCACT SEQ ID NO:130 Late
LPZ-060 AGGTGACCGTCATATTCCCCTCTATAGCAGCACTAACAATCCATTTTCTGAGTGCAT
CAGAAAATCAACACACGGTAAATGTCTTGAGACTAACGAGAAATTAATAATCACGTT
GTACAAAGAACAGTATGTCCCGTCACGTCACGAGTGCCCTGAGAGATCATCCAACT
TTCTCTGAACCCTCGTGTTACACGCACGCAAAATCAAGGATCAGTTGTAGTTATTGC
TGGCGTGACAGACGTGACACCTACTGTTCCGCTACAAACGATATAATTGAATCCAT
GATCGGATTATGTATTATGATCTTAGCGCAGTGGTTATGAAATTATGATGAATTTGC
TTATGATTTTCTCAGCGTTTGTGGAAGAATCTCGCTATTGAAAACTTCCCCGTATAT
TTCCAAACTTATTATCATCCCACGGTCCCT SEQ ID NO:131 Late LPZ-061
AGGTGACCGTACAGCATTTATTGATGTTCTATTTTGTTGTTTGCAAGTTTTTC- CGA
CGCTGTGAGGCACGGAAAACGAGATAAGTTGTAAAAGTTTGCTCGCTGATT- TGAGG
CACGGAAAACGAGATAAGTTGTAAAATTTTGCTCGCTGATTTTTTGCTG- AATATTTC
TCTCACTATAAAAAGCATTTTCCAGAAATAAGAAGGAGCTTTCGAA- CTGGTTTTCCC
CAAGAGTTGTAGGGGGTTTTTCCACGGTCACCT SEQ ID NO:132 Late LPZ-062
AGGTGACCGTATTTATGGTCGCAGGCACAAATTCTGCTA- CTGTAGAAGGGTTCTTA
CCAACTTTAGGTAGAAGGCGAGGAGGGCTTTATTAGT- ACAGTTCTGTGTAATCTTA
ATGATATTTTTTGCACTATTATTTTATGGTAAAAG- GATTGATTTGTCTTTTGCAAAGG
CCTTAGGATTGTTTATTTACCTTTGGGCTAA- GGGAGGAGGTAAATTTTTCACATTGG
GAAAAAAAATGCCTCGGTCGTTGTCACG- GTCACCT SEQ ID NO:133 Late LPZ-063
AGGTGACCGTGCCAGTATGACAG- ATGGAACCATGCAGCTAGCCACCAAATTGTAAA
CATCAAATTTTGTCTCAATATAAGTTGCAAATTCTTAATTAATTATGATCACCATTTC
AACGGTCACCT SEQ ID NO:134 Middle LPZ-065
AGGTGACCGTGAATAGAAGCGAACACATCCTTGTTGCTGAATCTAACGACCAATCG
GTATTTGGGTGTGTTGTACTTGTTCTTATCTTGGTTAATCAGGCGGATCCTTGCCCT
GTAATCGGTCTTCCCCTCTCTCCTGCGCTGAATTTGACCTGAAACCTCTTGAAGTA
GGCCCTGGTTTTCTGGGCTTTGACGAAAACCATGGTTGTGGATCTCCTCTCTCCTG
CTACGGTCACCT SEQ ID NO:135 Middle LPZ-066
AGGTGACCGTGGTAGAGGAGGCAGGCACTCATCTAACAGTCGAAAGCCCTTTACA
AAGGGGAATGGTACCAGCATAGAGAAGAAACACAGACGGTTTGAAGAGGATGATG
GATCTGCCATAGATGAACGATCAAATAAGGTTCAAAAGCTGGAAAATGATGGTGAA
TTCCATGCATCCCACTTGGCTCTGTCCCTCAAGTTGAATATACCTGGACGAGAGGT
ATTGCATTTCCCAACGGTCACCT SEQ ID NO:136 Middle LPZ-067
AGGTGACCGTACTGATAATAGAAGAGGCAGGGAAAGAGAAATCAATGATAATAGAA
GAGGCAGGGAAAGGGAGATCAATGGCATCATGCTACTTCTGTAGCTGTTACCT
TAGTGATGTAATCTTCCATGGCAGACTCGGGGGTTTTATCTTTAAGTTGAATTTCCA
TGCATCCCCTTGGGCTCTGTCCTCCAGTTGAATATCCTGGAACAAGAGGTTTTGCT
TTCCACGGTCCCCT SEQ ID NO:137 Late LPZ-069
AGGTGACCGTGAGAAGGCAACTTTATCCCCTGCTAAACCAAGTCCAGAAATGAGGA
AAATATGTGAAAACTGAATTGCTATATATGATGCCTAGTCTTGGCCTCTCAATTACA
AGTTCAACGTCTTCAAATGATTGAAATATGGACCTTCTTAACCGTTCTGGAAATCTA
TCAATCTTCAAAATTTTGAAACTTTGCCTCGATCTTGGAGTGATCAGACTTGATTTCT
AATCCTAGAAATACCCTATCACTGGCTACCTGGTCTGTACGGTCACCT SEQ ID NO:138 Late
LPZ-070 GGTGACCGTGGGATAGGCAGAAGCAAGAAACACAGAAGTTCTC- CGGGAATGTAA
GCGCTGACAGTGGGGGAGAAAGTAGTGAACAAGGACATGGTCG- GTATGAAATACA
TGGCAGGCGATGGATTTCAAGGGATTAAGCATCTCAATGGAT- ATTTACTATTGGAC
TGTAGTAACTTTCGCCATCGCTTTTTGAACACATCTGTGG- CTTAACTGTCATCTGTA
ATGGTAAGCGAACCAGGTTTTGTTCTGAACCACTTGT- ATGTACGGTCACCT SEQ ID NO:139
Late LPZ-071
AGGTGACCGTGGTGGAGCGATTAGTGATTGTGATAAAGGGAGCATCAATATCTATG
TAGACGCCGTATAAAGGTGGAAAAGGTATGTTTTGCAGGTATTTCTTTGTAAATGGT
TTATAATGGGTTAAGCTCGGATATATGAGGTTTATATATAAGTCCTGTTAGTGTCAG
TCTTACCAGCCTTCCTCCAGTGATCAAATGTGCTCTAACAAAGTGATTTTGAAGTGT
CAAGGTCAAATTATGTCATTTCAGTGAGTCTTCAAACAAAATTTGGTCACTAGGCAT
TAGGTCTAAGGGTTTGCTTGAACTCCCTCTAGAGTTGTCCAAATGGGCGGGCTATG
TCATCATTTAAGCTGAATCTATCATCCAATCAATAAGGTTTTTCATTATCATGTCAG- T
GTCTAAATGAGTCATTTTACCGTCTTGTTCACGGCTTCACTTGTGCCTTTGGC- AAAT
TCAATTCCCTCCTCCAAGGGTTTGAACCAATTCTCTTGGACGGCCCCTAA- ACCAA
ATCTGCAAAATCCAC SEQ ID NO:140 Late LPZ-072
AGGTGACCGTGGTGGAGCGATTAGTGATTGTGATAAAGGGAGCATCAATATCTATG
TAGACGCCGTATAAAGGTGGAAAAGGTATGTTTTGCAGGTATTTCTTTGTAAATGGT
TTATAATGGGTTAAGCTCGGATATATGAGGTTTATATATAAGTCCTGTTAGTGT- CAG
TCTTTCCAGCCTTCCTCCAGTGATCAAATGTGCTCTTACAAAGTGATTTTG- AAGTGT
CAAGGTCAAATTTTGTCATTTCAGTGAGTCTTCAAGCAAAATTTGGTC- ACTAGGCAT
TAGGTCTAAGGTTTGCTTTAACTCCTTCTAAAAGTTGTCCAAATG- GCGGGCTATGTC
ATCATTTACGTCTTGTTCAGCTCAGTGTGCCTGGCAATTCAT- TCCTCTCTAAGGTT
TGAACCATTCTCTTGACGGCACTAAGCCAATCCACACTGG- GGCCGTCTATTGAATC
AACCCGGACACTGGGTTACAGGCAAC SEQ ID NO:141 Late LPZ-073
AGGTGACCGTCCAAGAAGAAATTGGCTTCAAAACCCTAGGAG- AGGGAAATGAACTT
GCCAAGGCACAACTGAAGCATGAACAAGACGTAAAATGAC- TCATTAGACACTGACA
TGATAATGAAAAACCTATGAATGATGATAGACTCAGCT- AAATGATGACATAGCCCGC
CATTTGGACAAATTTTAGAAGGAGTTAAAGCAAAC-
CTTAGACTTAATGCTTAGTGAC CAAATTTTGTTTGAAGACTCACTGAAATGACA-
AAATTTGACCTTGACACTTCAAAATC ACTTTGTAAGAGCACATTTGATCACTGG-
AGGAAGGCTGGAAAGACTGACACTAACA GGACTTATATATAAACCTCATATATC-
CGAGCTTAACCCATTATAAACCATTTACAAAG
AAATACCTGCAAAACATACCTTTTCCACCTTTATACGGCGTCTACATAGATATTGAT
GCTCCCTTTATCACAATCACTAATCGCTCCACCACGGTCACCT SEQ ID NO:142 Middle
LPZ-074 AGGTGACCGTGATAGACCCAAGAAAAATAGATCCAACCCTCAGAGGGACAA- AGA
CTTATAAAGACTAGAAGAGTGAATCAACCTATTCTATTTAGAATATATATT- TTTGGGG
TGCTTGCTTATCGTTTTGGGGGTTAATGTATGTCGTACTACGGTCTT- ATGCCCTAAT
TTGCCCATTGAAATCAACTAAATTGACAGTAACCGACTAAAAGT- TGGTCCACACTAA
GATATCGATGACCAACGATCATAAAGGTGTCCATGATCCTA- ATAGTATATGTGTCAA
TTAATGTAACTTTGGTGCTACAACATAAAACCATTCGT- GGGGATCCTCCTTTTTATG
CGGTCACCT SEQ ID NO:143 Middle LPZ-075
AGGTGACCGTGGGACCGACCTTGACTACAGGCCAAAATTTTGACTGTTGAC- CAGC
GTTCACTTCTGTATTTTTGGTTGGTATGAGCAACATTGACTTGCTGGAAA- TTGACCA
GGTTTGACTGGTATTTGGACTTGGATTTTGGCACAGATTTCTAGACA- ATTTGTATTT
GTAAACCTTACAGAAGAATAATTTATCGAAGAAGAAAAATGCTA- GGTTTCCCCTCAA
GTTTGGGTTTCCCAAGGGAAAAATTGTTGTCCCAATGGTTG- AATTTTCCAAAGGTCT
CCTAACCCGACAATACCTCCTAAGAATTCCTTAATTTA- ACCTTTCTTGTTTTCACGGT CACCT
SEQ ID NO:144 Middle LPZ-076
AGGTGACCGTGAAGGAGCAGCAACAATTTGATTTTGTTTGGGTAGATCGGG- GATTT
TCTCGTGGAACATACCTGATTGAGTATAAACTAAGTCAAGGTACTGTGC- TTGAGAAA
TTACTTGCTCCTCAGTAACTACTCTGGCCTTAGCTACATCCTCAGT- GATCTTGGGTA
GTAAAGATTTTACAAACCATTCAGCTAAGATCTGATCCGGGAT- ATAAACTTTCACTA
AACGTCGTCGACGTCTCCATTCATGGATATGATCTGAAAT- GTAAGTGGACGTTGAC
TGCTTTAACGAAGTTAATAATTCTGTGCCATTTTCATA- TCTGACGGTCACCT SEQ ID
NO:145 Late LPZ-077
AGGTGACCGTACCTAATGGGAAGACACTTCAAGGTAAAAACAAATCATGATAGTCT
TAAATACCTTTTAGAACAAAGATTATATTCAGAACAACTTGCTGGAAGTGTACCAAG
TATGACTGGTATTGAGACTTAGATCTTCGCACAGATTTCAAGACAATTTGTTGTTGT
AAGACTCACTCACGAAAAGTGATGTGGATATGAAGAACTTCCCTGTCGCCTCTTGG
TTAGGAGTCTCCCACTCATAGGAATTGTGTAACTTATAACTTGGTCCACTAAAGAAG
TTAGGTACAGTGTGTTCCTTTACCAGGTTCCCTGTTGTAACTTACAAATCTACGGCT ACCT SEQ
ID NO:146 Late LPZ-078
AGGTGACCGTCACTGGAGGTTTGAGATGCTTGATCGGTACTGAAATGAGACATGAT
CAGAATAGGACCTTGTTGAGGCCGTGTCTCACCCCCCATCCACAATCTTTTGTAAT
TTTGAGTTTCGTTTAGAACATACTTGTAGGATAAAACTTACCTTACTCATGGATCAT
GGCTGTATATGTTTATCGACCAGAGACAGATATGCCGAATGAAAGCGAGTCTAGTA
TTCTAATGCAATATATTGGTAGTATGGGACATAGTACTGAACACTTGTATAGTACGG TCACCT
SEQ ID NO:147 Late LPZ-079
AGGTGACCGTGGTCTCAGTTATGCCATATGTCCGCCCCTCCATATGATGCTCCGCC
TCTATGGGGGTCTTTGCGATGTTGATATCTAGTAGTACTTCTTGTCCTATTGCAGCA
ACCTGTACTGGTGTTGGTGTTGGTATGGGTCTCCTACGCGATGGAGATATGAGAC
ACCCATAGGTCGAACAGGTCTAATATCTGGAATCCAACGCTATTGTTGTAGAAG
ACGTTGCTCCCGTCCTTTAGCTTTGGCTGGTCACTATCCTTACGCTCCACGTACGG TCACCT SEQ
ID NO:148 Middle LPZ-080
AGGTGACCGTTGGGAAATGCAATACCTCTCGTCCAGGTATATTCAACTTGAGGGAC
AGAGCCAAGTGGGATGCATGGAATTCACTTAAAGATAAAACCCCCGAGTCTGCCAT
GGAAGATTACATCACTAAGGTTAAACAGCTACAAGAAGTAGCATGATGCCATTGAT
CTCCCTTTCCCTGCCTCTTCTATTATCAGTACGGTCACCT SEQ ID NO:149 Late LPZ-081
AGGTGACCGTCAAGGCAAAGTGTCATGCCACTCATTGGAATTAGTTAATATAG- CTA
ATTTGAGATATTACAGTCAACTGTGGGTATATGTATGTGAGATCAAGGTGC- AGTTTA
GATATTATCAGTGGTGCAGTTTAGATATTATCAGTGTTTGTGAATCTG- CATACTGCT
TTTGGTTGGTTCTAACTACGGTCACCT SEQ ID NO:150 Middle LPZ-082
AGGTGACCGTAGACATATATCATGGAAAACCCAAGTAACATACA- AACACAAAACACA
TGGAACTTCATAAAACCTCCACTCGTCATAAGCTTTATTGC- TATGTTATTGTGGTG
TTGCATCGTACTTAGTGGAGGTTATTGTTATGTTATGTG- TTCTATTTTCCTCCCGAA
CGCCCTTCGGAATTGAGCTAACCGTGGTTAACAACA- TGTGGGCTTTTTTTCTCGAC
AGTATATATATAATAAATCTTTATTTTTTTAAAA- ACTAATGCTATTGCATTTATATACT
GGAAAAAATGATTTTTCTGTATTATCGAA- AATAATAATTTAGTTTCTTGATAATCACT
TGGAATTAAGAAATTACAAACCCTA- ACAACATCAAGAAATTTTAAAACACATAAGCTA
GAAATTTTAAAACACATAAGCGTGACAACAAGAAGATCAAATCTAATACTTGCTTGG
GCCGGAGATTATGGATTCATGAAGCGATTTGACAGCGTCCATTGATCTTCCTCTCC
CACGGTCACCT SEQ ID NO:151 Late LPZ-083
GGGGGTAGGGGTGTTTATACTGAGCATACTTCGAAAGTGGTTCACCACCACCATG
TGACTAATTGTTCCTGACTTTGGTAGACCTATAATAAATTCCATAGAAACCTCCGTC
CATATTGATGCCGGAATGGGCAACGGTTGTAATGTGCCTGGTACTTTGACGGTCAC CT SEQ ID
NO:152 Middle LPZ-084
AGGTGACCGTTGGGAAATGCAATACCTCTCGTCCAGGTATATTCAACTTGAGGGAC
AGAGCCAAGTGGGATGCATGGAATTCACTTAAAGATAAAACCCCCGAGTCTGCCAT
GGAAGATTACATCACTAAGGTTAAACAGCTACAAGAAGTAGCATGATGCCTAGACA
AATAGCTTTGCTCAACACATCCTGATAGTGTACACTAAATCGCACAACTTTACTACT
ACAAAGAAAGATCGTTGACACCTTGACAAATAGCTTGCTCAACACATCCCAACAAT
TTGGATTGCGAATACCGACTCCAATTTGTACTTGATCCATATGTCGTTGCGATGTAC
TAGTTCCTCTATACATATGTTTCTGCAAGAATCGGAGTTGGACCTCTTCTTCCCTGT
TATCAGCACGGTCACT SEQ ID NO:153 Early LPZ-085
AGGTGACCGTGGATAAGAGAACGCTTTGCCGACTCTCTGGGATGCCCTTCCCTCC
ATAGCCGTCGTGGGAGGACAGAGCTCCGGGAAATCCTCTGTGCTGGAGAGCATCG
TTGGAAGGGATTTTTTACCGCGTGGATCAGGTATTGTTACTAGACGGCCGCTTGTC
CTTCAACTTCACAAGACTGATGAAGGCAGCAGGGATTACGCCGAATCCTTCACCA
ACCCAGAAAGACATACACCGACTTTGCACTGGTAAGGAACGAAATTGCGGATGAGA
CTGATCGAATTACATGGCGTGCCAAGCANAGTCTCAAGTGTCCCAATTCACCTAA
TATTTATTCACCCAATGTTGTTAATTTGACTCTAATTGATCTCCTGGGTTGACAAA- AT
TGCTATTGACGGTCACT SEQ ID NO:154 Middle LPZ-086
AGGTGACCGTTGGGAAATGCAATACCTCTCGTCCAGGTATATTCAACTGAGGGAC
AGAGCCAAGTGGGATGCATGGAATTCACTTAAAGATAAAACCCCCGAGTCTGCCAT
GGAAGATTACATCACTAAGGTTAAACAGCTACAAGAAGTAGCATGATGCCATTG- AT
CTCCCTTTCCCTGCCTCTTCTATTATCATTGATCTCTCTTTCCCTGCCTCTT- CTATTA
TCAGTACGGTCACCT SEQ ID NO:155 All LPZ-089
AGGTGACCGTACATACAAGTGCTCAGTACAATGTCATATACTACCAATACATTTGAT
TAGAATACGAGACTCGCTTTCATTCGGCATATCTGTCTCTGGATGATAAACATATAA
AGCCTTGATCCATGAGTAAGGTAAGTTTGAAGCTACAAGTATTTTCTAAACGAA- GTT
CAAAATTACATAAGATTGTGGCTGGGGCGTGAGAAACGGCCTCAACAATGT- CCTGT
TCTGATCATGTATCATTTCAGTACCGATCATGCCTATCATACCCGCCTG- GTGACGG TCACCT
SEQ ID NO:156 Middle LPZ-090
AGGTGACCGTACTGATAATAGAAGAGGCAGGGAAAGGGAGATCAATGGCATCATG
CTACTTCTTGTAGCTGTTTAACCTTAGTGATGTAATCTTCCATGGCAGACTCGGGG
GTTTTATCTTTAAGTGAATTGCCATGCATCCCACTTGGCTCTGTCCCTCAAGTTGAA
TATACCTGGACGAGAGGTATTGCATTTCCCAACGGTCACCT SEQ ID NO:157 Late
LPZ-091 AGGTGACCGTATAGTGTCAAGCTTTTCTGGATTGGATAATGGACGG- CGGCTTGCG
CATACATCTACACATTCTGTAACAAGTACACTCTACTGCAACAGC- AGACCCAATTTC
ACCTCTTCAGTCAGCCAGAGATCTCGATGGATTTGGGTTGAG- GAGGTTGGGGTTC
GCCTGCTTCGGCACGGTCACCT SEQ ID NO:158 Early LPZ-092
AGGTGACCGTGCTAAGTAATTATCATCTGTACCTGTGCTTGCTGC- AGGAAGTAAAC
CAACCCGACTAGTCTTTTTAATAATACAGGGAGCCTTGCCACC- AATTTCCTCTTGAA
GCACCCATATTGGACGGGTTTGTGTCATCCTCTGTATTAT- CCTTTTTCATCCCAAGC
AGGCTGTCTGTTTTTGTAGTAGAAGGATCACAACACA- GATCAGGCCCTCCATAGTA
CAAAGAAGAACCGAGGAAAGTATCATTAACGTTCT- GACTCCTGCCATGAAGGCTTC
CACTATGACCTTGACCCTTTTGTGAATTACTGC- CATTTAGACCTTGACTGGCTCTTG
CAACCAAATGCCCCAGAATGGAACTTCTTT- GTGCTCCAGTTCCATTGTGGTTAGTT
GAATCCCTACCACGGTCACT SEQ ID NO:159 Late LPZ-093
AGGTGACCGTGCAATATTGTATTCCAGGACCAAGTACT- TAGGACAGAATCAGGTCA
CGAGTGGCTCCACTCCACAATACGATGTTCATCGTT- TTAATCACAATACAAGTTTGT
TAGTCCAAGTAAGTGCGCTGCTGCAGACAGTGG- GGCACCCCCCGTGGGCTTTGAC
TGCCTGTCATACTGTTCCCTCCTTGCTCCTGC- TCTTGCTCTCGCTGGGCTGTGGTG
AGTTACTAACCTGGTTCGACCCACAAGGGC- TTCTCACTAGGGCGTTAGGCTGCATG
GATCTGCCAGATATTGTGGTTGCAAGGG- ACAGAGGCATGAGACACAGGCCTTTGC
TTTGCAGAAACTGCATTGCTGACCCCA- TGTTCATCCATCAGTTTTGCTACCTCTC
CTTCTGTTATGGACGGTCACCT SEQ ID NO:160 Late LPZ-094
AGGTGACCGTATCCGCAGCAGCAACAGCAGTAGA- GCCTGAAGCAGGGGACCTAAT
TACAGTCAAAAGTCCAGGGCTACCAATGCCTGC- TAACAGCGCACTTACTGGACTA
ACAAACTTGTATTGTGATTAAGACGATGAACA- TCGTATGTGGAGTGGAAGCCACT
CGTGACCTGATTCTGTCATAAGTACTTGGTC- CTGGAATACAATATTGCACGGTCAC CT SEQ
ID NO:161 Late LPZ-095
AGGTGACCGTATCCGCAGCAGCAACAGCAGTAGAGCCTGAAGCAGGGGACCTA- AT
TACAGTCAAAAGTCCAGGGCTACCAATGCCTGCTAACAGCGCACTTACTTGG- AACT
AACAAAATTTTTATTGTTAATTAAAAACGAATAACATCGTTTTTGTGGGA- GTGGAACC
ACTCGTGAACTGAATCCTGTCCTAAGTTCTGGGTCCTGGGAATAAC- ATATTGCACG
GGTCACCTT SEQ ID NO:162 Middle LPZ-096
AGGTGACCGTTACAGCTAGGGAAGACTTTAAAAGTTTGTAAAACTAAGCATAGCTC
TAAACACTGAAGTTAAAAGACATGATTGGAATGTGCAAGTGGTTCAGTATCCAAAT- A
TTGAAGGTTGCAGAATATGGAGCTACTGTGCAAACGAGTAACTTTATCTATAT- TTTC
ACAAGATCATACAATGGGAAACGTTGAGATAACAACTGCATCGGTGAACC- AGAATA
GTTATAAAAGTTCTTGCAAGTAAAGGGATGAATAATTGCATGGTTGGA- ATTAAGAAT
GACCATGTAGAGCTGCTATACAGATTCTCCAAGGTTTTATATTTG- AGGAGTGCGCG
CTATTGATGTTGTGCAAAAATTTCAGAAATTAAGTTCTGCGGC- ATTTATCAAGGTG
TTTGAGCCATTTAAATAGCAAGTTTTTGTTCTCCAAGTACT- TTCAGGAAAGCAGAT
AGCTCTAGTTATAATGCTCCAGTGACAAACACATCTAGT- TGGGGCAGTGAATGACG
CTTTTGTCATTCTCTTTTGGTTTCAGGCACGGTCACC- T SEQ ID NO:163 Early
LPZ-099 AGGTGACCGTGGACAAACTCTAGAACAG- GCATAGCTTTCATGTTCAGTTGTTTTTAA
AGAGCAGTCCTCGCAGCAGATCGTG- CAGCTTCCTGCTTCACTTCCGTTGATTTTCC
TGATCTGAAATACCCGTAAACTT- GCTGAAGAACCCAAATACTTAATAGCGTCTCTAA ACAAAA
SEQ ID NO:164 Late LPZ-100
AGGTGACCGTGCCTGAAACCAAAAGAGAATGACAAAAGCGTCAT- TCACTGCCCCAA
CTAATGTGTTTGTCACTGGAGCATTATAACTAGAGCTATCTA- CAAGCCAAAACAGTG
TTTGGGAGAGATTCCATAACGTCATTGCCTCTGCTACAC- ATCATTCATTGGTTCCAA
TAATGAAGCCACGTGCTAAGGACATTGAGAGAATCT- TATAAAACAAGAAATATAGTA
AATTGGGAAATGCATTTTATCGTCTAACCTGCT- TTCCTGAAAGTACTTGGAGAAACA
AAAACTTGCTATTAAATGGCTCAAACAACC- TTGATAAATGCCGCAGAACTTAATTTC
TGAAATTTTTGCAAACATCAATAGCGC- GCACTCTTCAAATATAAAACCTTGGAGAAG
TCTGTATAGCAGCTCACATGGTCA- TTCTTAATTCACACCATGCAATTATTCATCCC
TACTTGCAAGAACTTTATAACTATTCTGGTCACCGATGCAGTTGTTATCTCAACGT
TTCCCATTGTATGATCTTTGAAAATATAGATAAAGTTACTCGTTTGCACAGTAGCTC
CATATTCTGCAACCTTCAATTTGGATACTGAACCACTTGCACATTCCAATCATGTC
TTTTAACTTCAGTGTTTAAGAGTATGCTTAGTTTTACAAACTTTTAAAGTCTTCCCTA
GCTGTAACGGTCAC SEQ ID NO:165 Middle LPZ-101
AGGTGACCGTAAAATACCATGAGAAATGCTTTCATCAGGCACCGCTGGTAGGTTT
CTTAAGCTTTTCATTAGGCAAAAGAGGCTCCGTGAGTTGATCGTTAATTCTCTCCTT
GAATGCCATATTGACCAGACACTCTGATTAGAAACTGGAATACAACTGCACATATAG
TCATTCTATATGATTCATCCTTCTGCACTTCAGCATCCTGCGGCAACTCTTCATCCC
GCCATACTGAGAAAAATATTTGACTCTTGATCATGTGTAGATGAATCTTCATGAAT
CTTCTCATCTTCATTCTTGTCTTTATATCTTTAGGAAGTGCATCTGGTAAAAGTATAA
ATGCATCTTCACGGGTGCTTCAGTTTTTGCATGCTCCCGGTCTTCTTGTTTAGCAT
GTGGATCTAGGAAATCACTAAATGTAGTTCTCTCAATTGGTCTGGTGGAAATTC- TCC
TCAATTCGAGAATTACGAATCATCATACCTGAGTAATATATGTTGCCCTGT- ACATGC
ATATGCTGGTTTTTGGCTCCACCATCTCCAAAGGGCTCAAAAACTATG- CGACCCC
TGGTTGCCGTAGTGGAAGGTTATACATTGCGTTCCCAGTAGCCACGG- TCAC SEQ ID NO:166
Middle LPZ-102 AGGTGACCGTGGAGGGGCTCCACT-
TATATGCATAGATGATGCTGCGAGGCTGTGT TCATCTGGTCCAATGGAGAAGGG-
GAAGACCAAGTGCCTATCCTGATTTTGGTGCC
GCTTGTTCTGGTGTACAGAATATCAACCCAGGGTATGTACCATCACTCGTGAGAC
GTTCACATTTCCCCACTTCTTGGTGGAGCTGGTGGAAAGCCTGGAACTTCATCAAT
CTATCGTTGGTGTGAGGATGATCAGGCTCTGTACTTATATCCACATGTAGTGCAGC
AGGTGGTGGAGATGTCTCTGATAAGTTGGGGGTTGATACTGGTTCGTATCATTTGC
AGTGATGTCCCCCGCTGCCCTTAATTGCTATTGATCCATCATTAACTATAGGTTTT
TACTCGCCCGGAATAAGACAATCTTTTGACACTTGTTGCTTGGGTCAC SEQ ID NO:167
Early LPZ-103 AGGTGACCGTGGCGCCTGACCTGTGCAGAATCCATTCTCATGGA-
TACAATACTGTT AAGTTTGCTTTGCTTTGCTTGAAGGATCTGAATTGAAAAATT-
GTCCCCACAATTCTG TTTCGTTAAAAATGACCTCAATCACTCTCGACAGTTTCC-
AGATCTTGATTGGGAGCG TCCTCTCCTCTCTCAAGATGTTGTTGACCAAATTCA-
GGGCGACTTGTGGCCAGAAA TCGTACATTCTGCCATCTACCTGTTATTGAGCTC-
CCCGATTTATATGCGCTTTTGAC GGTCAC SEQ ID NO:168 Middle LPZ-106
AGGTGACCGTCAATACCATTAAACTGGGGATTCGTCTCAACAAGTCAACAT- GCTAA
CCTCACAGCTCCAATCAAACAACGTCCGTCGAAGGGCGCTCACACTCAT- CCAAATT
ACTTCCCTCTGCAAGACTCACAAAATCAGATTCTTCATGAATTGCTC- AAACGAGGCT
GTTATGGATGATGCAGCTGATTACTCAAGTGACAGCACTCTGAA- TCCCCGTCCCAT
ATATAGCGACGCGGCGTTTCAGCCGTGACTGGTCGCAACAGC- CTCAGTGGGACACAA
AAGGCCAGAAGCCCCCCAAGGTTCTCACGGTCAG SEQ ID NO:169 E,L LPZ-107
AGGTGACCGTGTCGATGTTGTTAGATGTGATTAGGG- TTTTATTTCTTGATACAGATG
CACTGTTTCTCTGTTTATTCTTTTATTTCTTCA- ATGTATGTTGTCAAATTATACTTAGT
CA1GATCTCCTTTTATCGTTCGTCAAAA- AAAAAAAAAAAAAAAAAAAAAAAAAAAAAG
TTTAACAATTAAAAGGGGAAATTA- GGCCATATCAGCTTGTCGTATGGACCCACATG
ACTGTAGGTCAC SEQ ID NO:170 Middle LPZ-108
AGGTGACCGTATGCAGAGTCAAGGTTTAGTTCCTTCA- GAGCCTGCCCGAGTAGCA
CTGAGGCAGCTCAAGCCATTTCACGTAGGAAGCCCA- CAACAAAATAGAAATCAGAG
TGAGTCTTTGATCGAGTAACCCATAAGTTCTTAG- CTCCCGTTCCATCTTAACATAAG
CATTTTTCTTCGTCTTCTCGCAGCCGT SEQ ID NO:171 Late LPZ-109
ATTGCAGAGGACTTAGAGAGGGAAAACCGTTCCG- ATCTGGTGAAGCAATTGGATG
AGCGCTCTGGAATTGATCCCGTTTCTGATGATA- TCGTACGGCTAAGCTCAGCTCT
TCAGGCATTGGCAGACAATACGATTCTTCAAA- TGAGATGACAGATTTTAAGAAACTT
ATAGGATGACATATTTCCTAGCTTGAAGC- GGATCCCCCTACGGTCAC SEQ ID NO:172 All
LPZ-110 AGGTGACCGTCCGATAAAGGATGAGAATATAGGTAGATCAACCCAAAAACACTCTC
AGAAAACGATTAAAGCCTAACCCCAAGATCGTTGAGTAAATTTAACCCGGTAACCTC
CACATAAAATATACTTAGCAACAATAAACTCAACAACTAAACTATCCCTTTAAAATTA
AATTATCCTATTTATTTAAAAAAACAAATCCTTTATATACTAAGGTCCCCTGCACAT
CTATTACTAAGGTAAAGGAAGGGAATTATATGCTATCATTGTAAACTTTGACTTCCG
TATTTATGATCAGACCATGAGTTTGATAATTAATTTTACGCTCTTTACTCCCCATTC- A
AGGCACGTGCCTGGTGATATATGAACGCCAAATTATT SEQ ID NO:173 Late LPZ-111
AGGTGACCGTAGAATACAATCTATGTATCTTAAATGCTAACAAAGA- GAATTTGTTGTC
TAGCTTGTAAATATACAAAAGAAACTCTCACAAGGAGTGAGA- AGCACTAAGGCCCT
TGGAAAGAATACGTTTCTATTCAGCGGAGTGTATTTTGAG- CTACGGCTTGGCACAA
CTCATCCTATAAAACAAGACTCTGTGAGAGGGCAGAGA- CCTTGATCCTGGGCGTG
GCAAGCCGGGTGCCTATTGCGGTAAAATCGAGAAGGG- GGACCCTGGAAAAGAGAG
GCTGAAATTTGTTTCATTCTGCAACTGAAACCTAAC- CGGAGGCCGAATCTGATCATT
TCTAAGACCTTTGGGGTCCTGGGCATCCCATTA- AAAGAACGCTGCTAACTCTCCCC
TCCACAAAGGGCCAATGCGCTCAGGTCGGGC- TTCTCATCTTCACATTTCTTGCCGA
AATCTATCTGAATTTGTTGTATTGAATAA- CACTGCCTCCTACACGGTCAC SEQ ID NO:174
Late LPZ-112 AGGTGACCGTGGGCGCCGTGGCTCAAAAGGCCCTCGCAGACGCCCGCTCCATCA
AGCTCATGGGCCCCCTCCACCCTCGGGGGGCAAGCCGGGAACGTTGCTGTCAGA
CGAGGCGAGGACCTGGAACTGCCGTTGAAGGAACGGTTCTATATTCAGCCCCTCT
CGGCGGACCAGGCGCTGCGAGAGCCAAGGAATCCGCGGAAGCAAATCCTGGAGG
TGAAAAAGCTGATAGATAAAAGGCGTGGCCGTACGTCCAGAACGACCTCCGCTCC
AAGGCTTCTTACCTTCGCTACGACTCAACACCGTTATCTCCTCAAAGCCCAAGGAA
CAGAAAAAACCCCTCAAAACCTCACCCCAAAGCTTTTTTGACACCCTTGACAAACCT
GGACTACGCTGCAAGGAGCCAAGGATACCCCAAGGGCAGAAAAAATACTTTGCAG
AAGCTGGTGAACCGCCCTTAATGATGTTCATTCCAAGCTTGGTTAAGCTGTATTGC
ACTCATTGTTAACCACACTTAACGCCAATCCAATCTATGCTGTGTTGCATCTCCACT
TCTTAGTTAATAACGTCTGTGTCCCAAACTCTGTGCCACACACGGTCAC SEQ ID NO:175 All
LPZ-114 AGGTGACCGTACAATACAAATAGGTAGTTTATCACATTGTAG- CTTATAGAATGTACA
ATTGAAATCAAATAAATTCAACCAAACTCAAATAATATG- ATCATGTGCTCCTCACCTT
CTCAGCAAACTCGTAGAGCAGAAAAAAGGATTATG- TTAAATCACAGTTCACACATTA
GGGTAAATCCCACTAAATGACCTCTCTTCATT- ATCCAAGTATCTGACACCAACATAT
TTCAAACAAATAGTGCAAAAAGGAATGGT- GAAGTAAAATAGTCAAAACTAAAAAATA
AGCTTAAAATTTCTCACATGTTTGAA- TATGTGCACCACAAATTTTGTTAGTGTCATCA
AAATGCATGTAATCAACTTGCCGTGTATATAATTTCACACAATATCCGTAAAATTTTG
CAATTCCTTATGAGCATTTCATGTCTAGAGATTGCAATGACTTGGCTACAAACATGT
TTCTCTACACAAGATCACAATATTTAGTCAGGACACGAATTGCAATGGGGATTCTCA
CAAGCATCACAAGTCATCTCCCATGTACTAATAAAAAATTGTTTAAAT SEQ ID NO:176
Middle LPZ-115 AGGTGACCGTATAGTGCATATTCAGATTGCAATTACAGACGTA-
TTAGAACCAGATTT TCGCTTCGATACAGCTCATCGAGAGCAACAGAGATCCAGA-
TCAAAAACCAGACACA GTTTAAGAACATCGAAATACCAAGCCCAGGGACAGTTA-
CCAGCATATAGCTCTACC ACCAACAGATTATTACAGAACCAAAACATAAGACCA-
CTTGCAGACAAAAATAAACCC TAACGCAGAACGTGGCAACTATCTCCTCCAGCT-
ACCACCATCGGAACCACCACCAC CATAGCGAGAACCCCACCACCACCATAGCCG-
CCACCGCCACCACCATAACCACCA CCACCACCACCACTGTACCGCCACTACCGC-
CATAACCACGGTCAC SEQ ID NO:177 E,L LPZ-116
AGGTGACCGTCCTTGGAGATACCAGCTTCAAAACCTCCAGTGGTGGAGTCGATGA
CAATACTGCACAGTCAGCCTGAGATGTTCCAGTAATCATGTTCTTGATAAAATCACG
ATGGCCGGGGCATCAATCACAGTGCAGTAGTATTTAGTTGTCTCAAACTTCCAGAG
TGCAATATCATTGTGATACCACGGTCAC SEQ ID NO:178 E,M LPZ-117
AGGTGACCGTATAGTAGGAACTTTAGGTGCTTTTGGTGGCACTCTCCAATTTTCATG
TCCTTACATACCCCACTACGGAGAAGGGTAGCCCAAGATTTGAACCCAAGACTTCC
GGTTCGTGAGACTTCATTTCCACGGTCAC SEQ ID NO:179 All LPZ-118
AGGTGACCGTAAGATCAAGAGCACAGAAAGCAGCCATAGCCCCGCCCATTGAATG
CCCATAACAATAATCTGTAACCCATCTCTCTGTTTCTGAGCTTTCTGAACTGCTTCT
ACAACAGTGGTCGTAAGGTTGTGTTGTGATAAGCAGAGTAAAATCCATAATGTACC
ATTGCACCAGCATATTAGGATAGTTGAGATCAAGTGTCTTACAGAATAAATCCT- CCA
CCCAATTCTGTAGCTCCTTTCTTGAGTACCCCTGAATGCAATTACAATTGC- ATTGAT
ATCTTCTGCCACACCACAAAAGCCTGAAGGCAGTGTTGTACATCAACT- ATAAGCTC
ACCACCTGAAAACCCCAGTCAAACCATTGCACCTAGAACAAGTCCA- AGACATTAGA
GCACTCAAATCATCCATATAGACCGCAGAAGCATATTGCACAAG- TATCTCAGCAAGT
GTTCGATTATAGACATGGCCA1GGTCAC SEQ ID NO:180 Middle LPZ-119
AGGTGACCGTGGGAGGGGAGATTTTTGATTTATATTTCCAA- TATAAAAGAAAATCTA
NGTTGTAAGGACATGGCAAGAGCTCTTATTTCCGGGGT- TTTAGCCGTGGCCCGGA
GCGGATGAAAGCAAATGTAAGTCACTCCGTGCTTCTC- GGCATTTGGACGCTTCTA
CTCTACCGCACTACAGACGGGATTGAACCTCGCATC- TCTGAGTGTTTGGTCGTTTA
CATGGCGGACTTGTTCCGCACCTCTGCGGACGTC- AAATGCCGCGACGATAATCCC
TTTGAGAACAGCGATACGGCAGAAAGATCGCCG- TTGACGAAGCGAGAAAACTATTG
AGACTTGCAGATGTGGAGCTGAAGAAGAGCT- TGAGTCGACGGTCAC SEQ ID NO:181
Middle LPZ-120
AGGTGACCGTCCGTTCGGGGTGTATTTGTCGAACACGTAGGATGGTGCTACGTTGA
AACCACCGTTACCTTCTTCGATATGTTATAGTTCGAGTTCATACGGAGGGAATACC
GTTTGTAGTGTTATTCAGCACAACCCCGTCCTGATTAAACACCCCCGCAACCAAGG
ACGTATTCGACGTTCGGTATTGTTTGACACACTCAAGTTATAACCCTGAATAGGCG
CTACCCGAAGTAAGCATTGTACCAGTCGTTATTTTTGCCTTCGTATGCGAAGGATT
TTGAAATATATCCGGACAGGCTGCAACCGATCTTCATAAAACTCTTTCTTAAACTGA
GCAAACTGAACAGCATTAGCATTTTGACCCGACCTTCATCGGCACCTGCTGCACA
CCCGCATACGTATTAAAGCTATGTTCGTCTGGCCAGGTTTGCCTTTTTTGGTTGTAA
TCAGGACAACGCCGTTAGCCGCCCGCGATCCGTAGAGCGACGTAGAAAGCCGCAT
CTTTCAGCACGGTCAC SEQ ID NO:182 Late LPZ-122
AGGTGACCGTGAAATATGTGGGAGATGATATGTGGTTTCCTGAATATTCACCTCTT
GTGTAGAAAAGTGAGATCCTTAAGATGTTTTTGCTAATAAGACTCTTAGGAATGTTGG
ACCCCTTTCAGAATGCCATTTGAATAGATTCAAGGTGGTAGCTGTTGCCTGGGGCT
GTTTTAGGGTTTTAGGCCATGCTCTGTAATTCATTGAGTCAAAATTGGATTAACTG
GTGTCTTTTACCTCATAATAGCTACTGCAGTATTTGTCGATATAGCTTCCCTAT- TTAT
TGACTCTCCTTAGGTACGGTCAC SEQ ID NO:183 Late LPZ-124
AGGTGACCGTCCGTCGGGGTGTATTGTCGAACACGTAGGATGGTGCTACGTTG- A
AACCACCGTTACCTTCTTCGATATGTTATAGTTCGAGTTCATACGGAGGGAAT- ACC
GTTTGTAGTGTTATTCAGCACAACCCCGTCCTGATTAAACACCCCCGCAAC- CAAGG
ACGTATTCGACGTTCGGTATTGTTTGACACACTCAAGTATAACTCTGAA- TAGGCGC
TACCCGAAGTAAGCATTGTACCAAGTCGTTATTTTTGCCTTCGTACT- GCGAAGGATT
TTGAAATATATCCGCACAGGCTGCAACTGATCTTCGTAAAACTC- TTTCTTAAACTGA
GCAAACTGAACAGCATCAGCATTTTGACCCGACCTTTCATC- GGCACCTGCTGCACA
CCCGCATACGTATTAAAGCAATGTTCGTCTGGCCAGGTT- TGCCTTTTTTGGTTGTAA
CAGGACAACGCCGTTAGCCGCCGCGATCCGTAGAGC- GACGTAGAAGCCGCATCTT
TCAGCACGGTCAC SEQ ID NO:184 Middle LPZ-126
AGGTGACCGTCGTCAGAAAAAACGTGATTTCCGCAAACTTTGGATCAC- TCGTATCA
ATGGGCAGCTCGTTTGAACGGACTTTCATACTCACAATTGATGCAT- GGTTTGAAG
GGCTGAATCGAAGTGAACCGTAAAATGTTGGCTGACTTGGCTGTT- AACGATGCAGC
AGCTTTCAAACTCTTGCAGACGCAGCTAAAGCTAAGCTTGGGT- AAATAATTAAAAAA
AGAACCGAGGTTTCCTTGGTTCTTTTTTATAACTTTTAAT- GAAAAGTATGAAGAGAG
AAACAGCCTGTCTTCTACTTATAGTATAAGATAAAAG- CTTGTTACTGATAAGACAGC
TTTCATGGTAAAGCAGTTAAAAATAGGGATTTGC- GATATAATAGAAAAAACAGACGT
TTATGTAAATAAAAAACAGTAGAATGGAGAA- ATATGTCAGAGAATCGTTTGGCTTG
GGATCAGTATTTTGCGGCCAGGCTCTCTT- AATCGCTAATCGCTCAACCTGTAAGCG
AGCCAAAGGTGGCTCCGTATTGTCAAG- GATAATAAGGGTTATTTCAACTGGGTACA
ATGGCTCAGTTTCAGGGACTGGAGA- CTGTATTGACCAAGGAGTGCCTGGTCATTGA CGGTCAC
SEQ ID NO:185 Late LPZ-127
AGGTGACCGTGGCGGAGGTTAGGGAAGTTTGACTTCTCATTTTCTC- ACGCACTCCT
CTCCTCGTAACCTCGGTCGAGTCGATGGCGGCTTTTTAGTCGAG- TGTGCTAACGC
CCCTCCGGCCTCAAAATTTCCAGCTACTCGTATTTGATCAATG- CTGAAATCGCGTAA
TTACGTAGTAATAAAGCGTAATGAATTCTATAATGAAGCA- TGTTTCTCTATAGTTCAT
GTGCCGAGAGGAATAATGAAAATGAGGCCTTATATA- TTATCTGGGGCTCAAGGAGA
TGTTATCTTTTCCTCCTTGGTTAGAGACCGTCAA- CCTTCACTTGATTGGATAAAGC
TTCATTTTGTTAAAACCTCCAAGCCAGTAGAT- ACATACGGTAGGCACGTATTATGGT
AGAGACATACGGTCAC SEQ ID NO:186 Late LPZ-128
AGGTGACCGTCCTGTTGCCTTAACCGCGAATCCAAATCGACTT- GGGCTGCTTCCTTT
CGTGCAGATATTTCTGGTTTGGACTCTAGTTCTTGCTCCT- GGAAATCATGCTTGAGT
GCTGGGTAGCTGCCTCCAAGTTTGGTTGACAGGCCCA- TTCCTTACAGCTTCTCTCT
TCCGCTTATGACAGAGTAATGACAGGAATTCAACC- TGACGGATCCGTCTAGCTCTC
ACAAGGTTGGGACCCTGTCTTCGAGAGGGTTAT- TTCTTGAGACTGTTGACTATATT
TGGATGAGCCCTCAGCTCTGTGTACTATTGT- TCATGTACTGGATACTTTGTAAATGA
TTTTATTCTGGTTTTACCCCGGGGGGGG- CATTTTGACTCCTGGGTTTAATACGGTC AC SEQ
ID NO:187 Late LPZ-131
AGGTGACCGTGGAACATGATGATTAGTTCTTCTGTGGGCCAGGATGATTAGTC- TC
TGTGTGACTGTGGGCCAGGATGATTAGTTCTCCTGTGACGACTGTTGGATAG- GATG
ATTCGTCTCCTGTGGACAGGATGATTAGTTCTCCTGTCGAGGCACCCTAC- CCATGC
AATTTGGGATCATGGGAAGTACCTCTCATCTGATCAATGAGTAGGGAA- ATGGGGT
AGGGACCATTAGAGTACTATCGATGGACACATCGTTGTATCTACCGT- CCTATGCTA
GGACGACCTCCATTGTTTGGGATTAGTGAGAGTGGTATGACACTC- TGAGACTGACT
TTGGGTCAGTGGAGGATGTATGATACATCCTCGATCATTTCTT- CTTCTTCATAGTTC
GAGCAGAGCAGAGCACAACAGGCCAAGTAGTGCAGGGTAG- TGCATTTGATGGCTG
GGATAGTAGCGACGGTCAC SEQ ID NO:188 Middle LPZ-133
AGGTGACCGTAAATAAGATGACCCACATGGAGTTTGGCCCTAGTTTCC- AATTTTTAA
CAC1CGCTCTCAACTAGGGAGAACTCCATTCGCTGATCCATTTGT- CCGACTATACTA
TCTCTGCATCAGTGCCCTACACTACTCTGCACTGCTCTGCTC- TACTAAACCATGAA
GAAGAAGAATGACCGAGAATGTCTCATGCCATTCTCTATT- GACCTGAAGTTAGTCC
TATATGAAGAGA1TGTGTCATATCACTCTTATTGACCC- AAAGTCAGTTTTATTGATCC
CAGATCAATATCACAGAGAGTGTCTCAAACCACT- CATACTGATCCCAGATCAGTTTC
ATTGATCCCATATCAAGGAGATCATCCTAGA- ATAGGGAGTACAGTAGATACAATGAT
GCATCCATCAATAGTACTTCTATGGTCC- CTAACCCCATTTCCCTGCTCATTGATCAG
ATGAGAGGTACTTCCGATGAGCCCA- CACTGCATGGGTAGGATGCCTCGACATGAG
AAATAATCATCCTATCCACAGGAG- ACGAATCCTCCTGTCCCACGGTCAC SEQ ID NO:189
Middle LPZ-136
CTAGGGAAGACTTTAAAAGTTTGTAAAACTAAGCATAGCTCTAAACACTGAAGTTA
AAGACATGATTGGAATGTGCAAGTGGTTCAGTATCCAAATATTGAAGGTTGCAGAA
TATGGGCTACTGTGCAAACGAGTAACTTTATCTATATTTTCACAAGATCATACAATG
GGAAACGTGAGATAACAACTGCATCGGTGAACCAGAATAGTTATAAAAGTTCTTGC
AAGTAAAGGGTGAATAATTGCATGGTGTGAATTAAGAATGACCATGTAGAGCTGCT
ATACAGACTTCTCAAGGTTTTATATTTGAGGAGTGCGCGCTATTGATGTTGTGCAAA
AATTTCAGAAATTAATTCTGCGGCATTTATCAAGGTTGTTTGAGCCATTTAAATAGC
AGTTTTTGTTTCTCCAGTACTTTCAGGAAAGCAGGTTAGACGATAAAATGCATC- TTC
CCAATTTACTATATTTCTGTTTTAAAAGATTCTCTCAATGTCCTTAGCACG- TGGCTTT
CATTATTGGGACCAATGAAGATGTGTAGCAGAGGCATTACGTTATGG- AATCTCTCA
CCAAGAACACTGTTTTGGGCTTTAGATAGCTCCTAGTTATAAATG- CTCCAGTGACAA
ACACATCCTAAGTTTGGGGCAATTAATGACGCCTTTTGGTCA- TTCTCCTTTGGGTTT
CAGGCACGGTCAC SEQ ID NO:190 Late LPZ-137
TCCCTTTAGTGAGGGTTAATAGATCTATAGTGTCACCTAAATCGCGGCCGCTC- TAG
AACAGTGGATCCGCAAGCAGGATAGACGGCATATGCATTGGATGCTGAGAA- TTCG
ATATCAACTTATCGATACCGTCGACCTCGAGGG SEQ ID NO:191 Middle LPZ-138
GGTGCGATCCTAAACATGCAAGCTTTGAGTTTGTAACTTTGTAG- AAGTGGACATTTC
TAAGTTGGATGTACAAATCTACTGTTGGTTGTATTGTCATC- CCATAAACAACTGTTT
GATGAGATGTTTTTTTAAAAACCACATCATAATATTTT- TAGGCCTTGTAAAAAAAAAA
AAAAAAAAAAAAA SEQ ID NO:192 Late LPZ-140
ATTCCAAACTTTTCTTCAAGATGTACACCAACATCATTGTCCCCAACTAG- TAGAC
TGACTTTTCACCAGGTCCAAAGAGAGGGGTGGTGGAAGCAGATTTCAGG- CTTTCG
AATAAGTATCAATGATATAAGCATCATCCCCTTGCCAATTGTTCTGGA- TCGCAC SEQ ID
NO:193 All LPZ-141 GGTGCGATCCCATCAGGGGTTGTGT-
TTCTAAGAATCACTTCCATGTTTCAAATTCAG
CACTTGATCTTGTACATACCCAATTTGTTGCCTGCTACTAGCTAGTATTGTCTTTCA
GTTTGAACCATTTTTTTGAGTAAATCGTGTTTAGTCTTTGGCAAAAAAAAAAA SEQ ID NO:194
Middle LPZ-143 GGTGCGATCCGCATTAGAGAAGCATACAGGAAAAAGAAGTACCT-
GCCTCTTGATTT GCGCCCAAGAAGACTCGTGCTATCAGGCGACGCCTTACCAAG-
CATCAGGCATCAT TGAAGACGAGAGACAGTTAAAAGAAAGAGATGTATTTTCCA-
ATGAGAAAGTATGCAG TCAAGGTGTAAGCCACAGGATTTGAGCTTTCATGCAAT-
TTTTTTGTTACTTGCGGGA TGATATTGCCTATATATTTCCGTCCACGTTTTTGG-
CAAATTCCGATTTGCATCAGAA TTCAAGTTATGATAGTGTTCTTCGCTTTTGAG-
CAGTTGATATTGTTTATCTTTTAT TCTCTGAATTGCAACATATTCTAATGCAAT-
GAGTGGATTATATATTGTGGTATTTC CATGTTGAACTCATATAAATGAGCGTAA-
TTGAGTGGTAGCGCTAGGATATTTACAC TTGGCAAAAAAAAAAA SEQ ID NO:195 Middle
LPZ-144 GGTGCGATCCGTATAGGTAGTTTGGATGATGAACGGG- CAAAGAAGGCAAAGGAGT
ACAGGATGGATCCTGTAATTCCTGTTTCAGAAAACA- GAAAATCTGCAATATAAGGAT
GGCTAACTTTTCAGCTATGAAAATATATGGTGC- AGTGGCACTCATATCAGTTGCAG
GTTGTCAAATAACTTTTGTGAATAGGAAAGT- TGTCCTCTTTTAGAGTGCAGAAATCC
TGCAATATAAGATGGCTAAGTTTTTCAG- CTATATGAAAATATATGGTGCAGCAAAAA AAAAATA
SEQ ID NO:196 Late LPZ-145
GGTGCGATCCCATATACAATTACATATATTTTCAACAATTCTTTTG- TTGTTATGAAAA
TCTATTGAAATAAATTGAAATAGTTTGCATCATTTATTTATC- GGAATTCGTATTTATAT
ATTAAATTTCTGATGTCTCAAATCCTTCGTTACTGTA- ACGATATCATTAATATAATGT
GTCTGCAAGTTTATTGGGCAAAACAAAATTTAT- TTTTCGGTCACATCATAAGTTTATT
TTTGGTCACATCATATGCACCATCACATT- AAGCATAAGCATATACAGTAGCGTAAAA
ATACAATTATTGTTGTTGACTAGGAT- CGCAC SEQ ID NO:197 Late LPZ-146
GGTGCGATCCTAGTCAACAACAATA- ATATGTATTTTTACGCTACTGTATATGCTTAT
GCTAATGTGATGGTGCATATGATGTGACCAAAAAATAAACTTATGATGTGACCGAAA
AATAATTTTGTTTTGTCCAATTAGACTTGCTGTATATGTCTGGAGTCCTACCCTTG
AATTGACTTGTTTCCC SEQ ID NO:198 Late LPZ-147
GGTGCGATCCCATATACAATTACTTATATTTTCAACAATTCTTTTGTTGTTATGAAAA
TCTATTGAAATAAATTGAAATAGTTTGCATCATTTATTTATCGGAATTCGTATTTATAT
ATTAAATTTCTGATGTCTCAAATCCTTC SEQ ID NO:199 Late LPZ-148
CCACTGCACCATATATTTTCATATAGCTGAAAAACTTAGCCATCCTTATATTGCAGAT
TTCTGTTTTCTGAAACAGGAATTACAGGATCCATCACTGTACTCCTTTGCCTTCTTT
GCCGTTCATCATCCAAACTACCTATACGGATCGCAC SEQ ID NO:200 All LPZ-149
AGAGCCTTCTTGCAGACAATCCGTGAAAACATGGCTATACAATAAAAATTC- CCAGTT
TGAATTCTAAAGAAAACTGTTCAATATTTGAAGGCCTCTGATATCACA- GAGACTGAT
ATTAAATGGAAATTCATACAAATGAGGAGAGCATGTAGCAACACT- AGAAGCTTTGG
CATAAAGCACCAGATAAATTCATAAGAACTAAATCCATAAGAA- GGATCTCTCGTTCA
CCAGTCACAATCACACTCGGATCGCAC SEQ ID NO:201 Middle LPZ-150
GGTGCGATCCCTGGCCCTGATAACTTTGGTTGCAATGGAAA- ATGCAGTACTAGGTG
CGAAATGCTAAAGCCCGCCCGGAGCGGTGCATGAAGTAC- TGCAATATTTGTTGTAG
TAAATGGCTGGTTGTGTTCCCAGTGGTCACTATGGCA- ACAAGGACGAGTGCCCCT
GCTACAGAGAATGAAGTCCGCAGCCGGCAAGCCCAA- GTGTCCCTGATCTTAGCAC
TTCAGTCCAGTCGCCACTTCTTTTATTCTCTTTTT- TTATAAAAGTGACGAGGCCG
TTCTTGTGCTTGGTGCCATATGTAGAGCGGTGGC- TACTTCTCCTGTGTTAGGAAAT
GTTGCAGTACTAATAATAGAACTTCTT SEQ ID NO:202 Middle LPZ-151
GGTGCGATCCAATAAAGATATACTTTGCAACAA- TAATCAAAATATCATATGCAAAG
TTTAAGATCAAAATAGAATGCAACAAAAAAA- TGGTTGTAACATAGGAACCAACAATG
TTGCATTCAAGTAAGACTCTTTGCAAAA- AAAAAAAATAAAAAAAAAAA SEQ ID NO:203
Middle LPZ-152
GGTGCGATCCACAAGTAAGATAATTGAGTATATATTCAAGATGCAAATATTTCATTA
GGACCACTCATAAAGTTATCAATGATTCACAAAGAGACCTCCTGACCTCTCTCAAAA
GTGGTGGCAACACAAGACTAGTGTAGTTTTTACTATACCTCAATGAAACTACCATCC
TAACTGATGCCATAATCTTCTGTTATATATTACCAAAATTTATGAGATGATTGATCCA
TAAACACTCCAGAACACATAGTCATCCAAAGGAACCTTTGCTTGAATATGGACCCC
CTTAATTCAGGTACTTGCTACTCCAATAAATTGCTTAATCTCTCCACCGATAACCAC
AGTTTGGATCGCC SEQ ID NO:204 Early LPZ-153
GGTGCGATCCAGGACATGAGGCCGAGTTTGCCATTGTGATATGATTGAGGAAGTC
CAGTCTCAAAATTAGGTTTATCTTGATGTTTGACAAGAAATATAGAAGGGCATGATG
AATCAAGAACCTTTTCCAAATCTGTTACTGCAACCAATCCAATGACATAATAACGCC
AATGGTGGTTCCTGTGATGACATAATAAATTGGATTAAATTAATAACATCCCTAATG
CCATGTGGTTAGCTGCATCATCACCGTATCCATCGAGTGTTCAATTTTTGGGAT- GT
TGTATCAAAAAAA SEQ ID NO:205 Early LPZ-154
AAATATTTTTCAATACAACGCCATGTGACATTTTTGTGCTTCTTGTTTTTGATACATA
CTTCCAAAAACTGAACACTCGATGGATACGGTGATGATGCAGCTACAGCCATTGCA
TTACGATGTTACTAAATTAAATCAATTTATTATGTCATCACACGAACCCAAACAATAG
CGCTATATGTCATTAGAATGGTTGCAGTTACAGATCTGGAAACAGATCAATGAA- TCA
TCATGCCCTCTATATCTCTTGTCAAACATCAAGATAAACCTAATTTTGAGG- ACTGGA
CTTCCTCAACATATCACAATGGCAAACTCGGCCTCATGTCCTGGATCG- CAC SEQ ID NO:206
Middle LPZ-155 GGTGCGATCCGTATAGGTAGTTTGG-
ATGATGAACGGGCAAAGAAGGCAAAGGAGT ACAGGATGGATCCTGTAATTCCTG-
TTTCAGAAAACAGAAAATCTGCAATATAAGGAT
GGCTAACTTTTCAGCTATGAAAATATATGGTGCAGTGGCACTCATATCAGTTGCAG
GTTGTGAAATAACTTTTGTGAATAGGAAAGTTTTCCTGTTTTAGAATGCAGAAATCC
TGCAATATAAGATGGCTAAGTTTTCAGCTATATGAAAATATATGGTGCAGCAGAGT
TGTCAATATAAACTTGTGAATAGGGAAGTTTTGGCAAAAAAAAAAAAAAGAAAAAAA AAAA SEQ
ID NO:207 Late LPZ-157
GGTGCGATCCTCGTTGTGAAGACGTAGTGATGGAAAGGTCATGTTTGTAGGAGAC
ATAATTATAGGAGTTTCTTTATTATAATAACCAAGAAGTCCGATCCTGGGGGCGTTG
AGTATATAGTCAGTCTTTGGTAATTTGGTGTGGTGCTGTTTGACCTGCCTTTCCTTT
GGAGCAATGATCCTTGAGGATGGAAGAGGTTATGTTGAGGCTCAAGAGATGATTGT
TTGAGTTGTGGAAAGCAAAAGGTTTCCAGATGTAGTCAGATAGTAACTTCTATGCTT
TTAATAAAATTTAGTCTGTGGGGCATGCCCCTTTTTGCTGGCAAAAAAAAAAA&GAA
AAAAAAAAAA SEQ ID NO:208 Late LPZ-158
GGTGCGATCCGTATAGGTAGTTTGGATGATGAACGGGCAAAGAAGGCAAAGGAGT
ACAGTGATGGATCCTGTAATTCCTGTTTCAGAAAACAGAAAATCTGCAATATAAGGA
TGGCTAAGCTTTTCAGCTATGAAATATATGGTGCAGTGGCACTCATATCAGTTGCA
GAGTTGTGAATATAACTTTTGTGAATAGGAAAGTTTTCCTGTTTTAGAATGCAGAAA
TCCTGCAATATAAGGATGGCTAAGTTTTTCAGCTATATGAAAATATATGGTGCAGCA
GAGTTGGAAAAAAAAAAAAAAAAAAAAAAAAAAA SEQ ID NO:209 Middle LPZ-162
GGTGCGATCCCAGGAGAATATTAGTTTCATGTGTTGCTATCATTTTCTTCA- ATATGC
AGGGCAACCATTTGAATGAAACTATTCCTTTCGAATTTCAAAAACTTA- ATAGGCTAA
CTTATCTATCTGGAGCCGATTTTCATTGACGAGTAACCTGTAAGC- TGGCCAGCAAA
AGCCAACAGATGTTCAGCTTGTTGGAACCAGTTGAAGATTGTA- ATAGAGATGGTGA
ATAATCGCGGACGGCTCGGCCAATGGAATATTTGTTGCATC- ATCATCAAGGGGGTA
TGAATTCCAAAGAACTTGTTGATTGAAATTCCCAAGCAA- AATTCTGTGAAATGAAAA
ATTTATTGAGACCATTGGGCAAAAAAAAAAAAAAAT- AAAAAAAAAAAAAA SEQ ID NO:210
All LPZ-165
GGTGCGATCCGACTGTGATATGTGACTGGTGAACGAGAGATCCTTCTTATGAATTA
ATCTGGTATCTTTATGCGAAAGCTTCTAGGGTTGCTACATGCTTCCATTCTAATATC
AGTCTCTGTGATATCAGAGGCCTTCAAATATTGAACAGTTTTCTTTAGAATTCCAAA
CTGGGAATTTTTATTGTATAGCCATGTTTTCACGGATTGTCTGCAAGAAGGCTCTTT
GGCAAAAAAAAAAAA SEQ ID NO:211 M,L LPZ-166
TTTTTTTATTTTTTTTTTTTCCAACGAGATCACTGTCATTGTTCAATAACTATATGCCA
AAGAGCCTTCTTGCAGACAATCCGTGAAAACATGGCTATACAATAAAAATTCCCAGT
TTGGAATTCTAAAGAAAACTGTTCAATATTTGAAGGCCTCTGATATCCCAGAGACTG
ATATTAGAATGGAAATTCATACAAATGAGGAGAGCATGTAGCAACACTAGAAGCTTT
GGCATAAAGACACCAGATAAATTCATAAGAACTAAATCCATAAGAAGGATCTCTCGT
TCACCAGTCACATATCATACTCGGATCGCACC SEQ ID NO:212 Middle LPZ-167
GGTGCGATCCGACTGTGATATGTGGCTGGTGAACGAGAGATCCTTCTTATG- AATA
ATCTGGTATCTTTATGCGAAAGCTTTTAGGGTTGCTACATGCTCTCCTCT- TTTGTAT
GAATTTCCATTCTAATATCAGTCTCTGTGATATCAGAGGCCTTCAAA- TATTGAACAG
TTTTATTTAGAATCCAAACTGGGAATTTTATTGTATAGCAATGT- TTTCACGGATTGTC
TGCAAGAAGGCTCTTTGGAAAAAAAAAAAATAAAAAAAAA- AAA SEQ ID NO:213 Middle
LPZ-169 TCCCAAAGGCAATTATACATGGATC- GCACC SEQ ID NO:214 All LPZ-170
GGTGCGATCCCCACTGCAGAAAGATG- AGCCAGTACCCTGAAATTTTGCTGTTGTCC
ATGCCTGGGTCACGGAGGAAAGAA- CGGCACGGTGCAATATGATTTTGCTACATACA
AGTTCCAAGAGTGGATGCAGACAGTGCTGGCCATGGCTGATTATTTGCAGGTGACT
AATGCTCTTTTGGTTATCCTTACCATCATCATCTTCCTGCCATTCTTTTGTACCTCGG
TATGGAGACGAACACCCACTTTTCAAAGTTTGCAGAGGAAGCATGTATTCATAACA
GGAGGATCAAGCGGCATTGGCCTTGAGATTGCCAAAGAGGCTCTTCACAGGGTT
CTTACGTGACACTGGCGTCAAGAAATCTTTCTAAACTTCGTAGGGCTGTTTGAAGAA
ATCATCCAAGAAGTGGAGTGCGACGGAGACAAGATTAATATCAAGGTAATATACCC
TGCAAAATGTTGTCTGGAATACAATCCAAAACCAATTTAGCAATTAACCCATTGGCA
AAAAAAAAAAA SEQ ID NO:215 All LPZ-171
GGTGCGATCCAAGTGCGGTATTCTTCCTTTGGCAGTTCTCTGAACTGTTGAGAGAA
TTTGAGTAGGATAACGACATAATTACTATGCTCACAAGCCCAGACAACACGAATAG
ACTCCCTTCCGTGCGTCGCCTTCCAGAGGACGCAGCAGCTAAAATCTCGGCCTGA
CTCACCACATATATATTTAATAGCTTGTATATGCCATATGAACTGTTAGCATGATCTC
CCTCTAACTGCGAATTGTGTTGCTGTAAACTAATCCCAAAGGATGTTTACTCTGTTG
CTTTTCCAACTGCTGATGGATTTCGCTCATACAATGACCCGAGAGCACCATAAACC
ACCCAGCGTTGTGGCCTATGACCCATAGCTTTTTGTTCGCACAGCAATTGAAGACC
GGCTACAGGAGATGACTAATGCACTTCCGAGAAGGTTTCACCGCGAATGACAGGG
AAGGACAAGGCAGAGCAGCAGGCCAAGACAGCTTTAGTCGCAGAAGTTCAAGCA- G
ATCTAGATTCATAGTAAATGGAAGTTCTACACTAGTTACAAATTTAAAAACGT- ACCTG
CATGGACTACACGGTTTATTTACGAGTGCCACTTGTCTCATTGTTTTCC- ATCAGATG
TCTGCTGGATTGTGGTAGTGTGTTCTACCGTATCGGTGCGGGTTTT- GTATATTGTG
CGTCGACAGAGTGACAGGTGGTGATTTTACTGGCAATTAAAAAA- AAAAAACAAAAAAAAA A
SEQ ID NO:216 Late LPZ-172
GGTGCGATCCTAGTACAGGCGTTTGGAACAGAGTGGAGAATATGTGGAGTATTGG
GGGATGCCCCCGGTCGTGTGTTGCTGCGTTTGGGAATTTGTATTTCTTCCATAGGC
AACAAGTGATGTGTTATAATAGTAAAGAGAATGTTTGGGAAGTGGTGGCATCTCTTC
CTGGAGACATGAATATTGTTACTTTGCGCAACAGTGTGGTGTGACAAGATATTTGT
GAGCGGTTGTGCTTGCAGTGGCGGCGATCAGGTGTGTTACATGCTGGACAAATCT
TGGGCGTGGGCTCCTATTGAGAGGTCACATGAGTTTGAGGGTTTTGCTCAGTCT- G
CAATAACTGTAGAGATATGAGCAAATTCTGTTGGGTTCACTTAATTTTGGGAT- TATT
ATAGTGCAGAGGGGAGCCGGGAAGTTTCAGTGTACAGTGATGGGCACCAC- ATGTT
GCCAGCATTGGGGGTGCCCTGTGAATATGATTTCTATAAGTCCGGATTT- TAAATATC
TAGGCCATCTATCTCATCCAGCCTCTGATTGTGTCTGTACTAAATA- TATCCTGTATA
TTCGTGATCCCTGGTTTTGAAGTGAGCAAGTTTTAGTGGAAGA- GGATTTTTATTAAA
TATATATAAAGTTTCTGTATTCAGGGTTTTGGCAAAAAAA- AAAAAAA SEQ ID NO:217
Middle LPZ-173
GGTGCAATCCGCCATAAGAGAGGCATACAGGAAAAAGAAGTACCTGCCTCTTGATT
TGCGTCCCAAGAAGACTCGTGCTATCAGGTGACGCCTTACCAAGCATCAGGCATC
TTGAAGACTGAGAGACAGAAAAAGAAAGAGATGTATTTCCAATGAGAAAGTATGC
AGTCAAGGTGTAAAGCCATAGGATTGAGCTTTCATGCAATTTTTTTGTTACTTGCG
GGATGATATTGCCTATTATATTTCCGTCCACGTTTTTGGCAAATTCCGATTTGCATC
AGAATTCAAGTTATGATAGGTGTTCTTCGCTTTGAGCAGTTGATATTGTTTATCTT
TATTTCTCTTGAATTGCGAACATATTCTAATGCAATGAGTGGATTATTATATTGTGGC
AAAAAAAAAAAAAAAAAAAAAAAA SEQ ID NO:218 Middle LPZ-174
GCGGACGCCTCAGGATAGCGTTAGGGTTGCCTTAGGATAGCGTTAGCTCTGCCTT
CTAAGGTTGCCGTCTTATCCTCCAGCGTCTAGGGCTTCCACTCCTAGGATTTCTCT
TCCACTAAAACCCAAGACAAGTGGAGAGAAATCAAGATAGAAGTGTGTGTGAAA- TG
ACTCTTAAGTCATCTCCTTTTAGACTAAAACATTGAGCACATGTGGGGTTTA- TTTGG
TTGCTGGCCGTCGTT SEQ ID NO:219 Middle LPZ-175
GGTGCGATCCTGAAACAACATATTCCCGATGGCTCTTCCGAAGGAACCATTGCTCT
ACTGTGTGGCCCTCCCCCCATGATCCAAGATGCCTGCCTACCTAACCTGGCCAAAA
TGAATTATGACATTCAGAATTCGTGTTTTCAGTTCTAATTACACCCTTCTGGTT- AATC
AAATTGGGACATCCCCTCCCACATCCTGTTATTAATTAAGCCATAGTCTA- GTGTATA
AAATCTGTTGATGTGTACAGCATCAAGTTAATTTCCTCCTTTTCTGT- CAAAAAAAAAA
AAAAATAAAAAAAAAAAA SEQ ID NO:220 Late LPZ-177
GGTGCGATCCGATCCTAAGCGGGTGCATATATATAATGACAAGCTGTAGTAAC- TAA
CTCTTGTCATGAGGCCATTGCTAACATAGCCTGTCCAATGCACATAGCAGT- CAAAA
AAAGCAAATAGCCGCCATGTTCCCATACACGAAGTAAGTACCCTCCCTA- TTGAGTC
ACCTTACCCGCCGAGAGAGATCCCAATTCCATGTATTCGGTTAAGTA- AGCCCTGCC
AGCTATGTCCCACCCATGAAAGAAAGTACTGATCCGAGTGGATCG- CACC SEQ ID NO:221
Late LPZ-179 GGTGCGATCCAAACTGTGGTATCGGT-
GGAGAGATTAAGCAATTTATTGGAGTAGC AAGTACGCTGAATTAAGGGGGTCCA-
TATTCAAGCAAAGGTTCCTTTGGATGACTAT GTGTTCTGGAAGTGTTTATGGAT-
CAATCATCTCATAAATTTTGGTAATATATAACAGA
AGATTATGGCATCCAGTTAGGATGGTAGTTTCATTGAGGTATAGTAAAAACTACACT
AAGTCTTGTGTTGCCACCCACTTTTGAGAGAGGTCAGGAGGTCTCTTTGTGAATCA
TTGATAACTTTATGAGTGGTACCTAATGAAATATTTGCATCTTGAATATATACTCAAT
TGATCTTACTTGTGGATCGCACC SEQ ID NO:222 Late LPZ-181
CAATCTGTCTGCAATTGATATTATTGCATCCAGTAAACCAGATACACATTCACCACA
ACATTAGAGACTCTAGAAGTTCCTTTGGCGACAGGCAAAACTCATGATTACAGATAA
TTGGAGTTTCCTCTAACCAGAGTCAAACGATCTAAAGGGATTTGTCTAGTCCTCCAT
TCCCTCATTCAATGAGGCGATGGCTTATGCCGTGACAACAGTTTCTATAGTTGCAT
CCGCTCCTGTTGATCCCACAACATTTTTGGTGTTCTCTGCATCTTCTTCCTCCC- ATA
TCTCTGGCAGGGCTTGTCTAATGTTGTGAATACTTGCAAGGGCAAAATCTG- CTCCC
TCTGTTCGGATCGCACC SEQ ID NO:223 Late LPZ-182
GGTGCGATCCTCTCAGTTACGAGCTCAATTTCGACCAGGGGTCTCGGCAAATTGA
GGATCATGAGAAGCAGGGTATGCCCTTGAATGCCCTGAAGCCAGGGGAGTCTCAG
GGCAATCACGAATGAAACCTGACAAACCCTAAGAAAACCCCTAGAGCGTGCCCTGC
AGAAAGGGAATTCTTTTTGAGGCCGGCGGTCTTTCTGTCGTCTTCTCGCAGCCGTA SEQ ID
NO:224 Late LPZ-186 GGTGCGATCCAGCAAGAGAACGAAAAAGGT-
ATGAGAATCTATGAAATATTTGTACA TCACTGTATTCATATGAGGGCCTTTTTT-
TACAATGCGGTAGGGTTGTTTGGAGAAT AGAACCTGATTAAAATGTAGATGGAT-
TCAAGCTTTTAGTGAAATGAGGCTCGGAAC GCAAGTATGCTGTCCACTTTGAGA-
CTCATTCTTCTATAGTATCTGAAGCCAAAGCC SEQ ID NO:225 Middle LPZ-189
GGTGCGATCCCATGGGATAGTTGCAAAACACACAAATTTGTTGTGAAAGAAGAGAG
ACACGCACAGACAACCATATGATCTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTT
TTTTTTTTCACAACTCTGCTGCACCATATATTTTCATATAGCTGAAAAACTTAGCCA- T
CCTTATATTGCAGGATTTCCGCATTCTAAAACAGGAAAACTTTCCTATTCACA- AAAG
TTATATTCACAACTCTGCAACTGATATGAGTGCCACTGCACCATATATTT- TCATAGC
TGAAAAGCTTAGCCAGCCTTATATGCAGATTTTCTGTTTTCTGAAAC- AGGAATTAC
AGGATCCATCACTGTACTCCTTTGCCTTCCTTGCCCGTTCATCAT- CCAAACTACTAT
ACGGATCGCACCA SEQ ID NO:226 Late LPZ-194
GGTGCGATCCTGCGAGAGCCGAGGGTTCATTTTCCTTTCGACAACGACGTTCA- GT
GGCGACCAGAGTTTCCCAATCACTTCAGCGATTCTATTCCTTCGTTGTAATA- AAGCT
TAAGGAATCCATGCTTTATTCCTTGGAAGGTTTGAATATTTATATTTGT- TGGCATTAA
TGCTATATACATCTATACTAATTTTGGGTTGTTCTAAACTTGTTT- TGAATAACTTAAA SEQ ID
NO:227 All LPZ-195
GGTGCGATCCATGGCAAAGAGCTCGTTCAAGCACGATCATCCTCCAGAGAGAAGA
CAAGCTGAAGCTTCTCGGATTCGAGAAAAGTATCCGGACAGGATTCCGGTTATTGT
GGAGAAGGCTGAGAGAAGTGAGATACCTGATATTGATAAAAAGAAATATTTAGTCC
CAGCAGATTTGACTGTTGGGCAATTTGTTTATGTTGTCCGAAAAAAAAAAAA SEQ ID NO:228
Middle LPZ-196 GGTGCGATCCCCTGTATTCTTGAAAGGGTTATAACGGAAGATAG-
CATTTTGCTCAG ATTGTAGACAGTCTGCATGATTTGTCAATACTACTATTTCGC-
ATTATTTGTTAATACT ACTAATCCTTGTACTCATCTAGACTATTTAATTATTAA-
ATTCTACAGTTTCTTTCTCCT AGATGGCAAACAATATGAATAAAATGCCAATAG-
TTTTGGAACTACTCCATTAAGAGC TTTAGATGATTATCATTCATCATTTGCCTG-
TTTTGAATCGTAAATGAATGTGTCACGG TCTTCTTTTCTGTTAGTCTCTATGCT-
TTCATCAGAAGAGTCTAAGCCAGTTACTGGA AGCTATTTGTCATCTCTTTAAAC-
ATTGTTTCCGTGCCAAAAAAAAAAAAAAAAAAAAA AA SEQ ID NO:229 Late LPZ-197
GGCAGAACTTCCAAAGTCTAGTATTTGATTAACTAATATGATGAAG- ACACTCAGTCT
ATAACATGACGCCAGAAATCAGACCATATGCATGATAACTAGC- ACGATTAAAATACA
ATTCGCAACCTTTAATACACTAAAAACGTTTACTGTATAG- TCCACTCAGAACATTTC
GATAGTATTGTCAGATCGACTTATTTAGCTCATATTC- AGCAATCTGAACTGTACGAT
GCGGCTCATTCAAGGGCATTTGGGTTTGCCCTTG- GCATTCTTCATATCCCGATAGC
AAGGACACGCGTTCTTGTTGCCATATGTCCCT- GGGGGATCGCACC SEQ ID NO:230 Early
LPZ-198 GGTGCGATCCACATTGGCCAGGCCGGTATTCAGGTCGGCAATGCCTGTTGGGAGC
TTTACTGTCTCGAGCACGACATTCAGCCTGATGGACAAATGCCAAGTGACAAGACC
GTTGGCGGTGGAGATGATGCATTCAACACATTTTTCAGTGAGACAGGTGCCGGTAA
GCATGTTCCTCGT181GCCGTGTTTCTGGATCTGGAGCCAACTGTCATTGATGAAGT
TCGAACCGGCACATATCGGCAGCTTTTTCACCCAGAGCAGCTGATCAGTGGCAAA
GAAGATGCCGCGAACAACTTTGCTCGTGGCCATTATACCATTGGTAAGGAAATTGT
GGATCTGTGCTTGGATCGCAGC SEQ ID NO:231 Late LPZ-199
GGTGCGATCCCAGCATTGGATGCATTTCTAGCACAAAGCCATCTTGACTAAAATAG
CACTGCGGGCAACTGCAGTCCATAACTTTCAGAGCATTGTTGCTGCCTCAATTGTA
TACCAATCCATATTCTAAAAATTAGACCTGGAAACCAGTCAGAAATTTAATGTTTTCT
TGCAGAAAATGCCCTTTTAGAAAAAGGAGAGAATAACTGCATTCAAGTTCTAACTCC
CAGACATAGCCTGGCAACGTCATTCATCAGTTCGGATCGCACC SEQ ID NO:232 E,L
LPZ-201 GGTGCGATCCAGAAAACAGCACAAGCAATCTGTAAGACCAAT- ATTATTATCATCTCT
CACTGCTCGTGAACAAAATGCTGGTTCATAGCCATCACG- AAGGCTAAGGCTACTAT
CCAGCCAAACTGATCTOCAACAATAATTTCATAAGCT- TAAATAAATAGTCCATCCAG
TGGATGGAGCCAGAAAGCCATAGAAACTTCAAAT- ACTTGTGGTATCAATCTCTCCTC
TGTTAAGGGAGGTATCAGATCAGAAGCACTA- ATCAAATGCATACATAAATGCAGTA
GACTGCAATAAAACAAAATCTGCAGATAG- CAACTGAGCGCTTAACGAACGGAAAAG
AGTTTAACTTGATCTATCACAGGATCG- CACC SEQ ID NO:233 Late LPZ-202
GAAAATGGGAGCCTCAAATATTCAAA- GCCTCATCTCAAGAGTCTCAGATTCGGATT
CATTTCATTTGGTTCGTAATAAAA- TAATGCATCAAATAGTTATTATCCACAAAAATGG
GAGAATTATTACAATCTGTCTTCTCAACATAAAGTCATAGCATAGCATAGAACCACA
CCACAGTCGTCATCATTTGTTTTGTTCACCACCGAAGGGGCTCTTTACAGCGTCCA
TGAAGCCCTGTGTAGCACCCTTCGCCTTGTCCCCCGCCTGTTGGAAGAAAGAGCC
AGTTTGTTCTTTCCCCTCTTGGGCTTTTCCCGTGATGGATCGCACC SEQ ID NO:234 Late
LPZ-203 GGTGCGATCCTATTATAGAACCATGACTCTTGTCGATGGGGCATAAACTT- CTCATTC
TTAGGCGTGCCTACTGTGACTCTTGCCGATGTGGCATAAACTGCTTA- TTCTTAGTT
GTGCCTTCTGTGCAGAACTTGTTGAGTCGGTGGATTACACTGAC SEQ ID NO:235 Late
LPZ-204 GGTGCGATCCATTAACTAGATTAACGATAA- CATTCCTCTGCATCCAATCCAATGCTC
ATCTAAATCTACTTCTACTTAGATCTC- TGCCTCATCTTTCTCCACCTCCTCATCCATT
CTGAAATATTAATTTCTGCATAG- ATTTTGTTAGGGTCTAGTAATCATTTTCATGAATT
TAAATCTGTTCTAGTCTCTTATTATTATGCTGCTTATGCTAGCATCAGAACCTGTGTA
TAATTCATTCATGTATATATTGGATTACACAAATTATACGGATGCCAGAAAAAAAAAA
AAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAA SEQ ID NO:236 Late
LPZ-205 CTTGAAGCTGATATGTTTGAACCCGAAATTTTGTTACCCAACTCCAGTGT- ACATTGT
GTCACTGTCAAAGAGAACATGAGAGCTGCATGCAAGCTTTTGCATGA- TAGATAGAT
TACTGATCACCGAACATTTCTTACTCTACTTTCCTCTCCTATCCC- CAGTGATTTTTG
GGCATTTTCTATACCCTTCGGATCGCACC SEQ ID NO:237 Late LPZ-206
CTCATGAACAGCAATATGATGCATTCCTCTTATACACATTTCA- TATATGTCACCCTT
GCCGTCATGGCTACTCTAAGAAGAGCAAAACAGACCCATT- GAATCTTTACACGCGC
TTGTTTATATGAATACAAATAATTTAGGCGTTTCTTTA- CACGCCCTTGTTTACATTAA
TACAAGTGATTTAGGCGTTGTTACCAGAATAGTG- CCACGGATCGCACC SEQ ID NO:238
All LPZ-207 GGTGCGATCCCAAGATAGAAAAGGGAACTATGGTCTCGAGGAGTGTCAGGTGCTA
CAGATCACAATATACATAAGGGTCTGATAGTAGTACTCGGCCCAATGTTTGAGGGC
TCTAACTAAGGAGGATCAACCGTACCCTTAGCCGTAAAACCCGACTACCCTATCGT
ACGGGCGAGTAATCTCTCTGAGTGTTGTTCTCGGTGTATCGTAGCAGCAACACGG
CTGACGGTTTATCTATGGTGAGGTTTCAAAGGAGCTAGGGGGCTTCCAATATACCC
AGAGGGTACTTGGAAGACAGTTTATACGCGGTTCTGTCTAATGCGCTACTACTCGA
AGGGGTACCCACAGGGGTACAAGAGAGTGCAACAAGCATGACCACCCCTTGTAT
TTCTTGCATGTATGCCTCCCCAAATCCGCAGGTTTATGCGCTCATTGACAGATTCC
GTGGTTTAAAGATGCCGGAACATGTCTCTAGCCAAAAAAAAAAAAAAAAAAAAAAAA
AAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAA SEQ ID NO:239
E,L LPZ-208 GGTGCGATCCTCCTAACCTGCAATGTCCTTCCTGCA- ATTATCAACAGA
AATTAGGTTTATTTTTCTTTTTGTCTTTTCTTCTTTTTTTTT- TTTTTTTTTTTTTTTTTTT
TTTTTTTTTAAGTAAACGACCATTTCAAACGCCAT- TCAAATGCTATGAATTAATGTT
GAATTAATGTTAGCATTAAGTCTTTTAACATT- TTATGTAAGGCATATATATCGTTCCA
ACTACTCTTACAATACACCTGCGGTGTA- CTCCTGCCACCGCATGTACCACCGTTAC
ATGTACGCCTGCCAGCACATCTAACA- GGTGCCAACTCCTTTGAACTCATCGTCGCC
ATTTTTGTATGCATATTTGAACTC- ATCGTCGCCATTTTGGTATCTTCACATATGGCC
AGTCCAGGATCGCACC SEQ ID NO:240 Late LPZ-210
GGTGCGATCCAAGGAGTGGGCGTGCAATGCGTCGA- AGATAGCCACCACTGCAGG
GGCGTGGCATGCTGCCGTGCTTCCCACAGGGAGAT- CAACACCTGCACCTCCGCCT
CCTTCCGCGGTTACCACGAG SEQ ID NO:241 Middle LPZ-211
GGTGCGATCCAGCCACAGAAAGATTGGTTTACTCGATAATTG- AACGGTAGACTTTG
TGCAGGTTTAGATTGTGTACATGCTGATCAGTATTGTCTA- CACCATTTTCAATCTTG
TTTAGTTCTATGGTAATTTATGTAACAAATTCAGCGA- TGTTGGGGAAATTGGTCACA
TCAGCTTTGTGCCTATATATTCAAGTAAATCAGG- GGATCCATTAATACTGCTTTT
AATAATTGGGGCAAAGTTGTGGGATGACTGCTT- CAGCGGAATACGTGCTTTTCATA
GTGCTGTATGACATTTTGTTGAATATGAATT- TCTTTGTGATACAGTTGCGCGAAAA AAAAAAAA
SEQ ID NO:242 Middle LPZ-212
GGTGCGATCCATGCCAAGAGGGTGACCATCATGCCCAAGGACATCAGC- TCGCTC
GCCGCATCCGTGGAGAGAGGGCATAAACAGTCAGTCAGATCCAATGGT- GTGTTTT
CACACCACCATATGTTTCTTTTACTAAATTTGTTAGGTCCCTTCGGT- GGGTCTTTTC
TTTCCCCCGATTTTAGTATTTTGTTGTTCTTCTGAGTTTCATCA- TTGCAAGTACAAGA
TGCAGAATTGATGGTTATTGGGACTTGGAGACTGGTTATT- GCTATGTAGAGTATTTA
TATTAGACAGGTTTCACTTGAAGATATAAAATTG SEQ ID NO:243 Late LPZ-213
GGTGCGATCCTCATGTGTATAACCGAAGTTTGC- GGGATTCAGATGGTCAGTATCT
TAAATGTCCAACTTTCGGTACGAATGGGGTGC- GTTCTGAAACGTGCCACGAAAGAG
GTGTTCAGGATCTGTCTGAGGCATCTTTCC- GGTATTTTCCACTTCCATGGTATGAG
AAACTTTCGTCTTGTTGCAG SEQ ID NO:244 Late LPZ-214
AGGAGACACAACTTTACGAAAAAGTTCAATCTGGAGTC- TTCTAAGTTTTTCAGACTC
TCTAAATATGAAAAGCGCCGAGTTTCTCCTATACT- GGACTCGTTAAAATTTTACAGT
AAAGGACCTGTTCTATTACAAACAGGAACGGA- CCGCTCCTCCTTAGGGATCGCACC SEQ ID
NO:245 Late LPZ-215
GGTGCGATCCAGCAAGAGAACGAAAAAGATATGAAGAATCTATGAAATATTTGTAC
ATCACTGTATTCATATGAGGGCCTTTTTTTACAATGCGGTAGGGTTGTTTGGAGAAT
TAGAACCTGATTAAAATGTAGATGGATTCAAGCTTTTAGTGAAATGAGGCT SEQ ID NO:246
Late LPZ-216 CTCAACATAAAGTCATAGCATAGCACCACACCACAGTCGTCATCAT-
TTGTTTTGTTC ACCACCGAAGGGGCTCTTTACAGCGTCCTTGAAGCCCTGTATA-
GCACCCTTCGCCT TGTCCCCCGCCTGTTGGAAGAAAGAGCCAGTTTGTTCTTTC-
CCCTCTTGGGCTTTT CCCGTGATGGATCGCACC SEQ ID NO:247 Middle LPZ-217
GGTGCGATCCCATGGGATAGTTGCAAAACACACAAATTTGTTGTGAAA- GAAGAGAG
ACACGCACAGACAACCATATGATCTTTTTTTTTTTTTTTTTTTTTT- TTTTTTTTTTTTTT
TTTTTCGGGACCAAATATTTTTCAATACAACGCCATGTGA- CATTTTTGTGCTTCTTGT
TTTTGATACATACATTCCAAAAACTGAACACTCGAT- GGATACGGTGATGATGCAGCT
ACAGCCATTGCATTACAGATGTTATTAAATTAA- ATCAATTTATTATGTCATCACACCA
ACCCAAACAATAGCGCTATTATGTCATAG- AATGGTTGCAGTTACAAGATCTGCAAA
CAGATCAATGAATCATCATGCCCCTCT- ATATCTCTTGTCAAACATCAAGATAAACCT
AATTTTAGGACTGGACTTCCTCAA- TCATATCACAATGGCAAACTCAGCCTCATGTCC SEQ ID
NO:248 Late LPZ-219
GGTGCGATCCTGGACTGGCCATATGTGAAGATAACAAAAATGGCGACGATGAGTTC
AAATATGCATAGAATAAGCGTTCTGTAATTGGAACGGCCATAGGAGTTGGCACCTG
TTAGATGTGCTGGCAGGCGTACATGTAAACGGTGGTACATGCGGTGGCAGGAGTAC
ACCGCAGGTGTATTGTAAGAGTAGTTGGAACGATATATATGCCTTAACATAAAATGT
TTAAGACTTAATGCTAACATTAATCAACATTAATTCATAG SEQ ID NO:249 E,L LPZ-220
GGTGCGATCCCATGGGATAGTTGCAAAACACACAAATTTGTTGTGAA- AGAAGAGAG
ACACGCACAGACAACCATATGATCTTTTTTTTTTTTTTTTTTTTT- TTTTTTTTTTTTTTT
TTTTTTTTTTTTTTTTTTTTTTTTTTTTTGTTTTTTTTT- TTTTGTGAAGTGACAAAATCTA
AACCAAAGATTAAAAGGCTTTGGCTTCAGATA- CTATAGAAGAATGAGTCTCAAAGTG
GACAGCATACTTGCGTTCCGAGCCTCATT- TCACTAAAAGCTTGAATCCATCTACATT
TTAATCAGGTTCTAATTCTCCAAACA- ACCCTACCGCATTGTAAAAAAAGGCCCTCAT
ATGAATACAGTGATGTACAAATA- TTTCATAGATTCTCATATCTTTTTCGTTCTCTTGC
TGGATCGCACC SEQ ID NO:250 Late LPZ-221
GGTGCGATCCCAACCAGGTGTCCATGCAATATATGGTG- AGCATCAAGTTTGAGGTG
GTTGATTGAAAGTTACAAATTGGTGACATCTGAAGT- CTCATTCAGTTATGTTTTTGT
TATAAAAACCATAACCAATTTTGTATATAAGAT- CCATAATCAATTTTGGCCAA SEQ ID
NO:251 Late LPZ-222
GTTTTCAAGAAGAGCCTGACGGTTTCCTCGGCGGGATGACGGAAACAGGAAGCGG
CCGGCCGGTTCCGGACCCTCCGCAGGCGGAGCATAGCATTTTGCCGGAACCACC
GCATGTCGTGCACCCAACATCCGCGTCTGACCAGCGGAGGCACATGCACCCAACC
CTCCCGGTTCCATGCACCTCGGGCAGCGCGGCCACCCGCCGGCCATCGGCTTAT
CCATCATGGATCGCACC SEQ ID NO:252 Late LPZ-223
TGGGCGAATCATATGGCTTGCATTTTCATTGTAACATGTATACGTTAAGGATTATCA
TAATGCCTCCAAAACCTTGTATCTTCGTCCTTGCCACAATACATCCAGGATAACTAA
TGGAAGCTTGACATGTCTTCACCAGTAATAATATATCAACTATAATACATGCCATTC
TTTATCAGTTTTGAACAAAATAATCGATTTGCATTCTTGACAAAGAACCTCGCGCAT
AAAAACAAATAAATTCTCATAATGCCTCCCAAACCTTGTAGTCTGGGCCCTCAGTCG
CCACAATCCATTTAAGAGGAATTTGGGGGTTGATAGTGCCCAGGTCCAATCTTCAT
GAAAATTCGTTCATCAATCTTTGCTGCATACACATCTCTCTCTGCTTTCACTAT- CTG
GGATCGCACC SEQ ID NO:253 Late LPZ-224
CCACTATAATGAACATTGATATTACAAATATAATATACATAATATTACAATTCAAATC
ATTGACAATGAGCAGGCACTACTTGCAGTGCTTTGGAATTCAGACTTCTGATTTGC
ATTAATTCTTGTAGACGCTTTTCTGGGAGGGCAGGTTTTCCGCTTCAGAGAAAACC
ACGTACAAAACGATATTAAATAAAAATAGACACATACAAAAAATACTTCATTTTTTG- C
TCTTTCCATTTGGTTTCTTCCTCTATCTCCATTTTGGAGGGCTTAAATGACTC- AAAT
TTAAAAGTCAACAACAGAGTGCAGCACATTCTATTAGCTTTGCTGTAAAT- ATCTGAT
TGGATCGCACC SEQ ID NO:254 Middle LPZ-225
GGTGCGATCCGCATTAAGAGAAGCATACAAGAAAAAGAAGTACCTGCCTCTTGATT
TGCGTCCCAAGAAGACTCGTGCTATCAGGCGACGCCTTACCAAGCATCAGGCATC
ATTGAAGACTGAGAGACAGAAAAAGAAAGAGATGTATTTTCCAATGAGAAAGTA- TG
CAGCCAAGGTGTAAAGCACAGGATTTGAGCTTTCATGCAATTTTTTTGTTAC- TCGCG
GGATGATATTGCCTATTATATTTCCGTCCAAGTTTTTGGCTAAATTCCT- ATTTGCATCA
GAATTCAAGTTATGATAGGTGTTCTTTCGTTTTTGAGCAGTTGA-
TATTGTTTATCTTT TATTTCTATTATTAATCTTCTAAGTTGGATCGCAC SEQ ID NO:255
Late LPZ-226 AAACAGACAAATATAGAAATATGCATACATAAGTC-
CCTGCAGAATTGTTTTCCGCAA TGAATTCTGGTTTATGGCAACATTACCTACTT-
AGTACTAACCCTAAGATTATTTTCAG CTCTGATAAGTGGCATACGTGTATCAAT-
CTTGCATGAGTCTATCCCTGTTTTAATCT TTTGTTGGGATCGCACC SEQ ID NO:256 Late
LPZ-227 GTGGAAGCTTCATTGTAAAACACTACTGGTTTTGAGAG- AACAAAATATATACGCTAG
CCGAGTGGATTATAACAAAATATAGGCTTTATTTC- TATTGGATCGCACC SEQ ID NO:257
Late LPZ-228
GGTGCGATCCCATACATTAACATAGCCATCACAGCCCCCAGTGGCAAAAGTACCAT
AGCTGCAAAACATTATAAAACTAACATTCCTACAAGGAAATAAAATACAACTAAAAA
AGCAAGCAATAGGCATTAGGGGAGGGAGAAGCTAAAACTATTAAGCAACTTACATG
GGATGAAAGGCAATTGCGTTTACTGGATAAACAGTATCTCTGCCAGCCTCTGACTT
GCGATGACATTTAAAGGCATATTTTTTAAGCTTGACCAGCTTCAGATACATCATAAT
ACTCCATAGCCATGCGAGCTTCCACAGAACTAAGGGGCAAAACCTGTCCATTGG ATCGCATCA
SEQ ID NO:258 Late LPZ-231
GGTGCGATCCAACTGAGAAGGGTGTTGGTGGAAAGATGACACCAAGTGGGTTCT
CTATTCTCCAGAGGATGCAAGAAAAATTCTGAGAGCAAAGAAGAATGGGGACTCAA
ATATTACGTTGGGTTCTGTTAAATCTGCCAAGTACCCTTCAGGAAAGCTTTATGCCA
TAGACCTGGTGGCCATGAAGCAAACCAATGTAAACACTGGCTTCTCCAGAGATATC
AAAATCATCAATTCTTGCCCTACTGATGATCAGGAAGATGTAGAGTCTGATGAAGAA
GATGAATTATTCACATTCTCTCGTCCTGTCAAAGTTGAAGTGATTAACCAGAGCAGG
AAACCTGATAAGATTGTCAAGATGGTTCCTTCTGTCACTGTAGACCTTGAGAAATTG
ACTTCTCAATACCTCCTGGAGGATGAGTGCAATTTGGTTCTAAAGCTTCCCAGGGC
TGCAGCTGCCCAATCGGATCGCACC SEQ ID NO:259 Middle LPZ-233
GGTGCGATCCAGCTAATCAAACTTAATGGAGAGCCCTCCCAGGAAGAGTAA- ATGG
TAGTCACTTGAAGCCCTACACGGGTGGGCTGGCGGTCTGACTAACTGACC- AAAAC
ATAGTCTTCGCGACCCAACAAGCCAGACAGAGGTGTGGGACTATAAGCA- CAAGTAC
TAGAAGCTAGCATCAAAGTAGAGAATTAAGTTAGATACAGATGATTC- AGAAGCA
ATGGAGCAGATCCAGACCACGGTAGCATGGTGAGTTACGAACCTTCA- CGCCACAC
CAACGCAATTGGTTAAGACTTCGCACTAGGATCGCACC SEQ ID NO:260 Late LPZ-234
GGTGCATCCATAGTTCCTTTGCTAAGCGACTACTCTAT- CTCTTTTGACATTCTCC
AAATATTGGGTCTTTCAGTTCCTTCAAATGCTAGAAT- CATATCAACATGGGATTTAG
TGAGGCCGCAATACTAACCAGGGCATTAAAATAA- TACATTTCATTGATCCTATTCCC
AAAACATTTCCCGCTATCGTACGTTGACTCA- GCATATTTAGAGCAATTCTCTTACA
AACCTTAAGAAGGTTGTTCATGATAGTCT- TTCCGTCTGCAATATTGGATCGCACC SEQ ID
NO:261 Late LPZ-235
GGTGCGATCCCACCCAAGAGTTAAATTCACTTCTCCGCCTTTCTGAGGAAGAGCAC
TCTTTGGATGATATGAAAAGTGGTCCACTCTTAACCGTATTCGGAACCCTGTTC
CGCGGACGGTCGTATGGCGTAACCGGCGCAGACATTTTATCTCCTCACACAATATC
AACATTCAAGTCCCCGCTGTTCCCCGTTGCCTTTCTCTGCTCCCGACCGTTAAACA
AGAACGACCACAAGAATGAACAACACCGCAACCGAAACCTGACCCTCCACGTTGTC
TTCGGTTCGGATCGCACC SEQ ID NO:262 Late LPZ-237
GCGGACGCCTGGCAAAAACAGAGGGTATGCTCAAGCCTTACAGAAATTGAAAAATA
AGAGAACGTATGACCATCAATCTCAATCTCAAGAAAAGAAGTTGCAATACGACTCCA
ACACTTTTGAAAGTTGGAGGTTTGCTCTTTCTAGCGTTGCAGACATGGTTGGTTTTG
AGCTGGAAGCGTGTAACGGGCACTTTACAGTTGCGGGAATTGGAGATTGAGGACC
CCCTCTCAAACGTCGATAGGGAGGCTAAGCATCTATAGAGGATTGTGATTGGTCCT
TTTCCGCTACATGGAAAGTTTGTCAAACTCAGAAAATTACCAGAAGAATTCTGTCGT
CTTCTCGCAGCCGT SEQ ID NO:263 Late LPZ-239
GACGTTGTAAAACGACGGCCAGTGTAAAGAGCAGCCCCGATGCGCCGAAGCTCGC
GAGGGAAAAGCTGCAGAAGATGGGACCGATGACCAAGAATGAGATCATCATGAGC
GGCACGCTACTGGTCACGGTGGGTCTTTGGATATTTGGGGGTAATGCTGAACGTGG
ATGCTGTTACTGCAGCGATCCTTGGTTTGTCTGTCCTACTCTGCACAGGCGTCCGC SEQ ID
NO:264 Late LPZ-240 TACGGCTGCGAGAAGACGACAGAAGCAGAACCTGCCAATATAG-
GATCAATTGAATG TTGTGGGATTGCTGCATGCCCACCTTTCCCAGTTATTACTG-
CCTTGAAGAACCCAC AGCCAGCGAGTAAGGGCCCGGGTTTCGAACCAATCACAG-
ATGTAGGATAATCGCT TGAAACATGCATAGCGAATATGCCTTCCACATTTTCCA-
GTGCTCCCTCCTCTATCAT TCTTTTTGATCCTGCACCTGATTCCTCTGCAGGCT-
GGAAGAGTAATATGACAGTTCC CTGTAACAAATGCTGACGTTGTTGCAAAATCT-
TTGCACCACCAAGAAGCATGGTAA CATGTGCATCATGTCCACAGGCGTCCGC SEQ ID NO:265
Middle LPZ-241 TACGGCTGCGAGAAGACGACAGAAAAGAGG-
CAAACCGAGCTCGACACCTCCACTC AGAGCATTTGCAAAAATCCACAACAAATC-
TGGAGCCAAGGTCTTTCCCTCATTGAAA ACATTTATCGGACACATCAATGTCTG-
TAGTCTTTCCCATGGTCCATCCAGAGTAATC ACGGGAAGAACAATGCACTTCAG-
TCAGAATTTTTGATGACAGCTATCAGCTCCTG
ATCCTTTGAACCAGGTATATAATAATCTTGACCTGACTCCTGTTTCAACAGTGTAGA
GGTTCTGTCAACCTCAAGCAATGAATCGGCAGAACTTCCATTTGCTGTTTTGTCAAT
ACAGGCATTGTTTTTACCAAGACTGTGACGCATCTTCTGTCCTTGTCTATACAGTGC
AGTTTGTTCAAGCATAGACTTATGTGCTAGAACATGTCTTCCTTTTAAATTGTAAGA
GAAATGTAGGGGTTGACTGCTTTTACTGAGGCGTCCGC SEQ ID NO:266 Middle LPZ-242
ACGGCTGCAGAAGACGACAGAACCCTGGCTGACTACAACATTCAAAAG- GAGTCTAC
CCTGCATCTGGTGCTCCGTCTAAGAGGAGGCATGCAGATTTTTGTT- AAAACCCTTA
CAGGCAAAACAATTACTCTGGAAGTGGAAAGCTCGGACACTATT- GACAATGTAAAA
GCTAAGATCCAGGACAAGGAGGGAATCCCACCTGACCAGCAG- AGGTTGATCTTTG
CCGGAAAGCAGCTAGAAGATGGTCGTACTCTGGCCGATTAC- AACATTCAGAAGGA
GTCGACCCTTCACCTGGTGCTCCGTCTCCGTGGTGGCTTT- TAGGTTGGCTGTTGT
GTGTCAATGTAGTCTGGTGATGTTCAGTGGTTTTCCTGC- TTAATCCTTTTTATGTAT
GCATGTGTTTGTTGTGTTTGTGTTTTGTCTCTATGT- TTTTTCTACTTGGTTTGTCGGT
CGGTTGAAGCCCGGCTGGTGTCCTGGTAGGCG- TCCGC SEQ ID NO:267 Middle
LPZ-243 GCGGAGGCCTGGACAAACACAGA- AGGCGAAGTAAAAGCCAGTCTTACTTTTCATGT
AAATACTATCAAACTGCATGGCCGTTCCGCTGGTTGGCAATACCACACCTGCGCCG
GTAGTGCCAATGAACACTGCACCGGCAGCTCTTTCAGAAGTTGCAGAGGACTTACC
ATTTTAATTTTCACGGCATCCCGTCAAACGGCGGGATGCTTTTAATTTTTTAATCAA
AAAAAATATTAATTATGGCACACAATATTGTTTTCAACGAACAGACAGGCAAACACA
GTTTCTTTAGTGTAAAAGAAAAAGCATGGCATGGTTTGGGGCAAATTGTACAGGAC
TATCCCAACAGTAAAGAAGCATTGCAATTTGCAGGGCTTGATTTTGAAGTTTGCAAA
AGGCCCAATATTCACAGGCTTGATAATGGTAATGAGATTATTTCTACCAGTTCATTC
TATACTTACCGTCCTGATACCAACGCCATATTAGGCGTCCGC SEQ ID NO:268 Late
LPZ-244 GCGGACGCCTGAACATAGGAGCATTCTTAAGCATATCAGGTA- TAACCATAAACCTG
ACTTTGCTGCCCCGAATAAAGACATGCTCCAATTGGGATA- CTTTTCCATCCTTGGC
GTGTNTGTGATGCCCTCGAGCTGGCAATTCCAGTTATC- TTCGCATTCGATCATGCT
ACCCCTGTACAGCTCGCCACTTTTGAGTTCAACTGT- CACAACATGCCCGGCTGCTT
CATGGAGCAACTTCACAGGAATCCCCAAACTTCT- GCTCATTTTTTTGTCACTGCTCA
AAAACCCTAAACCCCAGATAAAACCCTCGGT- TCTGTGCCTTTTATCCCCGGGTGGC
TTATTGTTGCAGTAGTTGGCAACGGCTAG- ACTTACTCACATTTTGATTTCAATCTTT
CTAAGTTTGCCCTTTTGGGTTTTCCT- CACAGTAGATCCTATTTTATGTATTTTCTCGT
CTTCTCGGCAGCCGTA SEQ ID NO:269 Late LPZ-246
GCGGACGCCTGCAGGAATCGGCCGATTTGCAGTTCG- AGGCATAAGCGCATCGAG
GTCGCGTTCGATGTAGCAATTAAGCGCGCATGAACC- GCCGCTAAGCAAGCCAGTC
CCAATCAAAGCACATGCAAAGCGGATGCAATCAAA- TCTTCCGTTGTAAGCAAGCAC
AAATCCAACTGCACATGAGATCACCACCATGAA- TGCAATTCGAGTGCGAGCTAAAT
CCCAAAACGCTGCGAGTGTCCCCTGAAGGCG- ATTCGTATGTAATATTTGACCGCTG
CTCAACACAAGCAGTACTCCAAACACCAG- TGCTTCCGCCGTCAATTCTGTCGTCTT
CTCGCAGCCGTA SEQ ID NO:270 Late LPZ-247
CTGCGAGAAGACGACAGAACACAGACACAAAATTTGGAAACTACA- GAAAAGACCAT
GTCATGAAATCTTCATAATTGGGCTTCAGATGCAGAGGGGGTC- GGTTTTGGATTAA
GCAATGGCTGAAGTGCTTTGACAACAATACTCATGTTAGGA- CGAAAATCTGCTTCAT
ACTGCACACACAATGCCGCAACAGCAGCCATCTTTGCA- ACAGCCTTTGGAGGATAT
TCACTCTTCAACTTGGGATCAACACACTGCTTTACT- TTGTCTTCACTCAATCTTGGA
GTTGCCCAAGTAACAAGGCTTTGTTGTCCCCTA- GGCATTGTATGGTCCACAGGCGT CCGC SEQ
ID NO:271 Late LPZ-248
TACGGCTGCGAGAAGACGACAGAAAGAGACAGGCTTGGACTTCGTGGCCTTCT- TC
CACCACGCATTATTTCTTTTCAGCAGCAATGTGATCGTTTCATGGTTTCTTT- TAGAT
CCCTGGAGCATAACACTCGAGATGGTTCAGCTGACTTAACAGCTCTGGC- AAAATGG
CGTATTCTTAACAGATTGCATGACAGAAATGAAACACTATACTACAA- GGTTCTTATA
GATCACATTGAAGAGTTTGCTCCAATAATCTACACTCCAACTGT- AGGATTGGTTTGT
CAGAATTATGGTGGGCTGTCAGGCGTCCGC SEQ ID NO:272 Early LPZ-249
GCGGACGCCTCAATAGTATGGAAGGGCAGCTGCACTACTC- AGCATGAGTGGAG
GCCTAAAAGTTTTGTTAATCTTTCTGGTGAGGTGGACACCA- AAGCCCTTCACAACA
GTGCAAAGGTGGGGCTATCTCTGGTTTTGAAGCCTTGAA- GGATATGCACTATTTGG
TACAGATTTAAGCGAAGGTCTGTGCCAAATTTTTATT- GGAATTTTTGAGTTTTTCCTT
TCAGAATAATTATTTCAATGCCTGTGTTTTCTG- TCGTCTCTCGCAGCCGTA SEQ ID NO:273
Late LPZ-250
GCGGACGCCTTTTGCCCAATTAACATCCCTGCATCTGCGCATTAAAAATTGATTGC
AGACCTGAGGTTTAAGTGGAAGCTTCTTCCACCATCTCTCCCCTGTTTAAGGAAGA
CCCGAAACCCTAGCCACTGTCTCCTCTGTGACTTAAAATTCCAGTTCACCAACCTTA
ACTCTGCGTCCGTTAAAATTCTGGGCAAACTGCACTGCCAATTGGTCATCATATCCT
CTGAATTTGGCAAAGAAAACATAGGTCATTCTGTCGTCTTCTCGCAGCCGTA SEQ ID NO:274
ND LPZ-251 GCGGACGCCTCGTCAATCCATGGTTGTAAACATGCCTTCAAAA-
CTGTTTCCTTATGT CGCACAATGTCTACATGTTCCTTGAGCGATTTTTCCTGCT-
GCATTGCGAGCCTCTG TGTAAGTCCCACTATCTGCGCTGTCCCTTTTACTTCAT-
AATACTTCTGTCGTCTTCT CGCAGCCGTA SEQ ID NO:275 Late LPZ-255
TACGGCTGCGAGAAGACGACAGAAAAAACTGTATACGAGTAGGCAGCGAGTCC- TG
GCAGTATGGGAGATTGAACTCCAATTACATTTAGTTACAAGTAGCATCAACA- GTGAC
TGAGCCAAGAGCTCTACACAGAAAAATAAAATAAAAACTGTATATATTT- ACAGGAGA
AACCCCTATGGCCTCAGGGCCTGAATAAATCAATCGCAGCGGTGGT- CGATGTGGC
CTTTTCAGGGCTGCAAATCTTGCAAGGGGAAGCCATCATCCTTGT- TCCGTATCCTT
TTTGAGGGATAGCGAGCCACGCAGCCAAGATTTGAAGCGATTG- AATACTTTGGGGT
GTCGAGAACGCACCAGAACAATGCCACTCGAGAAATACTAC- TGTGATTACTGTGAC
AAACAATTCCAGGATACTCCCTCCGCTAGAAAGCGACAT- CTACAAGGCGTCCGC SEQ ID
NO:276 Late LPZ-256
GCGGACGCCTGTACCGTATTGGAATTCTAAACCCTTCCTTGGTATAGGGTTTTCGC
CACCCTTGCGTTCATTTGGTTTTGTATTACGTCCGATTCCTCCGTCTGCGAGCTCTC
TGCAACTTGGCAATTTCATTGTGATTTTATCCTATGATGCTTCGTATTTGTTTGAAGC
TCGTCCTCCTAGTTCTCTGTGATACCAGTTGGTAGTCTGCAAGTTTCGATGTGGGT
TCTTTTAGCTGGTCTGGGGTTTTGTTGCTCTGAGTATGTTGAGCTGCATGCTCGTG
GCGGTCTTCACGGCTCCATTTGTTCGGAATCTGTTGTGGAAGTGTCTCGGTCATCT
GTGGAACTGTGGAAACCTGGTAAGATTTGTTTATCTGCTTGTGTCTAAACTGTTCTT
GAGTTTTCTGTCGTCTTCTCGCAGCCGTA SEQ ID NO:277 Late LPZ-257
GCGGACGCCTGCTGTTGAAGAAGGATGAAGTCATTGTCTGCGGCCCTGTTCAG- CA
TGATTTCGGCATTCTTAATCTGGTCAACCAGTCAGAAGGTGGCGCTGAAGGT- GACG
AAGAGGCAACCTGGGTAGCTGCACTGGAAACTCAAGCTGCAAGGGGCACC- GACCC
TCAGACTTCGCGCGATAACTTCTCCCTCTGGGTAAGTCGATGCCAAGGT- CCTTGT
TCTGGGTTCTTCTCTCTGTTTCGCATGTTGTTCTTCTCTCTGTTTCAT- TTGTTTTTCT
TCTGTCGTCTCTCGC SEQ ID NO:278 Late LPZ-258
GCGGACGCCTGCACATACAAAGAACGACAAAAACAAAAGCATAAAATCCAATA- GAT
GCAACTATATATCAAGTCAGAAATGATATAACTCATCATTATTACAAAGAA- CAATAAG
AGTGGAACCATAATAATAGTCGTCTATTATTGATAAATAAAGAAGAA- TACAACCATA
GTTCTGTCGTCTTCTCGCAGCCGTA SEQ ID NO:279 Late LPZ-260
GCGGACGCCTGTATAACATGCACCAAGAGACCCAATCAAAGCACAT- GCAATCTGTA
TATATAGCAGAATAACAGCCAGGGATTGCACTCTATCGTAATCG- CGAAACCACGCA
CTAATATGTGCCCATGCTGATGATGCACACAGCATGTTCTGT- CGTCTTCTCGCAGC CGTA SEQ
ID NO:280 Late LPZ-261
GCGGACGCCTGAACTGTATAGAGTTGAAACTTGAGGGAAGGCTTGCTGCCACCAA
AGCCTCCCTCCTCTTTCCTTGGCGGTTCGTCACCTCCTTTCGCGTCAGAGCCCCAA
TTCCCCTCCTGCGCACACCAGCAAACTGCATCGAATGTTTTTTCCACCATTCTGTAA
ATTCCCTCGGAGTTACCTTGGGGCAGAAGCCGCATTGAAGAGCATTGAATGCTATT
CATTATCCCACCGTAAACTACCATTGCAACCTGCCTGTGTATCGACCCGCTGTCCT
CTACGCGTGGCTGGCACATGGCGTCGTTAATTGCATGTTGACACCCGTATCCGG- G
TGTGCTTGTGTGCTCGTCTGCATATCATGTTTTAGGATCTCATAGAAGGTGGA- CCA
TTCTGTCGTCTTCT SEQ ID NO:281 Late LPZ-264
GCGGACGCCTCTTACAATGTCTCTAAAGATTGGAAAGATTGTCTTGTCTGCAACC
ATAACTTCCGCGTGCTTTCTTATTAATGCAACCCACTGTGATCCTTTCCGCCATTTA
TCCTTTCGAATGGTTGGAGCCATTTTTGGGTTGTACCGACTAGCTTTTGGGTCTAC
AAAGCTGTCTACAAAACTCTTTGGAGATGACATTACATAATCATATGTATAGCTGAA
GTTGTACAAAGGTACACAACTATCTGAAACCAAAATGAATCTCTCGTTAGCTGG- ATC
CTCGAGTGCTTTCCTAAGTAGAATACGCTCCGCTTCTATCATACTGGCTTC- TCCCC
AAGTACCTGTATGCTATCACTAAGCTGCCAGCCGTAACAAAATGTACAT- TCTGTCGT
CTTCTCGCAGCCGTA SEQ ID NO:282 E,M LPZ-265
GCGGACGCCTTGCTAGGAGAGCTCTACGCCATTATTTGAACGATTGAGCCGAAGTT
TCACCGTTTAAGGCATTTGTGTCCCAGAGGTTATTGGAGATTAGCAGCTTGGATTT
GGCTGCTTCGCTCAGCGCCGTGATTCAGCTTTTGATTGATTCTCTCCAGTTTCAT
CCTGTAACGACAATGGCAATGAAGACCTACACATTGCAGTGGCAGCTGCGTACGC
TGTAGTCCTGATGTTCGCTCTCTTTGGCATCGCAAAGGCTGCTGATGCACCGTC- TC
CCAGCCCCGTTACTGGCGCGGGTTCCATGGACTTCGTTCCTTCTGTCGTCTT- CTCG CAGCCGTA
SEQ ID NO:283 Middle LPZ-266
GCGGACGCCTTATCAGCTGGGGGCATTCATAGGTATGGAAATTCAGATCAACTTCA
GTGGACAGTATGTGGATTTAGGCGACCTGTGACAGTTCACGATATCTATTCATTTCT
ATCCAGAGACAGATTCCCATACTCACCTCCGTCCTTCCCATATATTTTCTGGAAGGC
ATCATGTCCTCCCAAATTTACTCATTTTGCCTGGCCGTCGTTTTACAA SEQ ID NO:284 Late
LPZ-268 GCGGACGCCTGTTGCCACAGAAGAATGAATAATGCTTCA- AATTTGAGACCTCTTC
GGAGGAAAATCCTTGTTCTTACTGCCTAACCACTCATG- ATGATCTGCGTCACGCTG
ATTATGAGCTGCAATTTAAATTATTTCAGATGAAAC- ATTCCCATATTGAGCTTGCAG
CAAGTTGCAGACCCTTCAATTTCAGTTCTGTCG- TCTTCTCGCAGCCGTA SEQ ID NO:285
Middle LPZ-269
GACGTTGTAAAACGACGGCCAGGATTAAGGTTCATGAGCTCCGCAACAAGAGC TCAG SEQ ID
NO:286 Late LPZ-270 GCGGACGCCTCTAGGAGCCGGCGGA-
ATTCCTGTGAGCTCGAATTTGCCGAGCAG GTTATTGTCCTTCGTCCGCGCTCGC-
TCACCTTCATATACTTGAATTAGAACCCCAG GCTGATTATCTGAGTAAGTTGAG-
AAAATCTGCTCCTTCTTGGTTGGAATGGTGGTG
TTCCTCGGTATTAATACTGTCATTACACCTCCCGCTGTCTCCAACCCCAGACTTAAT
GGCGTGACATCTAGCAACAGCAGGTCCTGCACCTCTCGTTGCCTTCGCCGCTGA
GAATGGCAGCCTGCACAGCTGCACCATATGCCACGGCTTCGTCTGGGTAATGCT
CTTACAAAGCTCTTTGCCATTGAAGAAATCTTGGAGCAATTGTTGTACTTTGGGGAT
ACGAGTCGAACCCCCGACCAAGACGACATCATCTATTTGGCTCTTGTCCATCTTAG
CATCTTCGCATACATTTCTCCACAGGCTCCATACTTCTCCTGAAAAGATCCATGTTG
AGTTCCTCGAAGCGAGCTCGCGTAATTTGTGGCGTAAAAATCAATTCCTTCATATAG
AGAATCAATCTCAATCGTTGTCTGTGTAGTAGAAGACAGCGTTCTTTTGCCCTCTC
ACATGCTGTTCTCAGCCTGCGAAGAGCTCTGGCATTCCCGCTGATGTCTTTTCTGT
GCTTTCTTTTGAATTCCTGCACAAAGTGATTCACCATTCTGTCGTCTTCTCGCA- GCC GTA SE
Q ID NO:287 Late LPZ-271
TAGCCATCGCCATTCTATAATCTTAGGATCCTTGCTGAACGATAAGCCCATAAAAT
TGATGCACTGCCTCGCTATCCCTGGCCGTCGTTTTACAACGTC SEQ ID NO:288 Middle
LPZ-272 GACGTTGTAAAACGACGGCCAGGAAATTACAGCTACCTCTAACTGGTTTGA- CGGCG
TTGCATCTATGAGCCGCAAGGGTTCGAATCCTCTGCGGGCCAGATCTGC- GATGG
AACCCTGGGCGAGTGCAATGATGATGAAGAAGAGTTTGCGATGGATTCT- GAAGCG
CACGGGAGGCTTCTGAGGAGGATCCGTTACTATATCAGCTACGGAGCA- TTGGCTG
CTAATCGCGTTCCTTGCCGACCTCGGTCTGGGAGGTCTTATTACACT- CGGAATTGT
TACGGCGCAACAGGCCCCGTCAGACCTTACCACAGAAGCTGCACT- GCTATCACTC
GTTGCAGGCGTCCGC SEQ I D NO:289 Middle LPZ-273
GCGGACGCCTGGGAAGCAATGGATGGGTGGCTAGACGCCATCCGTCTGTGT- ATA
CTATTTTTGCACGCGGAAAGAGTGATGTCCTGGCCGTCGTTTTACAACGTC SEQ ID NO:290
Late LPZ-274 GACGTTGTAAAACGACGGCCAGATTCAAAA-
GAAAAAATCCTCACTTCTTGGCTCCG TTTGCGCTCCCGCCGAAGCTCCTCTGCA-
ACCCCTCTGCAGCGTACACTGCATCCC GCTCGCGGTGCTGGCTCACCTCGCAGG-
TCCGCTGACGGTAAATGGTTTCCAATAA AGCTATTTGTCCTCTACCCAAAATCC-
ATCTAGCATTCGTTGTGGATTGACATTCTGC CATTTCTCTGCTTTTCTGGTTGA-
TATGCAAAGATTGAAAGCCCAATTGCAAGCAGTG GTCGTGGATTCACTATAAGGCGTCCGC SEQ
ID NO:291 Late LPZ-275
GACGTTGTAAACGACGGCCAGGAATAAAACAAAGCATCACTGCAAAATTTCAAAC
GTGGTAATAACGGCTAGCCAGCTCGACGTGAAGGCAGTGGGGGCCTTGAGGTTGC
CTTTTGGCGTTCAAAATTGGCTAGACTACCATAACATAAATATTGATTTCTCAGTGA
CATCACTGGTTTGGAGTCATCCACAGCCTGTGCACCAGTACGGCAATTGCCTTTTA
CATGAAGCCATCCTTTCACTTTTACTTTTGAGATTCTCAGAACTGAGGGGCTAGGC GTCCGC SEQ
ID NO:292 Middle LPZ-276
GACGTTGTAAAACGACGGCCAGCACCTTCCTAGTCCCCTGTCCATTCTCCTGAAA
TAGGAGCAGTTTGACCCAGTCCAGTTTTCAGAATTGAGAATATGAAACAAAGAACC
AAGCATATGAGAGAACATACAAAGACTTTGTATAAACTACTTTTCACAGGATCTCAA
CAGCCCTCTGCTGAGATCCATTTGATACAAGGCCCCTTGCATCTCCACCCTCTCCC
TTATCACCTCCACTAGAAAGATGATGGAAAGCAGACACATGGAAATGTTGCTGCAG GCGTCCGC
SEQ ID NO:293 Middle LPZ-277
GACGTTGTAAAACGACGGCCAGTTAGGTTGTATATTGATTGATGACTCTTTGACTCC
ATTTATGAAAACATCTTTGTTCTCGAGATTTAATCAGTATTAAGCTTTCAGAGTGAAG
TTCAGTTTGATCTGCATAAACCTGATCCACCATATCTACATCACATCTAAAATTACTA
AAATGTGAGGAGATGGAATTTGTTTCTTGAGAATCCCTATTCCTCATCGACACTGTT
TACTGGATCAGATCCAATCAAACTCTTGAGAAGTAATCTCTGGAAAGAAATTAAAAA
GTCTTTACCTGAATTATCTCGATATCAGAAGCAGAAATTATGATACATAGACTTCT- TA
ATAATGAAGAGTCATTTTGCCAACGTTGTCTTTGCCACCCCACCAATCCCCA- TGATC
CCAAAGATCTGAGGTTTCCATCTCTATGTGGCTGTGATAACACTGGATT- TTTCAAAA
ATCTTCTACTTTCGCATCCAAACCTTTTTGGGATATTT
SEQ ID NO:294 Late LPZ-278 GACGTTGTAAAACGACGGCCAGGGGGATGGGAGATACA-
GAAAGATTCCGGATAAA AGGGAGCAATGAACGGCTGGTTAAAGCGTAGTCCACC-
ACACTAGCCCCACCTCCA TGAGGCCTACACGTGAAGAAGCAGGATCTGGGAAGC-
GCGAGAGGCCGTCAAGA TTATCAGCTCATGTGATTCGCCCAACTGCAAAAGATG-
TCTACCGTAGGCTGTGATG GGGCCCAAGGCGTCCGC SEQ ID NO:295 Late LPZ-279
GCGGACGCCTATCAGATGGGTGAGTTGACCGACATTTATCGTCCGA- TAAATGTTTG
AGGCTGATGTCATGGCAATCCACGTGTCTGCACCATATTTCATC- GGAGCCCCTCGT
CGGAATATTCCATCGCCGGAGAGCTGGCGCGATAGGTTTCAG- GCGGCCGGTTTCT
GGTTTGCAGCTGTGGCTTCCCGCGCGCCTTAACTGTTGGCC- CGCGCGCACAGGG
GAAATTACAAATTTCAACATATCCAATACCATCATATAACC- CAACAACACTAGCAACA
GATCCTGTTCTGTGCCATCGTCCAACTCTGA SEQ ID NO:296 Late LPZ-280
GCGGACGCCTTAATTCGACTACAAAGATACTGAAGC- CAATGATGACAGGTTGTGCC
ACTTTCCCAGCTGATAAAGACAGCTCTGAAATTG- ATAGAGCCAGAACTCCAGCTGC
AATGCTCCCCAGAGCCTGGTTGAAGCGCTTGC- TAAAGGTGGCACTTTATAGACCGA
CCCAAAACCTCCCTGGCCGTCGTTTTACAA- CGTC SEQ ID NO:297 Early LPZ-281
GCGGACGCCTACTGGAAACCCGGTC- CACCGAAGGCTGAAATTGTCCTGCTTTGTA
TACCGAATGGCAGGAAGGTTGTCG- AGCATCAGGTTCACCTGGTAAAGATTATCGAT
CCTATGCTTCAATACCTTCAGCTGCTCTGCCCCAAGGACAGTAGTATTGCACAGGT
AAATTCAGATTCATTGACATTCATCCGGAAGCGATATGGTGAGTTCTCGATCCTGT
CCCCCATGAGGAGCTCCCCAAGATTTTCTGCCATGTCCTTCACACCATCCAAGGGC
TTGCAGAAGGGCAGGCTGTAATAGCTGTAGGGAAGCTCTGTCTCGACTGAGGTAA
GGGAATTGACGTTCACCCATAAATCTGACCCCTGGGAGAATATGATGTGAGGAATA
CAGTGCCCAGTAAATATAACTCCGCATTATACGTTTGTGTGTGCCTTCCCCAATATT
GCCCCAACATAATCAAAACCCACAATCCCAAATCCTGGACCGTCGTTTTTACAACTG TC SEQ ID
NO:298 Early LPZ-282
GCGGACGCCTTGTCAGGACCAAATGTGTAAGAAACACCTCTGTCATTCGAGCCCC
TCCTTGAATTGCATTGCAGGGGTCTGACCAAAGAAGATCACATTAACAACCCTGTAT
CTGGCACATCTGTAGGTCGAGGTATATTCTTTATTTGTTCCAAATTGGTCAGTTCAG
GCGAAAGACCACCATGCATGCATAGGATCTTTTCATCTATAAGTGCAGCAACAGGC
AGGCAGTTGAAACAGTCTGTAAAAAGTTTCCATAGTCTTACATTGAATCTGCGCTTG
CACTCATCATAGAAACCATATATGCGATTTATTGAGGCACATTCATGATTTCCCCTC
AGAAGGAAAAAGTTCTCTGGGTATTTAATTTTGTAAGCAAGGAGGAGGCATATTGT
CTCTAGGCTTTGTTTGCCCCGGTCCACATAATCTCCCAAGAAATAAGTAATTTG- ATT
CTGGTGGGAAGCCACCATATTCAAAAAGCCTTAGACAGATCAGAATACCGG- CCTGT
CGTTTTACAACGTC SEQ ID NO:299 Early LPZ-283
GACGTTGTAAAACGACGGCCAGGAGACGGGAATACCTATTTTTGGGAGGATTATTG
GGCTCGGGAATCAGCATATTGATGTGGCTGCAACTCGCATCCTCGATCTTTGGTGG
TTCTTCGGCGATTTACACATTTGAGATCTACTTCGGTCTGCTAGTTTTCCTTGG- GTA
TATTATATTTGACACACAGATGATCATCGAGAAAGCGGACCATGGAGACTA- TGATTA
TTTAAAACATTCACTGGACCTCTTTATTGACTTCGTTGCTGTATTTGT- TCGCCTGAT
GGTCATAATGGCAAAGAATGCAGACAGTAAATCCAGGGAAGGGAA- AAAGAAGAGA
AGGGCTTGAACTATGTGAGATACAAAAATATCGAGAATAGAAGG- GCTTGAACTAGG
GCTTGAAAGCGTCCGC SEQ ID NO:300 Middle LPZ-284
GCGGACGCCTATCAGACAAGGGTTGTTGACCGAACTTTATCCTCTGAAAAG- TGCTT
GAAGCTGATGTCATGGCAATCCACGTGTCTGCACCATATTTCATCGGAG- CCCCTCA
CACGGAAACAACCTTAAGCCAAAAGGTGGTGCGATGACTTACCGGCC- GTTTATGGT
TTGCTTCGGTGGTTTTCTGTTGGGTGGTTTCCCGCGCGCGTTAAC- TGCTGGCCGT
CGTTTTACAACGTC SEQ ID NO:301 Late LPZ-286
GACGTTGTAAAACGACGGCCAAGAGGGGGAAACTCCCAAAACACTTTTCCATTTTT
CTTCTTTTATTAAACTTCAAAGTATTTTCCAACAGAGTTACAAGGGGCCAACCATGT
CCAAATCCATGCATTTACCAAGTACAAAGAATGGTAGTCCTTGGCTTGACCTAT- CG
ACTAGCCAAAAGTGCCAAGTCCACAACTAGGGTGTGCCCAACCTAAGGTGAC- ACC
TTGCCTAGAAAAAACCCCAAACTTGGCACCACAAATAACACAGAAACACAA- CTCTTG
ACCTCTGCCAGAAACCAGGCTCTCTTGGGAAAGCCACACCTCTCTCTG- TGATATGT
CTTATCTCCAATTTCCCTTTTTGTGATGCACTCCCTTGCTTGTGGT- TCTGCGATATC
ACACAAACTTACATTTCTGCGATTTTTGTTTCTTGCTTCTCCA- AATCATGCGATCTTA
TTTTTAACCCTTGAGACCCTTCACACTTTCCATCCATGA- CGTCACTTCATCGTTTTA
GCCAATTCGTCATTTGGGCATGTTGGGCGTTGGGTC- TACCCGTATTCCGGTCGTAC
AGGCCAAATTGACCATTTTGGTCCAGGTGGGTGC- ACCCATTCCTGGAGGGCGTTC GGC SEQ
ID NO:302 Late LPZ-287
GCGGACGCCTCCACAGAGCTCACACATACAATATACTATGATGCCTCCAGAACTAT
GGCACTCTGTATGCCGCTTCAATATGGATTAGCCCACACTGCGCCATCCAATTAGG
CGAATCAACCTTATAGCACCATCCACAACCTCCAGCGCTCTCTTTTTCACGCTAGA- T
TGGCCAACTACAGGCTTTACAACACTACTCATATACAACTCAACTCGGCTCCT- CTGC
TCACCACTAAATCACACAGGCTCCAATCGCTAGACAGAGCCACTACACAG- GCACTA
ATAGCCACTACACAGGCACTAATCTTGGCGTCCTCCACCAGGTTCCAA- CAACAACC
CCAAATTGCATATGCACTCCACAGTGAGCACCAACTAGGTCCACAC- AATAGGCCAC
ACCAACAACACTCCAAGGACCCTAGATCCTGCCTCACCCAGACA- CCACTAGGCCTT
CCTCACAGCTCACCTAAGTGAGCCAACAACTGGCTGGGCACA- CAGCTCCCAACTAT
ATGAGCACACAGCCCAACTACAGCTCCAGCACACGCACAG- CTACACGCACAATGC
CTTCTCAAGTTCACAGCCACACCATAACGCAGCACAGTT- CTTACAAACATATCTCTC
CAGGCGTCCGC SEQ ID NO:303 Middle LPZ-288
GACGTTGTAAAACGACGGCCAGGATAATGGACACGAGAAACCTTTGGATGT- GCCT
CTAAAGTGCGGGCAATCCTTAAAGCTGTTGAATTTGTGCTGTACACGAAG- GTGC
AGGGTCTTTATGCCACGAAGAATCAAGTACGCTGCATTTGGACTTAATAC- ACCTCC
CAAGACATTGTGCAAAGCACGTACTGTGCCAATAACCTTGTTTGAACC- ACTCAAACT
GCCTGCAAGAACATCATTATGACCTGCAATATATTTAGTTACCGA- ATGCAATACAAT
ATCTGCGCCGAGTGCTAACGCTTTCTGGTTAACAGGCGTCCG- C SEQ ID NO:304 Middle
LPZ-289 GACGTTGTAAAACGACGGCCAGTCATT- ATTGACAATAATCCTTTCAGCTTTTTACTG
CAACCTTTAAACGGTATACCTTGC- GTTTCTTTCACTGGAGCACACTCAGATGATAAT
CAGCTTTTACAGGTGCTCTTACCTCTGTTGAAGCATCTTGCCACTCAGGAGGACGT
GCGCCCTGTGTTGTATGAAAGATTTTACATGCCCGCATGGTTTGAAAAGCGTGGCA
TTCCAGCATCTGAGTGGCCCTTGTGACTTGGTTTTGATTTTGGATACTCTTTGTCAT
TTTGGGTCAAGGTAAAGGTGTACGTATCCAAGTGATGCAAGCGTCCGC SEQ ID NO:305
Middle LPZ-290 GCGGACGCCTGATAGCACGAGTCTTCTGGGACGCAAATCAAGAG-
GCAGGTACTT CTTTTTCTTGTATGCTTCTCTTAATGCGGATCGCTGGCTCTGAG-
AAATCACAGTCAG AACCTGAGCTATTGATAGCCTCACGACCTTGATTTTAGAGA-
GTTTGTTGGGCGCTC CTCCAGTGACCTTGCAACTCTGAGCAAGGCAAGCTCAGC-
CTGAGCTCCTTGACC TGGCTTAACAGCTCGGATTTGCCCTTGTGGCGGACTCAA-
GGACCTTTAACCTGGG CGTTCGT SEQ ID NO:306 Late LPZ-293
GCGGACGCCTGGTGTCGCTGGGCCAGTTCAAGTATTTTAGCAACAGTGTTCACACT
TATTCCCTGTGATATTCTTGACTCACACAACCACCTAACTGACGCAGACCATATCG
ATCTGCTGCTGTAAGCAAATGTTCGATCATTGTCTCAGGTGTCAAAAAGCAAGGGG
ATGGATCAGAAAGCTCTTCTAAATCTGCATGCTCCTCTAAATCTGGAAGGGTAT- CTT
TGTAAATAAAGTGTAACATAGCCTTAAACACCTCTGGCCGTCGTT SEQ ID NO:307 Late
LPZ-294 GACGTTGTAAAACGACGGCCAGAGGTGTTTAAGGCT- ATGTTACACTTTATTTACAAA
GATACCCTTCCAGATTTAAAGGAGCATGCAAAT- TTAAGAAAAACTTTCCTGATTCAA
CCCCCTGCCTTTTGGCACCCTGAAGATGGT- TCAACAATTTGCTAACGGAACCAATT
CAAAAGGGCCGCCTCCATTTAAGGTGTT- GTGTTAGTCCAGAATATCACAAGGAATA
AGTGTTAACACCGGTGCCAAAATACC- TGAACTGGACCAACGACACCAAGCGTTCGC C SEQ ID
NO:308 Middle LPZ-295
GCGGACGCCTTGTAATCCAGGGCCTTGAATATTGTAAGAGAAGATCGAGAA- ATAAT
AGTTTTCTTATTATCAGGAATCACAGCTTGAAGAAGGCAGACCATGGAC- TCCCACT
GGCTTCGTGATATTGAGTCCCCAACAAACATTAGTCGTTTTCCCCTC- AATCTCCACA
GCAAGTCTCTGGCATTGAATCTGCGAAAGGAACACCCGAGTGGC- TTCCACCTCCAT
TTCTCGTAATCAGAATCTGGCCGTCGTTTAACAA SEQ ID NO:309 Late LPZ-297
GACGTTGTAAAACGACGGCCAGCAGAAGACCAGTGCAG- TATGCTGCAGCATAGTT
TGTAAGCCCTACTTCGAGTCCATAACGAGGCAACTCC- CTAGAATAAGCAGCCGACA
TAACAACATCTCCCGCAAGAGTTGCATAAATGATC- TGTGCCACCACATCCTTGTTG
TGAATCTAACGACCAATCGGTATTTGGGTGTGT- TGTACTTGTTCTTATCTTGGTTAA
TCAGGCGTCCGC SEQ ID NO:310 Late LPZ-299
GACGTTGTAAAACGACGGCCAGCATCCATTGCAGAAATTTTGGGGG- CTATATTTAG
CAACAGATATCACAGCTGTAAGTCAAAGTTGGACCCTTCTTCTT- CGACATCTTTTC
CAGCTGTGCAATAAACTGAACACTGTCCTTTTGGATAAGCTC- CTCAACATATTTAG
AAAGTTCAACATCCAAGACATTGCGGTACTCCTCAACATA- TATGGATGCAAGTTCAT
CATCTGCAGCTGGTCTCACCGCTGTACAAACTGTTTA- ACATGGTTGACAGTTGCA
AGTTGAGCAGTCCGTGGATCCAAATAATGAGTTCCG- TCAAGCTCACTGAACTCAGT
CACAATCACCTGGCCACTTTGATTGGGCATCTCG- AGGGATATCATGTGAGACTTGT
TGTGGATGGGGAAAGCGTCCGC SEQ ID NO:311 Early LPZ-300
GCGGACGCCTGCATAAACATCGCTACCCTGGGGATGAT- TAATAATAGTACCAGGGT
TAGGATTTTCTTCATCTTGAGCGATATCATCATACA- TAAAGACCACAATGTTTTCCTC
TTTCAAACCGCCTTTCCTCAGAATTTGGTAGG- CATGGCAGATATCAGCCTGATGCC
TGTAGTTCCAATAACCGGAAGAACCAGCCA- ACAGAATAGCCCACTGAGTACCGATC
GTATCACTATCATCAACGATATGATCGG- TGGGCATTTTCAGTACTGAATCCCAACCC
CTTCTGGCCGTCGTTTTACAACGTC SEQ ID NO:312 Middle LPZ-301
GCGGACGCCTAGACTGGGCATACCAACT- ACCTTCCTCATGCCAGGCCATGGGCCA
CCTACCTGGTACTTAGGCATAACACCT- TACTTACGAGCATGCCAGGCTCAGTCAGA
TAGGCATGCATCCCACCCACCTAGC- TATGACCCAATCCTTATAAACACTAGATATTC
TCCCTGGCCGTCGTT SEQ ID NO:313 Late LPZ-303
GCGGACGCCTAGACAATCATACTGAAGATCTGTAAGC- CATGACAAGACGAATAA
AACGAAGCACGGCGCAACCAGCGTGAATATTGACGCC- TTAATTTCATTCAACTGGG
TTGCGGATTCTTTATTCCTCAACAAGTGTTCGATA- GCTTCACATACGCAAGGCCCCT
TTTACTCTCACCTTCATGGTTTAATGCTGTAA- CCGTCGAAGGTTGATGAAAGGACTT
GGATGATGATGTTGCCAAAAAAAAAAAAA SEQ ID NO:314 Middle LPZ-304
GCGGACGCCTGCTCAACACCTGTATAGT- CATTTCTTGTTTCCTTTTCTCAATTTTC
TCTTTCGAATGACCGCATTGAAATTC- AGGCTGCCCAACGCGTTTTTGTTTTCACAAT
TAATTTTTGAATCATACGCGAAG- ATCATGATGAGAATGGTTGTGGAAAAAAACTGTT
TGTAAATATTTAG SEQ ID NO:315 Middle LPZ-306
ATATCACATTACCATTCAAAAAATAAACATTTTAC- AAAATACAATTCCATAACAATTTT
CTTCCCTGTTCCAACCTCCACAAAAGTAAA- TGATCGTATAAGAAATTAACTACCAAC
AAAAATCCCAAAGTTAAAGGAAGACAT- CCCCAAAAAAGATGTAACTTTCAAAACCGG
ATGACTTCACTCCTGCCATTGCAC- CTAGTCATTTACTTCTCAGAGGAGTTTGGCCCT
TTCTTCTTTCCAAAAGTAACCACTGCGGTAAGAAACCGGCGGTTGTATTGCATTCG
CTTGTAGGCGCGGCCTCTAGGCTTCTTCTTCTGTCTTGTTTGGCCACCTTAGGGTC CGC SEQ ID
NO:316 Middle LPZ-307
GCGGACGCCTTGGTACAATGGACTTGCAAAAATAAAATGAGTTCTCATTGTGGGT
GAGATGCGGATATTTTATGCATAGGCACTTCATGGAGATGTGGTTATAAACGCCA
TCTTAATATCTGTACCTATTACTTTCAAAATATGAAGGCAAGATGGAAAGCTACTCAT
CTGTTGTGAAGTCAGAATGTTGGTAGCGGTTGGGCTCTGAAAGTAAGAAACTTTTT
GATTGGTTTAATTAAATGAGGGAATTTGCCTGGTTTCCCTCTTCCTTCCGAAAAAAA ATTTATTTA
SEQ ID NO:317 Late LPZ-308
GACGTTGTAAAACGACGGCCAGACAATATTGGAAGGGAGAAAGGCGCCAGCAGGG
TTGAGGGGAAGAAATGCATAATGACATATATAATGAGATCTATTTGTATACGATATT
ACGGGTACGATCGATGATTCGAGCTACGATCCCATACGACGCTAAAGCGTAATTAC
ATATATAATAGATGCATTTCAGAATGACTTATCTATTTCATTACGCGATATTATATAC
GTAATTACGTATATAATTGCAGAGATCTCACCGACCAACCAAATAGTCTTTCATTTC
ATCCCAGGCGTCCGC SEQ ID NO:318 Late LPZ-309
GCGGACGCCTGTATCACTAGAGGTGAATACTCAGCAAGCAAAACTGAAGGATATTA
TTGAAAAAGCTGTCAAGGCTAAATTGGGTGTCAATTCCCCATTGATCATGCATGGTT
CTACACTTTTGTTTGAGTCCGGTGATGACATGAGGAAGATGTTGCTGCACATTAT
GCACAAAACTTAGAGAAGACGTTAGCAGAATTTCCAGTTCCAATCACAAATGGTGTT
ATTCTTACAGTAGAGGACTACCAGCAAGAGTTCTTATGCAGTATTAATATTAAGCAC
AGAGATGACTTTGATGAGGAGTCAGGTGGCATTGTACTGTCTGGAGGCGTCCGC SEQ ID
NO:319 Late LPZ-310 GCGGACGCCTCCTTGTAGATACGATACATGAGTCTA-
AGATCAAAATCATACAAGAA GAGCTTCATTTCCGGGCCTCACCTTTTCTACAAG-
CTCCTTTTTGGCTGGTGGAAAGC CAAACACTCTGTATCGGAAACACTCCTGCCT-
AGTTTCAGAATTACACATAAAAATCA AGCCGGCAAACCTATCTTTGCCACTGCC-
ATCTCATTGTTTGCGTCCTGGCCGTCG TTTTACAACGTC SEQ ID NO:320 Late LPZ-311
GCGGACGCCTTACTAAAACGACGGCCAGATGTGTAATGGGGAAAA- TGTGTCATGAT
AGTTGGGTACAAATAACGAGCCACCTGCTCTATGTTTTCGAAG- TTTTCTGTTGGATT
TGTCCGGGTGAGAGAGCGTTCGTTCGTTGCGCGAGAGGGG- CAAAATGCTGAGCG
TGGGGAATTGCCATTGCCGCCCCTGGAAGTGCCGCACGAA- CGCGATCACATTTAA
ATCACCATTTACTTCATCATCACCATGGTTAAATGCAGT- CCCTGCTCCTCAAACAG
GACTTCAGATCCTTCAAGCTCGAAATCTCCGCCTCTG- CTTCCTCGAAGACAAGAC
TCTGTGAGGAGGAAGCGCAGCAGCTGAGCTTAGCGG- ATCTGCTGAAGCCCGGTG
GCCTCGCCCCCGATGGGTTCTCGTACAAGGAGAACT- TTACCATACGCTGCTATGAA
GTCCGAGTTAAACCGCACTGCCACCATTGAGGCG- TCCGC SEQ ID NO:321 Middle
LPZ-312 GACGTTGTAAAACGACGGCCAGC- AACCAAATAAACCCCACATGTGCTCAATGTTTT
AGTATAAAAGGAGATGACTTAAGAGTCATTTCACACACACTTCTATCTTGATTTCTC
CCACTTGTCTTGGGTTTTAGTGGAAGAGAAATCTAGGAGTGGAAGCCCTAGACGTT
GGAGGATAAGAAGGCAACCCTAGAAGGCAGAGCTAACGCTATCCTAAGGCAACCC
TAACGCTATCCTAAGGCGTCCGC SEQ ID NO:322 Late LPZ-314
GCGGACGCCTGCTCAGCACCTGTTATAGTCATTTCTTTTTTCCTTTTTCTCATTTTTG
TCTTTCGAATGACCGCAATGAAATTCAGGCTGCCCAACGCGTTTTGTTTTCACAAT
TAATTTTTGAATCATACGCGAAGATCATGATGAGAATGGTTGTGGAAAAAAACTGTT
TGTAAATATTTAGGTGACCAACAATTTTCATGATTGCAATCTAAAGTTGATAATTGAT
TTATCGGGTCGACATTTGTAATTATTAACACGGAAAATCTGAGGCTTACAATTTTTG
GATTGTAAATATTTAGGTGACGAACAATTTTCATGATTGCAATCTAAAGTTGACAA- TT
GAGTTATCGTGTCGACATTTGTAATTATTAACACACAAAATCTATGAGGCGT- CCGC SEQ ID
NO:323 Late LPZ-315 GCGGACGCCTCATCAATCCATGGTTG-
TACACGCGCCTTCAAAGCGGCTCCTTATG TCGCGCAGCGTCTACTTGTTCCTTG-
AGCGCTTTTCCCTGCTACATCCGCGCGAGCC TCTGTGCAAGGGCCACTGTCTGC-
GCGGTCCCTTTAACTTCGTCGTACTTCTGCTGC
AGCTCACGTGTCTCTATTTCTAAGTGCTATATATTTGGGTCCTCCTGCATAGTAGTG
AACTTCGAACGACTCCTCAAATAGCCAGGTGTAGTCTTTCATTGCACTATTGATCTC
CACTATTCCTGCTATAATGGCGCTAACATGCTGTTCCTTCACCTTTGGCGGAGTG
AAGGCTGCGCCTCTGGAGCTCGGTTATTTGAAGCTGAACCTTGGGCATATCTTC
CTTCACCTCGTGCATCCCCTGCTTCGAGTTTCTGGATGCACGCCTCCACTGGGTCT
TCTGCTGGGATGGGCAACTCTAAGACCAACTGGTATGCGTCGC SEQ ID NO:324 Middle
LPZ-318 GCGGACGCCTTCTTCAATCCATCAGGCCTGATTAATGTATTGACCTTC- TTTGTCTGA
ATGTCATACATTTTTTTCACTGCATCCTTGATCTTCTTCTTGTCT- TGCTTTCTATCCT
TTCTCTTGCTTTCTATCCTTTCTCTGGC SEQ ID NO:325 Late LPZ-320
GACGTTGTAAAACGACGGCCAGCAAAATTGATATAAAGAATAG- ACACATCGACTCA
AATGAAGTGACTCAACAGTTCATAATTCATGTCAGCTTGAA- TGCATGGACATACAC
CCATAAATAGGCAGTTGGGGTCACCCAAAAGAACATAGA- AACATCTCGCATCTCTC
TGAAGAAACTCGGATGGGTACAGGTCTGTGACTTCGC- ATATTTTGAAGGAGCACTC
TCTTGGATAAGTACAATATAGGTACCATCTCGGAC- TCGCCTGAAATCTCGCAAAGA
AGTCTCATTCTCCTCCTTGTTACAGGCGTCCGC SEQ ID NO:326 Late LPZ-321
GACGTTGTAAAACGACGGCCAGAAGCATCA- ATAAACAAAATGACAGATTAACAAGT
TCTCTCTTAATCTTAAGAGAATACATCA- ACATCCAAGTAAAGTCATAACACATTTACA
AAATGGTGCCACGGTATCCATTCT- CTGTAACAAGGTTTTTCTGAAAATAGTTTTCCT
CTTATCTATGTAACTCTTCATAGGGATGCCTGTGTCAACGTGCCATATTCCCAAATT
TGGCCACAATCAAACCTTCCTCATTAGAAGAAACAATCTCTGGTCTAGCTCAAAATT
GGCAAAATTTCCAGCATCTCCCTTTAACATCATTAGAAGGCGTCCGC SEQ ID NO:327 Early
LPS-097 GGGAGATGCTAATTTGAAGCCCTTCTCTGAAGGTGGACAATTCCAGCAG- CAGTGGT
CTAAAGCCCCAATATGGCTATAGAAATTCTTCTGGGGGTTGCACCTA- TGGAAGAGG
GTCGGAGAGGACGAAGCTGTGGATCGCTCTTACCATCTGTGCGGA- AGGTGGTAGC
AGAATTCATTGGAACGTTCTTCCTCATATTTGTAGGATGCGGAT- CTGTCGTTGTTGA
TAAGATAAGCAACGGTTCCATAACTCATCTTGGTGTGTCGC- TTGTATGGGGAATGG
CGGCCATGATTGTAATTTATTCCATAGGCCATATTTCTG- GAGCTCATTTGAATCCTG
CAGTGACGTTGGCCCTTGCGGCTGTGAAGAGATTTC- CATGGGTTCAGGTTCCAGG
CTACATAGTAGCTCAAGTATTTGGATCGATATCTG- CTGGGTTTCTCCTACGTTTCAT
GTTTGGAGAAGTGGCATTCATGGGAGCCACAG- TCCTTCAGGCTCAGAAATGCAGT
CTTTCGCTTTGGAAATTATTACTACGTCATT- GTTGGTGTTTGTGGTTTCTGCAGTCG
CCACTGATACAAAAGCGGTGGGTGAATT- GGGAGGTTCAGCAATTGGAGCGACCAT
CGCAATGAATGTAGGCATATCCGGACC- AATCTCAGGAGCTTCAATGAATCCAGCAA
GGACAATAGGATCCGCAGTGGCTGG- CAACAAATATACAAGCATTTGGGTTTACATG
GTTGGGCCTGTAATCGGTGCGCT- AATGGGTGCAATGAGTTATAACATGATTAGAGA
GACAAAAATGTCCGAAAGGGAGATTATGAAGAGTGGGTCATTTGTTAAGGACATGG
GCTCCAGCGAATCAACAGCATAACAACTTAGAGATTTNTTGCATTCCCGAGACGGT
ATCCAGTGATAGTGGAGAGTAGTCATAATAAGATTTGTGAAAATGTTTGTGTAGATT
AATGTGTAAAATTCAATCCATCAACCATGAAGCGAACTGCATTCCGTTTTTAAATGT
TTATTGGATTTGAATTAATAAACAGCTTATACGTGAAAATCCCTACTTTATGTACGGA SEQ ID
NO:328 Early LPS-098 ACTATAGGGCACGCGTGGTCGACGGCCCGAGCTG-
GTATCCGATGAAGCTAGATTC AATGGTTCAAGTCCTATGAAAGCTAGATTGGAG-
AATTGCAAAGAAATCTAATCTCCG TTAGTTGTCCCAACCACTGACTCGCACCCA-
ATCAGAGTATATTAAAGTTAAAGATTA TATAAAGGTAAATTGAACATTTATAAA-
ATCTTAAATGTATTTTTAGAGTTAAACATTAT
ATAGAATATTTAATGTAGTATAGATATAATAAAATATTAAAAATTAATTTCTCTTTACT
ATCAAGTGAATAAAAATAAAAAATAAATGTAAGACAATATAATAAAAGACTTGTTTTT
AGTGCATTTTTTGGACTCTTCGTTATTGTGTGGTATTGTGTTATTTAAACTGATCTTT
TTACTGTATATATGGATGGGTTACCCATCAAACTTGTGATTTCAATAAATTCCTCCC
GGATTTTAGAGAAATTAGACCATAAAAACTCACGAAAAAAATTTTAGACCATAAAA- AC
TCACGAAAAAAACTTCCCCAAAATCACGCTAAAAACAACTAGATAAAAAAAT- ACCCA
TCTTTGATGATGTGGATAGTGACAGCCTATTCCAAACTATCACCTAAAT-
TGTAAGTT ACATGCATAACACGATGACCTCATCTATACGTTGTGCCAAATAAAG-
GTATGACCGTT CAAACTAAAGAATCAACGAGCTCCAACGCATCTTTTGCTGTGG-
GGGGATTTCTCACG GCTTAACNTTCATGGANCCGATTACCTTNCTANCCAACCA-
AGGGTTTTAACCTGG CAAATNCCAAACCAATTACCAGCTTNACAAATCAACCGA-
GCCGCCCNACCGGGATC ATTTTGGTCAAGTCTCGAAAACNGGCATTGGGTATAT-
GGNATATGGAATTGGAATT GGATCAATGGTAACCTTGGGANAAGCTTAANTTGG-
AAANCCCTTTTTTTTGANGGG GGCCAANTTCCCGNNCCCCCGG SEQ ID NO:329 Early
LPS-099 ATACTCAAGCTATGCATCCAACGCGTTGGGAGCTCTCCCT- ATGGTCGACCTGCAGG
CGGCCGCGAATTCACTAGTGATTAGATGGTAAGAGCGA- TCCACAGCTTCGTCCTCT
CCGACCCTCTTCCATAGGTGCAACCCCCAGAAGAAT- TTCTATAGCCATATTGAGGC
TTTAGACCACTGGTGCTGGAATTGTCCACCTTCA- GAGAAGGGCTTCAAATTAGCAT
CTCCAAGTACATTGATCTATTCTATTCATATA- CATATAACAATGCTGCTCGAGACT
GACAAAATGATCCGTTGGCGCTCGTTGATT- GTTAGCTGTAATTGTTTGGATTGTTCA
GTTAAAGCCTTGTTGGTAGGAGGTAAT- CGGTCATGAATGTTAGCCGTGAGAATCCT
CACAGCAAAAGATGCGTTGGAGCTC- GTTGATTCTTTAGTTTGAACGGTCATACCTTT
ATTTGGCACAACGTATAGATGAGGTCATCGTGTTATGCATGTAACTTACAATTTAGG
TGATAGTTTGGAATAGGCTGTCACTATCCACATCATCAAAGATGGGTATTTTTTATC
TAGTTGTTTTTAGCGTGATTTTGGGGAAGTTTTTTTCGTGAGTTTTTATGGTCTAAAA
TTTTTTTCGTGAGTTTTTATGGTCTAATTTCTCTAAAATCCGGGAGGAATTTATTGAA
ATCACAAGTTTGATGGGTAACCCATCCATATATACAGTAAAAAGATCAGTTTACCAG
CCCGGGCCGTCGACCACGCGTGCCCTATAGTAATCGAATCCCGCGGCCGCCATG
GCGGCCGGGAGCATGCGACGTCGGGCCCAATTCGCCCTATAGTGAGTCGTATTAC
AATTCACTGGCCGCGTTTACACGTCGTGACTGGGAAACCCTGCGTTACCACTTA- AT
CGCTTGAGCACATCCCCTTTTCCAGTGNGTAAAACGAAAAGGCCCCNCCATC- GCCT
TTCAAAAATTGGCAACTGAANGGGAAGGACCCCCT SEQ ID NO:330 Early LPS-100
ATACTCAAGCTATGCATCCAACGCGTTGGGAGCTCTCCCATATG- GTCGACCTGCAG
GCGGCCGCGAATTCACTAGTGATTAGATGGTAAGAGCGATCC- ACAGCTTCGTCCC
CTCCGACCCTCTTCCATAGGTATAAAACCCAGAATTTGGTG- AGCAGGAAGAATTTC
CATAGCCATATTGAGGCTTTACACCACTGCTGCTCGAAT- TGTCCACCTTCAGAGAA
GGGCTTCAAATTAGCATCTCCAAGTTACATGGATCTA- TTCTATTCATATATTTATAAC
AATGCTGCTTCGAGACTGACAAAATTATTTGTT- GGCGCTTGTTCATCGTTAGCTGTA
ATGGTTTGGATTGTTCAGTGTAGGACCAGC- CCGGGCCGTCGACCACGCGTGCCCT
ATAGTAATCGAATTCCCGCGGCCGCCATG- GCGGCCGGGAGCATGCGACGTCGGG
CCCAATTCGCCCTATAGTGAGTCGTATTA- CAATTCACTGGCCGTCGTTTTACAACGT
CGTGACTGGGAAAACCCTGGCGTTAC- CCAACTTAATCGCCTTGCAGCACATCCCCC
TTTCGCCAGCTGGCGTAATAGCGA- AGAGGCCCGCACCGATCGCCCTTCCCAACAG
TTGCGCAGCCTGAATGGCGAATG- GACGCGCCCTGTAGCGGCGCATTAAGCGCGG
CGGGTGTGGTGGTTACGCGCAGC- GTGACCGCTACACTTGCCAGCGCCCTAGCGC
CCGCTCCTTTCGCTTTCTTCCTT- CCTTTCTGGCCACGTTCGCCGGCTTTCCCCGTC
AAGCTCTAAATCGGGGGCTTCCTTTAGGGTTCCGATTTAATGCTTTACGGCACCCT
CGACCCCAAAAAAACTTGATTAGGGGTGATGGGTCACGTAGTGGGCCATCGCCCT
TGATAGACGGTTTTTCGCCCTTTGACGNTGGAAGTCCACGTTTNTTTAATAGNGGG
ACTCTTGGTTCAAAATGGGACAACACTTCAAACCTTTTTTGGGGNTATTTTTTTGA
TTATNAAGGGATTTTTGCCGNNTTTNGGGCCTTTTGG SEQ ID NO:331 Early LPS-101
ACTATAGGGCACGCGTGGTCGACGGCCCCGGCTGGTTTCAATAAATTCCTCC- CGG
ATTTTAGAGAAATTAGACCATAAAAACTCACGAAAAAAATTTTAGACCATA- AAAACTC
ACGAAAAAAACTTCCCCAAAATCACGCTAAAAACAACTAGATAAAAA- AATACCCATC
TTTGATGATGTGGATAGTGACAGCCTATTCCAAACTATCACCTA- AATTGTAAGTTAC
ATGCATAACACGATGACCTCATCTATACGTTGTGCCAAATA- AAGGTATGACCGTTCA
AACTAAAGAATCAACGAGCTCCAACGCATCTTTTGCTG- TGAGGATTCTCACGGCTA
ACATTCATGACCGATTACCTCCTACCAACAAGGCTT- TAACTGAACAATCCAAACAAT
TACAGCTAACAATCAACGAGCGCCAACGGATCA- TTTTGTCAGTCTCGAAGCAGCAT
TGTATATGTATATGAATAGAATAGATCAATG- TAACTTGGAGATGCTAATTGAAGC
CCTTCTCTGAAGGTGGACAATTCCAGCACC- AGTGGTCTAAAGCCTCAATATGGCTA
TAGAAATTCTTCTGGGGGTTGCACCTAT- GGAAGAGGGTCGGAGAGGACGAAGCTG
TGGATGCTCTTACCATCT SEQ ID NO:332 Early LPS-102
ATACTCAAGCTATGCATCCAACGCGTTGGGAGCTCTCC- CATATGGTCGACCTGCAG
GCGGCCGCGAATTCACTAGTGATTAGATGGTAAGAG- CGATCCACAGCTTCGTCCT
TCCGACCCTCTTCCATAGGTGCAACCCCCAGAAGA- ATTTCTATAGCCATATTGAGG
CTTTAGACCACTGGTGCTGGAATGTCCACCTTC- AGAGAAGGGCTTCAAATTAGCA
TCTCCAAGTTACATTGATCTATTCTATTCATA- TACATATAACAATGCTGCTTCGAGAC
TGACAAAATGATCCGTTGGCGCTCGTTG- ATTGTTAGCTGTAATTGTTTGGATTGTTC
AGTTAAGGCCTTGTTGGTAGGAGGT- AATCGGTCATGAATGTTAGCCGTGAGAATCC
TCACAGCAAAAGATGCGTCGGAG- CTCGTTGATTCTTTAGTTTGAACGGTCATACCT
TTATTTGGCACAACGTATAGATGAGGTCATCGTGTTATGCATGTAACTTACAATTTA
GGTGATAGTTTGGAATAGGCTGTCACTATCCACATCATCAAAGATGGGTATTTTTTT
ATCTAGTTGTTTTTAGCGTGATTTTGGGGAAGTTTTTTTCGTGAGTTTTTATGGTCTA
AAATTTTTTCGTGAGTTTTTATGGTCTAATTTCTCTAAAATCCGGGAGGAATTTATT
GAAATCACAAGTTTGATGGGTAACCCATCCATATATACAGTAAAAAGATCAGTTTAA
ATAACACAATACCACACAATAACGAAGAGTCCAAAAAATGCACTATTTACAAGTCTT
TTATTATATTGGCTTACATTTATTTTTTACTTTTATTCACTTGGATAGTAAAAG- AGAAA
TTAATTTTTAATATTTTATTATATCTATACTACATTAAATATTCTATAT- AATGTTAACTC
TAAAAAACATTTAAGATTTATATATGGTCAATTACCCTTATAT- AATCTTTAACTTTAAA
TCCCTGATGGGGGCCAATAANGGTNGGGAAACTAACGG- AAN SEQ ID NO:333 Early
LPS-103 ACTATAGGGCACGCGTGGTCGACGGC- CCGGGCTGGTTTCAATAAATTCCTCCCGG
ATTTTAGAGAAATTAGACCATAAAA- ACTCACGAAAAAAATTTTAGACCATAAAAACTC
ACGAAAAAACTTCCCCAAAATCACGCTAAAAACAACTAGATAAAAAAATACCCATC
TTTGATGATGTGGATAGTGACAGCCTATTCCAAACTATCACCTAAATTGTAAGTTAC
ATGCATAACACGATGACCTCATCTATACGTTGTGCCAAATAAAGGTATGACCGTTCA
AACTAAAGAATCAACGAGCTCCAACGCATCTTTTGCTGTGAGGATTCTCACGGCTA
ACATCATGACCGATTACCTCCTACCAACAAGGCTTTAACTGAACAATCCAAACAAT
TACAGCTAACAATCAACGGGCGCCAACGGATCATTTTGTCAGCCTCGAAGCAGCAT
TGTTATATGTATATGAATAGAATAGATCAATGTAACTTGGAGATGCTAATTTGAAGC
CCTTCTCTGAAGGTGGACAATTCCAGCACCAGTGGTCTAAAGCCTCAATATGGCTA
TAGAAATTCTTCTGGGGGTGCACCTATGGAAGAGGGTCGGAGAGGACGAAGCTG
TGGATCGCTCTTACCATCT SEQ ID NO:334 Early LPS-104
ATACTCAAGCTATGCATCCAACGCGTTGGGAGCTCTCCCTATGGTCGACCTGCAGG
CGGCCGCGAATTCACTAGTGATTAGATGGTAAGAGCGATCCACAGCTTCGTCCTCT
CCGACCCTCTTCCATAGGTGCAACCCCCAGAAGAATTTCTATAGCCATATTGAG- GC
TTTAGACCACTGGTGCTGGAATTGTCCACCTTCAGAGAAGGGCTTCAAATTA- GCAT
CTCCAAGTTACATTGATCTATTCTATTCATATACATATAACAATGCTGCT- TCGAGACT
GACAAAATGATCCGTTGGCGCTCGTTGATTGTTAGCTGTAATTGTT- TGGATTGTTCA
GTTAAGGCCTTGTTGGTAGGAGGTAATCGGTCATGAACTGTTA- GCCGTGAGAATCCT
CACAGCAAAAGATGCGTTGGAGCTCGTTGACTCTTTAGTT- TGAACGGTCATACCTT
TATTTGGCACAACGTATAGATGAGGTCATCGTGTTATG- CATGTAACTTACAGTTTAG
GTGATAGTTTGGAATAGGCTGTCACTATCCACATC- ATCAAAGATGGGTATTTTTTTA
TCTAGTTGTTTTTAGCGTGATTTTGGGGAAGT- TTTTTTCGTGAGTTTTTATGGTCTAA
AATTTTTTTCGTGAGTTTTTATGGTCTA- ATTCTCTAAAATCCGAGAGGAATTTATTG
AAACCAGCCCGGGCCGTCGACCACG- CGTGCCCTATAGTAATCGAATTCCCGCGGC
CGCCATGGCGGCCGGGAGCATGCG- ACGTCGGGCCCAATTCGCCCTATAGTGAGT
CGTATTACAATTCACTGGCCGTCG- TTTTACAACGTCGTGACTGGGAAAACCCTGCG
TACCCACTTAATCGCCTTGGAGCACATCCCCCTTTCGCCAGCTGGCGTAATAGCGA
AGAGGCCCGGACCCGATCGGCCCTTTCCAACAAATTGCGCAACCCTGAATNGGGA
AATGGGCCCCCCCCTNTTACCGGNGCAATTAAACCCCGGGGGGGNGNGGGGGTT
CCCCCCCCCGTGGACCT
[0238]
2TABLE II Clone SE1-SE2 SE3 SE4 SE5 SE6 SE7 SE8 SE9 LPS001 0 249.4
1400.9 827.6 1683.8 2019.4 189.2 4303.9 LPS003 701.2 555.9 2815.2
2445.1 3249.9 3094.7 227.1 3111.6 LPS004 466.1 335.5 2652 2701 2644
2329.6 218.5 2332.4 LPS006 753.1 332.7 3287.3 2964.5 2832.2 2688.9
182.1 1591.9 LPS007 685.2 226 2010.2 1911.3 2600.4 1730.1 181.5
2737.7 LPS008 652.8 274.8 2415 2219.3 2607.1 2294.9 155.7 1292.1
LPS010 558.3 356.1 2667.6 2881.1 2584.3 1573.4 161.7 1041 LPS011
3536.1 424.7 4021.5 3793.8 3590 3182 160.5 1471.7 LPS012 809 408.4
2206.7 2187.1 2282.2 2422.5 462.4 1483.2 LPS013 1211.1 391.6 2294.7
2652.6 2005.4 2167.8 166.8 1570.5 LPS014 2191.9 432.5 2651.8 3013.5
3341.2 3586.7 178.8 3527.1 LPS015 1197.9 306 5651.4 14828.6 20242.8
21558.2 1427.2 34472.3 LPS019 1830.2 334.5 3329 3954.4 4347.5
4658.2 312.1 4743.1 LPS020 675.2 327.8 2258.3 2284.7 2542.7 2321.4
171.9 1609.8 LPS023 451.3 337.5 1401.9 1106.8 1766.2 1842.6 109.6
1365.2 LPS024 4585.8 444.5 3006.3 3431.1 3548.8 3759 157.3 4062.3
LPS025 5102.3 397.1 4322.9 4699.6 5067 4973.2 262.4 5240.4 LPS026
1568.7 285.9 1809.9 1830.4 2829.9 2381.7 164.9 1404.9 LPS027 5499.9
458.4 4853.9 5218.6 2598.4 1756.6 457.9 2375.3 LPS028 4812.9 314.9
2368.8 2616.5 3113.3 3292.4 557 4146 LPS029 4464.6 251.2 2334.4
2058.1 2930.3 3219.3 472 3814.4 LPS030 1142.2 352.5 2519.8 2460.9
2499.8 2634.5 378.3 2147.8 LPS031 1067.7 481.6 3510.8 2799.2 3568.2
3257.2 287.9 2209.7 LPS032 1120.2 332.3 3153.1 3032.4 1769.2 1816.7
146.6 2689.9 LPS036 1498.2 1072.9 4633.6 5524.2 5465.1 6350.7 918
14058.5 LPS037 1890.3 320.9 3719.1 3618.9 4138 4518.1 513.4 5087.5
LPS038 2899.5 310.3 4530 4226.1 4491.6 3969 268.4 4245.3 LPS040
527.4 238.1 1433.4 1611.2 1984.5 1506.5 143.9 1988.7 LPS041 506.1
265.5 1958.9 2843.2 2065.3 2016.2 147.4 2781.7 LPS042 1432.1 1140.3
4379 4973.3 4525.4 4340.8 319.6 3009.6 LPS043 696.9 776.2 3933.1
4894.3 3512.2 3664.7 340.6 3098.4 LPS044 57.8 275.1 3365 4261.2
4773.5 4979.9 974.4 10645.5 LPS045 536.1 211.1 1559.5 1415 1498.5
1584.8 562.1 1912.3 LPS046 796.3 231.7 1023.9 306.4 1417.8 1328.2
83.8 946.4 LPS047 5029.9 518.2 3632.5 4262.1 4755.5 4087.9 386.3
4933.8 LPS050 6333.5 2620.8 5271.4 5242.1 5586.4 5560.1 980.1 11444
LPS051 1378 224.4 2328.8 2221.8 2260.5 2715.1 123.7 3670.4 LPS052
1526.4 267.5 2046 1856.2 2186.5 2416.3 99.3 2010.1 LPS053 4438.3
361.6 4087.6 3959.9 4786.5 3666.8 379.6 4256.7 LPS054 1992.9 269.9
2734.2 2388.1 3143.8 2337.7 177.6 2803.9 LPS055 4587.8 334.4 3488.6
3474 4018.3 3101.6 196.2 4309.4 LPS056 5960.7 1333.7 5338.8 5670.3
5674.4 5533.5 446.4 5593 LPS057 2219.9 301.9 2397.3 2356.1 2218.1
2085.6 184.4 2657.8 LPS058 4070.4 299.9 3485.4 3721.3 4113.8 4142.2
239.8 4945.6 LPS059 8729.3 279.2 3885.7 3636 2720.4 3346.7 165.7
3734 LPS060 4580.2 323.7 3027.8 4713.4 4929.1 5047.5 161 4704.8
LPS061 2831.9 366.8 2392 2327.7 2546.5 1991.8 177.9 3036.7 LPS062
1674.1 353 2711.2 2526.1 1847 1830.3 124.5 3584.2 LPS063 5514.4
419.8 5238.9 5020.3 5417.4 5041 250.1 4812.6 LPS064 7417 3166
5229.5 7497.4 7933.1 10261 1088.3 16829.6 LPS065 5634.9 343.5
5527.8 5099.4 7833.4 5356.6 237.5 4696.7 LPS066 1015.9 244.5 1702.6
1650.5 2895.1 2437.2 128 2514.1 LPS067 2796.8 240.4 3931.5 4810.3
5407.8 5418.3 202.5 9403.8 LPS069 533.4 189.9 1635.8 1816.4 2114.2
1646.8 119.8 3208.8 LPS070 2516.9 240.6 1909.5 2519.6 2156.7 1777.4
186.4 4362.1 LPS071 592.8 196.4 1789.2 2189.2 1981.1 1304.5 127.6
3430 LPS072 444.2 217.6 1422.9 1509 2065.3 2289.9 122.7 2678.8
LPS073 4362.8 273.1 3094.9 3348.1 3771.8 4075.3 137.7 4259.6 LPS074
32072.9 6816.3 33531 25258.9 38176.4 32687.7 14607.1 37529.6 LPS075
7013.9 472.7 4759.7 4933.9 5452.2 5408.7 409.4 5397.1 LPS076 4236.1
362.6 3131.9 2882 3368.5 3354.6 119.5 3141.9 LPS077 2958.7 276.6
4380.4 4862.5 4475.1 4958.7 218.9 4426 LPS078 23685.3 2642.5
35458.6 25869.6 42378.9 33047.1 25402.2 37189.8 LPS079 4794.3 547.8
4628.6 4821.8 5257.2 5277 829.5 5449.7 LPS080 30454 10527 33713.7
23785.4 32590.9 32210.7 16224.4 37659.2 LPS081 30405.9 28677
35358.3 25873 22338.1 31715.3 36436.4 36650.5 LPS083 5040.1 460.8
3251.7 3487.3 2688.9 2565.9 190.5 2979.7 LPS084 2031 298.9 2843.7
2718.4 2352.2 2165.5 164.9 3398 LPS086 3571.7 320.1 2715.8 2648
1989 2528.4 143.9 2969.7 LPS087 3302.3 337.4 4873.1 5695.8 5407.2
5450.6 670.8 18404.9 LPS088 826.8 302.1 2389.2 2871.1 3180.8 2635.2
138.6 3141.5 LPS089 796.4 321.2 1987.7 2640.6 3299.1 2285.1 143.7
3176.6 LPS090 4031 235.9 3867.3 4064.4 4503.3 4798.4 341.7 4697.7
LPS091 2423.3 196.5 2836.8 3101.3 4049.1 4172 295.2 4612.2 LPS092
2914.9 208.5 4005.3 3138.4 3911.6 4036.1 270.4 4842.9 LPS093 793
195.5 1619.2 1331.6 1909.3 1843 147.1 2772 LPS094 1374 221 2205.5
2028.5 2240.9 2632.2 163.3 2849.1 LPS095 728.7 174.1 2022.6 2112.1
2335.8 1264.6 117.5 2957 LPS096 333.3 168.5 1531.9 1393.4 1893.3
869.1 118.3 1691.1 LPZ001 2008.6 185.4 2535.9 2937.9 3472 1981.8
118.9 2421.7 LPZ002 3529.3 384.6 4579.3 4474.6 3236.7 3855.8 313.8
3237.5 LPZ003 4076.8 275.4 2651.2 2966.7 2829.2 4177.4 378.5 4369.7
LPZ004 5595 687.4 5468.2 5615.9 5243.6 5699.6 601.6 5889.9 LPZ005
5680.5 3353 34994.7 26121.9 42555.1 33144.5 16193.7 37798.2 LPZ006
1199.8 299.4 3013.7 3099.8 3517.3 3397.1 140.6 3370.8 LPZ007 1159.1
462.2 3292.7 2992.5 3121.4 2936.7 235.5 3238.6 LPZ008 1874.3 237.7
3110.8 3236.7 2516.5 3182.2 325.3 4330.1 LPZ009 3331.1 296.3 2348.5
3414 2478.2 3309.5 348 5658.1 LPZ010 3216.3 1186.8 4977.3 5024.7
4564.4 4992.4 442.6 4454.5 LPZ011 4613.4 910.9 4510.7 4515.7 3729
4357.3 371.4 4695.9 LPZ012 1531.5 469.5 2915.3 2611.1 2012.3 3481.4
270.3 3804.3 LPZ013 3495.1 268.8 2125.9 2584.7 3194.7 3787.4 125.1
4929.6 LPZ015 2040 257.6 1971.1 2966.7 2191.1 3056.7 227.1 4156.6
LPZ016 5307 2761.1 8451.7 17219.7 22792.7 15567.3 1073.6 35074.1
LPZ017 2476.4 354.3 3175.5 4330.8 4496.2 4061 273.2 5328.9 LPZ018
3929.4 417.5 12420.2 14916.1 18116 17637.5 2541.6 31981 LPZ019
5404.2 427.3 32190.3 24710.4 42102.7 32342.6 19528 36969.5 LPZ020
576.9 142.9 1451.4 1505.4 3534.8 2679.8 210.9 3046.2 LPZ022 1408.2
155.2 2406.7 2845.7 3042.5 3074.8 189.9 3829.2 LPZ023 562.1 152.8
2096.7 1710 2045.5 2078.9 200.8 2874.3 LP2024 496.7 158.1 1681.3
1264.7 2102.9 1857.1 132.1 1818.4 LPZ025 5431.3 464.1 13492.2
9726.2 11911.5 13462.8 1262.5 11780.6 LPZ026 1663.2 139.7 2464.8
2760.1 3113 2219.4 159.1 3183.5 LPZ028 5029 190.7 5367.2 5339.8
5483.9 5205.5 482.3 5565.9 LPZ029 961.3 119.2 1805.4 1989.6 2298.5
1998.4 126 2576.9 LPZ030 1457.4 177 2444.7 2687.5 1966.4 1857.2
178.5 3312.8 LPZ031 3092.8 361.7 3564 3925.3 4627.8 5171.4 506.7
5920.5 LPZ032 1906.5 156.8 5542.3 24342 42917.8 33386.1 30058
37998.6 LPZ033 12934.5 354.7 5280.1 7301.2 5638.9 9238.7 375.4
15843.5 LPZ034 1307.4 177.5 1737 2208.4 3213.1 1984.1 150.2 3228.3
LPZ035 556.5 201.9 880.2 1280.1 1654.5 915.1 74.1 1422.1 LPZ037
1356.8 269.7 2072 3110.5 2912.8 2488.2 211 4119.3 LPZ038 4027.9
426.9 5639.9 5872.3 5476.8 5614.6 796.8 5583.3 LPZ039 5059.1 550.6
3807.9 4393.8 3825.6 3889.8 342.2 5164.2 LPZ040 1226.1 236.5 1566.4
1889 1679.1 2263.6 140.6 3331.1 LPZ041 944.2 219.3 1629 543.1
1148.2 1416 90.2 2524.6 LPZ042 570.6 206.1 1129.5 806.5 1448.8
1423.1 75.1 2013.8 LPZ043 1190.2 236.7 1878.8 1024.4 2834.6 2767.4
241.7 3236.2 LPZ045 5315.3 465.7 4933.2 5580.2 5151.1 5205.1 557.3
10754.3 LPZ047 859.5 285.2 1606.2 2099.3 2059.4 1992.6 68.3 3054.8
LPZ049 3232.7 108 1278.6 2834.2 3657.8 3944 244.2 5459.6 LPZ051
3048.1 146.9 2373.2 2067.3 2745 2383.2 179.1 2837.6 LPZ053 2580.3
135.6 2625.8 2088.7 2468.5 2297.2 156.8 3001.4 LPZ054 1838.1 159.5
2657.8 2759.7 2658.1 2224.7 170.4 3444.2 LPZ055 2181.8 151.1 2381.2
2262.7 3228.3 2983.9 139.3 2673.9 LPZ056 4028.3 219.5 2884.6 3416.6
3779.6 3789.9 208 4518 LPZ057 1470 121 1676.5 1629.6 1702.7 1703
112.2 2272.1 LPZ058 1923.3 122.5 2453.5 2169 3127.3 2465.4 160.6
3319.6 LPZ059 1760.4 113.8 2180.6 1832.4 1997.2 1530.8 174.4 3366.6
LPZ060 3296.4 139.3 2571.1 2250.2 2721 2976.9 221.3 3896.5 LPZ061
2495.6 182.8 2663.9 2235 3265.9 4227.1 498.1 4915.1 LPZ062 1992.7
194.9 3296.7 3975.8 3861.5 5642.6 497.6 5606.2 LPZ063 2167.1 145.9
2733 1843.9 3066.6 4961 305.6 4773.2 LPZ065 5641.2 251.7 13690.3
9269.2 8562.8 13254 986.3 9554 LPZ066 6307.3 652.4 12630.8 6968.4
4918.9 5062.2 400.7 5456.8 LPZ067 10838 1548.1 16986 11776.8 5633.2
7054 1014 15262.2 LPZ069 1481.9 209.6 2239.8 1480.9 2496.7 2542.4
250.5 3717.2 LPZ070 1932.5 263.8 1895.1 2221 1555.9 1570.4 145.5
3471.3 LPZ071 3672.6 378.6 4185.5 3050.5 4166.8 4246.2 553.7 5333.4
LPZ072 744.5 210 1210 676.7 1420.2 1393.4 95.8 1997.1 LPZ073 1997.9
235.9 2275.1 2141.7 2613.2 1989.9 170 3489.4 LPZ074 1375.9 237.4
1899.1 1787.3 2472.9 1623.7 125.6 2435 LPZ075 831.4 247.9 1536.4
1773.1 1886.9 920 80.6 1053.5 LPZ076 345.7 251.8 854.8 564.6 1747.1
526.2 55.9 1058.3 LPZ077 2466.3 102.2 949.4 820.9 3093.9 3179.6
202.9 3314.8 LPZ078 3102.1 197.1 3654.2 3261 4204.3 4433.6 400.8
5559 LPZ079 1584.4 108.3 2389.2 2243.3 2624.8 2677.1 208.3 3675.6
LPZ080 12206.5 2043.1 25021.4 8579.5 11707.8 8717.6 1172 18663.9
LPZ081 1368.7 103.6 1902.8 1349.9 2166.1 1597.7 103.5 2709.6 LPZ082
2601.3 140.3 3264.3 2853.9 2799.6 1742.3 251.1 4288.2 LPZ083 1311.9
76.7 1622.4 1071.1 1733.9 1878 104 2007.7 LPZ084 9974.7 801.3
14255.3 8399.1 5763.9 8852.9 542.2 5714.3 LPZ085 4609.8 158.4
3923.3 3729.7 4082.8 3867.3 219.3 4075.1 LPZ086 10874.1 987.4
19189.5 8284.6 5646 9109.8 1116.4 14988 LPZ089 3505.8 211.6 4010
3430.6 3762.1 3770.8 224.3 5341.2 LPZ090 5780.9 581.8 13217.4
6303.4 4694.8 4779.9 425.2 5408.9 LPZ091 5316.1 148.4 2263.4 2139.8
2382.2 4067.2 256.8 14732.6 LPZ092 5448.7 209.4 3631.6 4152.7
2934.1 3403.7 174.9 4943.6 LPZ093 1169 159.4 2097.9 1187.4 2050.8
2350.7 109.4 2605 LPZ094 1245.5 139.7 1547.5 1650.5 1875.2 2009.9
80.2 2376.9 LPZ095 711.2 177.9 900.9 1253.3 1013.8 1395.3 48 1586.1
LPZ096 2122.2 249.7 2929.3 3271.3 2132.9 2224 232.8 4443.8 LPZ099
4306.4 211.2 2603.1 2144.4 3479.2 3488.5 138.1 4085 LPZ100 3373.5
297 3941.3 3149.6 3790.4 3857.5 443.8 5028.1 LPZ101 3007.7 272.4
3546.9 2291.3 4299 3232.1 306.1 4819.6 LPZ102 2092.7 324.7 3167.5
2109.3 3524.3 2829.4 279 4297.4 LPZ103 3602.1 285.7 2923.3 3112.9
2812.9 1318.3 87.9 1739 LPZ106 1359.7 305.1 2680.3 2391.6 2838.5
2097 173.7 3009.6 LPZ107 28560.8 4989.5 20821.7 17880.4 39173.1
27035.1 11973.3 36123.4 LPZ108 4136.8 179.4 4259.8 4978.2 5553.2
4862 837.2 5597.5 LPZ109 3708.3 202.4 3842 3510.4 4638.4 4453.7
469.5 5107.4 LPZ110 4557.2 291.4 5020.6 4801 4487.4 4481.1 552.3
5484.2 LPZ111 1625.6 130.9 2242.1 1982.7 2740.6 2455.4 164.6 3722.3
LPZ112 2887.4 195.8 3813.2 3759.4 3984.8 4167.1 409.7 5461.8 LPZ114
5029.5 213.4 5016.7 4678.8 5036.9 5168.1 302.1 4316 LPZ115 24434.4
2637.1 27958 23684.2 41104.3 30920.9 2153.9 36902.6 LPZ116 8682.9
235.7 5647.3 5316.6 5805.6 9313.7 466.6 16018.9 LPZ117 30879 4843.7
36277.1 24358 24673.1 20545.7 4669.9 5652.6 LPZ118 4023.6 171.1
3743.5 4568.2 3845.4 3783.9 254.3 4782.5 LPZ119 2580.4 114.1 2507.2
3114.1 2544.6 1963.8 127.6 3195.4 LPZ120 1998.8 157 1987.2 1503.1
2331.8 1805.1 131.5 3522.3 LPZ122 2041.4 119.6 2145.6 2430.9 1998.6
2171.8 101.3 2677 LPZ124 2795.6 185.4 2980.4 2672.5 2495.2 3459.4
173.1 3081.5 LPZ126 2559.7 181.8 2560.1 2349.8 3500.6 2362.1 224.9
3646.9 LPZ127 1993.5 169.1 3161 3180.8 3382.5 3321.3 180.6 4058.4
LPZ128 2866.7 263.2 3556.8 3597.4 3545.7 3813.8 306.7 4071.3 LPZ131
1993.5 171.7 1983.9 2069.6 2565 2607.2 80.3 2527.8 LPZ133 2446.7
290.4 3218.6 2847.2 3830.1 2889.5 245 4252.4 LPZ136 1952.3 281.1
2956.9 1870.6 3167.6 2680.6 215.9 4291.6 LPZ137 2833.8 281.9 3264.4
2350.2 3874.4 3532.8 420.8 4935.3 LPZ138 2932.9 1791 5211.5 4502.1
5409.9 4832.8 543.1 4741.3 LPZ140 2284.7 337.4 3680.2 2810.9 3196.1
3191.2 271 4613.6 LPZ141 4726.2 368.5 4792.5 4412.5 5368.1 5466.3
722.1 4956.4 LPZ143 25290.6 2692.2 35967.9 25679.9 43668.3 32612.1
25456.9 36344.4 LPZ144 2620.9 286.6 3948.7 3394.6 4505.7 4142.8
488.7 4776.7 LPZ145 3472.5 171 3949 3194.2 3430.5 3539.9 327.9
4487.2 LPZ146 2612.8 127.3 2482.4 2080 3000.8 2979.1 135.1 3391.3
LPZ147 2447 106.3 2855.1 2237.7 3134.2 2841.8 261.6 4388.1 LPZ148
2036.8 77.7 2559 1932.3 4296.1 4699 359.6 3982.3 LPZ149 5720.7
267.4 5377.3 5408.2 10999.7 5717.7 1078.9 13033.2 LPZ150 5861.7 772
35541.7 26314.8 44633 33238 13126 37853.6 LPZ151 5550.3 3499.3
9012.8 8380.4 11968.1 5716.5 715.8 5536.9 LPZ152 4746.6 352.8
5169.3 5647.7 5384 5394.2 408.5 5382.1 LPZ153 21881.2 2773.2
14738.2 15979.5 16996.8 15756.8 2388.5 30812.9 LPZ154 4869.8 265.9
3244.3 3497 3948.6 3703.3 303.8 4119.1 LPZ155 3904.2 1596.3 5078.5
5482 4631.7 5314.1 553.4 4112.9 LPZ157 4726.5 1732.8 5427.1 5369.5
5213.3 5705.9 756.4 5462.2 LPZ158 15297.4 3817.1 17993.9 17405.3
25168.8 22056.6 2337.4 22375.3 LPZ162 5725.8 4204.8 10380.1 11364
17948.2 14250.8 1934.6 10535.5 LPZ165 5615.2 666.7 5274.6 5486.6
5560.3 5310.9 637.1 5405.9 LPZ166 5889.1 2603 9503.5 10943.7
13743.3 14080.4 1772.5 5772.8 LPZ167 5347.2 1948 5708.9 6769.6
5742.3 5347.9 370 5279.7 LPZ169 3043.8 267.4 1976.6 2851 3451
2451.2 189.8 3420 LPZ170 3507.3 301.5 3532.3 3391.4 4481.4 3398.7
130.2 5604.2 LPZ171 3762.3 780.7 4554.8 4311.7 4936.4 4511.3 398.6
5030.5 LPZ172 5098.2 947.4 5550 6287.9 5135.4 5323.9 1242.5 8539
LPZ173 22580.5 3313.9 35486.9 24974.9 42874 31828.8 26531.8 36066.9
LPZ174 4115.7 221 5241.5 4262.4 5765.8 5554.9 872.5 4815.2 LPZ175
4388.3 1360.6 5563.7 5504.9 5165.3 5182.9 583.2 4602.3 LPZ177
1371.6 94.5 2119.4 2218.6 2730.7 2431.7 143.2 2893.2 LPZ179 3643
195.1 4409.9 4898 5458.3 5319.8 797.9 5677.7 LPZ181 5573.3 215.9
4799.6 5272.2 5825.3 5554.4 1573.5 13689.7 LPZ182 4118.9 107.6
3491.5 3182.1 4617.5 4543.6 478.1 5527.8 LPZ186 5792.2 325.5 4965.1
5182.6 12373.6 11191.4 1804.9 37336.4 LPZ189 33820.3 5188.5 30941.4
24955 43453.2 33115.2 17929.3 38055.6 LPZ194 2807 151.1 2915.9
2955.1 3306.8 3120.2 142.9 4101.6 LPZ195 5345.7 532.7 5597 5628.7
5540.4 5491 545.7 5756.7 LPZ196 4805.1 3512.9 5183.2 6968.6 5465.4
5052.4 786.4 5694.5 LPZ197 7268.6 153.8 5398.9 5673.5 13582.6
15111.9 3499.6 34684.8 LPZ198 7208.3 210.6 5800.8 8043 5439.2
5183.6 409.3 5042.7 LPZ199 3058.3 186.1 2749.6 2667 3713.6 3704.3
243.4 3917 LPZ201 7175.3 236.7 4827.6 5029.9 5523.4 5802.2 1981.9
14614.5 LPZ202 3603.3 1113.9 35531.5 26035.1 44762.8 33837.3
63521.8 38225.4 LPZ203 4325.4 424.4 5517 5387.3 9934.8 5662.1
2104.8 9370.7 LPZ204 32355.9 34690 36443.6 26004.3 44546.1 33680.5
55702.6 37890.3 LPZ205 4904.1 519.6 5162.4 5398.7 5427.6 5325.6
281.4 5770.2 LPZ206 3504.4 319.8 3124.8 4561.7 4192.2 3899.9 255.7
5489.9 LPZ207 32035 24978.7 34825 23371.8 42639.9 32686.4 30672.2
37674.8 LPZ208 25174.6 3118.6 14244.4 13906.3 16694.7 21111.9
2190.1 34542.4 LPZ210 3885.1 422.3 3895.8 4551.5 4205.7 5108.7
258.6 5514.3 LPZ211 2569 176.7 3689.2 2943.5 4001.9 3860.9 250.2
3113.1 LPZ212 5988.8 1244.3 32684.4 11154.1 19853.4 13654 618.3
10736 LPZ213 3406.9 106.8 3964.1 3876.6 4236.4 4294.4 274.2 4874.6
LPZ214 1668.3 55.3 2136.3 2394.8 2390 2269.3 105.8 3436.2 LPZ215
5019.8 139.3 5020.1 5024.8 11013.9 13747.1 1991.7 36930.6 LPZ216
3336.8 1085.4 35895.6 26245.3 44980.1 33834.4 64482.7 38238.1
LPZ217 23512.1 26363.3 36065.8 24685.4 43193.4 31422.4 21462.1
35990.6 LPZ219 4011 256.9 3193.5 3326.3 4509.5 5258.5 455.2 5841.9
LPZ220 8696.5 2383.3 5064.7 5171.3 4923.7 5340 951.7 17530.6 LPZ221
1221.4 83.1 1201.8 707.6 1556.5 2083.9 182.1 3948.8 LPZ222 1885
146.1 2834.7 2253.2 2557.7 3382 196.7 4225.1 LP2223 1048.5 121.2
2339.6 2642.1 2663.8 3573.5 383.5 4579.2 LPZ224 3190.6 118.8 3049.8
2833.2 4373.8 5139.9 858.6 5285.7 LPZ225 25428.2 4079.7 35724.5
25423.6 43300.5 32574.8 38888.3 37219.9 LPZ226 1044.9 130.4 1776.9
1210.8 2757.7 3388.5 326.6 3520.3 LPZ227 1078.3 133 1461.7 973.5
7032.1 9452.8 2043.5 4705.3 LPZ228 3961.6 213.5 3373 4050.7 5575.6
10714.4 2428.6 5928.9 LPZ231 3475.2 230 4096.3 3841.1 5009.3 5690.3
959.9 5514.3 LPZ233 2404 218.2 2170.9 1531.4 4362.7 4198.1 673.7
3350.1 LPZ234 1688.3 312.3 1887.6 1486.5 4228.6 4715.6 724.8 3170.1
LPZ235 2661.6 199.9 2422.2 1852.6 3078.9 2886.6 98.2 3143.5 LPZ237
3174.5 324.2 3032.5 2988.2 3931.1 4587.9 314.4 4588.3 LPZ239 4061.3
309.3 3175.1 2932.1 4131.7 3892.6 122.3 5083.5 LPZ240 3799 316
3730.6 3314.6 3379.8 3538.8 212.4 4784.5 LPZ241 2559.2 62.1 2610.4
1794.5 4165.6
3754.4 134.8 2915.1 LPZ242 29360.5 3262.9 35254.6 25196.9 43028.8
31468.2 7308.4 36768.4 LPZ243 3405.3 88.8 3015.7 2683.4 3678.7
2990.6 121 4001.4 LPZ244 4856.9 483.6 4842.2 5235.3 5317.6 5432.1
205.4 5712.9 LPZ246 1274.8 65.9 2301.7 1922.8 4332.2 4628.8 672.1
4232.4 LPZ247 3894 69.8 2522.8 3389.9 4451.4 4937.1 939.3 5522.8
LPZ248 3016.7 268.6 2883.2 3805.2 3791.7 3777.6 487.1 4585.6 LPZ249
5224.1 138.3 3524.5 4091.2 3022.4 3393.2 149.9 4101 LPZ250 1060.6
46.5 1400.9 1246.9 1419.5 1411.2 118.8 2908.4 LPZ251 1336.8 248.5
1354.6 1049.3 657.6 924.3 70.3 2064 LPZ255 3787.8 171.8 4801.8
5076.3 4608.1 4965 340 5636.6 LPZ256 536.6 61.6 865.5 971.6 1130.8
1327.7 82.7 936.9 LPZ257 844.5 112.6 1507.4 1537.8 2337.9 2745.8
341.5 1610.7 LPZ258 2588.5 142.3 3443.1 2902.2 4576 4976.4 1182.9
3619.6 LPZ260 837.7 113.7 1677.9 944.3 1217.6 1286.6 170 2928.2
LPZ261 981.3 132.1 1499.6 743.4 1590.8 1953 67.2 1652.1 LPZ264
4559.3 231.3 4348.5 2856.3 4869.2 5179.7 412.3 4698.3 LPZ265 21063
2793.9 26928.5 12365.5 13816.5 12134.4 691.6 17954.8 LPZ266 1642.4
130 1767.5 1463 1633.8 1410.5 59.4 1444.5 LPZ268 2451 114.2 2803.3
2495.4 3126.5 3433.7 79 4261.3 LPZ269 15670.7 3660.5 35782.4 21720
40375.2 31597.3 2024 35213.6 LPZ270 3541.2 240.8 3803 3132.4 4827.8
5213.6 79.9 5473 LPZ271 5590.7 677.2 5465.4 5197.1 5703.4 5615.2
309.2 5732.7 LPZ272 27369.6 3445.9 35824.6 22832.6 40684.8 27398.4
1732.9 37016 LPZ273 1107.3 46.1 456.7 336.3 1879.3 1654.1 65.4
971.9 LPZ274 3936.2 114.5 3192.3 3024.1 4983.3 4907 293.9 4933.5
LPZ275 2567.2 42.9 1760.4 2091.8 3656.5 3800.5 77 2585.4 LPZ276
560.9 32.9 1075.4 1878.9 1889.9 1766.2 66.8 1294 LPZ277 423.7 34.6
1199.1 1169.8 1376.8 1383.9 91.9 1123.7 LPZ278 323 39.7 937 382.9
770.1 935.2 66 1403.4 LPZ279 965.9 70.7 1907.5 1368.2 1783.7 1603.9
133.2 2438.8 LPZ280 390.7 19.6 175.4 42.3 464.9 631.5 29.9 2074.9
LPZ281 84.3 8.2 0 0 0 0 9.7 0 LPZ282 1849.7 28.4 315.1 34.2 664.3
1097.3 21.5 1229.5 LPZ283 10678.6 329.2 5134.7 5311.1 4772.3 8591.1
226 9633.6 LPZ284 996.1 39.8 236 147.2 2349.5 981.1 26 719.7 LPZ286
563.8 77.1 1031.1 945.9 1347.4 1601 81.2 1303.6 LPZ287 1045.7 123
2057 1475 1730.9 3003.6 149.9 2493.5 LPZ288 1201.7 116.2 1797
1448.8 1648.3 670.4 80.6 3700.4 LPZ289 1922.3 113.3 2515.4 3395.3
3460.7 3369.4 70.8 2183.8 LP2290 14629.5 3945.8 34659 24047.3
40474.8 27786.2 1348.2 27566.4 LPZ293 4364.8 385.4 4664.2 3170.9
4321.6 4789.8 74.6 5095.8 LPZ294 564.7 171.4 1257.5 705.2 1357.7
1610.2 18.6 2027.6 LPZ295 823.1 97.3 2102.7 1056.2 2899.7 2698.3
39.4 2448.2 LPZ297 5273.4 169.1 5229 5074.4 5727.8 11512.9 423.2
10966.8 LPZ299 1564 161.1 1743.9 1752.3 2764.1 2660.5 63.5 2791.9
LPZ300 3068.3 205.4 2406.8 1881.8 2898.6 2758.4 0.2 2007.2 LPZ301
1979.7 233.1 3207.1 2109.3 4343.5 3713.8 40.4 2690.6 LPZ303 509
32.7 281.3 877.7 893.1 751.5 30.1 1373.8 LPZ304 2531.1 289.3 3809.4
3406.7 3674.8 3517.4 158 2652.7 LPZ306 22632.7 2861.2 34933.8
25435.6 40453.9 30906.9 1505.9 34032.8 LPZ307 2604.4 1395.2 4780.6
6945.3 4419.2 4416.9 232.6 4299.8 LPZ308 1093.9 60.5 2028.1 1751.6
1770.8 1891.9 92.4 3245.8 LPZ309 286.1 26 480.4 378.4 589.6 731.4
38.4 1062.4 LPZ310 2284.1 129.5 1622.7 1091.7 1207.1 3089.4 101.2
3624.9 LPZ311 3309.9 43.6 2782.6 2956.3 2828.8 4446.7 95.8 5593.4
LPZ312 446.3 52.7 1577.7 1221.4 542.2 518 56.1 1952.6 LPZ314 378.6
26.9 333.9 355.8 682.2 701.2 61.7 732 LPZ315 3897.5 115.2 2611.9
3145.8 4296 5240.2 151.3 4499.3 LPZ318 9709.6 767.1 19964.9 15678.3
20611.8 19600.2 475 18079.9 LPZ320 1126.7 82.8 1215 1002.7 1502.5
1555.3 67.5 2964.4 LPZ321 2944.7 85.4 2590.7 2597.6 2550.3 2962.7
72.1 5481.4 Clone ZE1 ZE2 ZE3 ZE4 ZE5 ZE6 ZE7 ZE8 ZE9.1 LPS001
369.9 369.9 369.9 369.9 369.9 369.9 369.9 369.9 369.9 LPS003 600.3
363.9 0 243.7 1565.3 2624.5 1942.7 242 1892.5 LPS004 522.3 254 0
74.6 907 2638.8 1933.6 274.9 4209.2 LPS006 444.6 161.2 0 174.6
793.6 2651.4 1991.5 206.5 598.8 LPS007 528.9 136.3 0 244.9 1623.3
1202.1 2044.7 245.1 213.9 LPS008 534.5 215 0 281 1231.2 783.4
1760.3 178.5 832.4 LPS010 469 183.6 1.3 240.1 947.7 591.6 2208.3
161.2 482.6 LPS011 468.7 93.3 0 142.3 1544 1021.5 2334 254.6 1223.7
LPS012 511 278 17.7 197.2 2129.6 68.9 1362.7 478.9 960.4 LPS013
478.2 407.1 192.1 235.6 2470.6 9 1163.9 885.7 1109.4 LPS014 579.7
369.1 0 272.7 2799.6 1525.7 2222.4 606.9 2638.6 LPS015 419.7 254 0
2380.7 7188.1 4998.4 16519.6 5245.1 15550.4 LPS019 1068.4 279.6 0
396.2 3848.5 3074 3866.9 959.1 3664.7 LPS020 314.2 109.1 0 102.7
2036 234.4 1504.2 319.6 1053.2 LPS023 364.9 104.2 0 100.7 1151.8 0
1253 175.5 570.5 LPS024 804.7 213.8 0 346.3 3248.5 2523 2722.6 915
1987.4 LPS025 1374.7 407.8 0 857.1 4731.2 2584.2 4119 1138.4 2458.4
LPS026 337.6 86.1 0 100.2 1242 0 1052.9 242.9 988.9 LPS027 440.5
182.5 0 118.5 1318 691.2 1274.1 226.1 385 LPS028 369.5 166.2 0
168.7 2587.7 2597.8 4035.5 565.9 1883.1 LPS029 323.4 141.9 0 165.3
2524.3 2147.2 3031.3 567 2263.9 LPS030 362.3 226.5 0 169.6 1528.2
422.9 1236.7 239.2 1049.1 LPS031 591 536.7 4.9 383.6 1768.3 850.4
1013.3 399.8 781.4 LPS032 443.9 327.3 0 328.1 3200.9 1880.1 1832.8
265.6 1391.6 LPS036 1093 781.8 24.5 680.8 3911.3 3750.9 3746.7
661.8 3856.4 LPS037 501.6 180.4 0 200.9 2664 2369.8 1960.5 339.9
2892.9 LPS038 1180.1 471 155.9 1679.7 4392.3 2103.5 3019.9 800.3
2819 LPS040 398.8 108.6 0 103.9 1030.7 195.9 1566.1 144.4 682.1
LPS041 384 153.8 0 149.7 989.2 1257.5 2235.4 143.3 1228.4 LPS042
1381.9 951.7 44.9 716.1 3682.1 2755.7 4011.7 508.5 2963.4 LPS043
1211.6 704.8 74 613.6 3494.4 2435.3 3362.1 391.4 1544.7 LPS044
361.3 100.2 0 142.1 2244.4 3031.8 2653.5 393.4 1620.1 LPS045 285.7
75.2 0 64.4 856.5 223.5 1616.1 216.8 609.7 LPS046 325.8 217.2 0
70.2 1758.6 0 1280.9 284.9 1115.5 LPS047 2041.3 1347.8 768.4 1080.5
4169.9 3927.9 4263.5 1831.6 4804.9 LPS050 3226.4 3356.4 6064.5
3347.5 9841.4 3046 5362.2 2924.8 5821.2 LPS051 377.1 96.4 0 156.8
2452.6 2286.5 3035 396 2238.1 LPS052 330.1 80.1 0 162.6 2418.1 0
2097 352.5 1677.3 LPS053 402.6 160.1 0 146.3 2249.5 56.9 1986.1
464.3 1349.6 LPS054 497.4 147.5 0 184 2188.4 379.1 1976.1 308.6
1558.9 LPS055 1168.2 645.7 0 354.9 3901.1 1476.5 2607.5 774.8
3026.1 LPS056 1549.5 1243.3 37.9 752.6 4770.9 3403.7 4086.8 1204.2
4958.4 LPS057 387.5 154.3 0 262.7 2612.2 502.6 2317.5 365.1 1418.8
LPS058 671.2 198.9 0 434.8 4189.9 2258.6 3366.3 586.3 2190.5 LPS059
726.2 207.5 0 304.6 2974.8 2054.3 2712.8 395.1 1331.6 LPS060 534.2
215.8 0 221.7 2896.9 718.3 2693.3 477 2474.9 LPS061 530.8 369.4 0
204.3 1801.1 1286.4 1533.6 298.7 1327.2 LPS062 407.4 305.2 0 226.4
1509 0 1413.1 212.5 954.6 LPS063 619.4 280.8 0 282 3987.4 1805
2589.9 642.1 1650.4 LPS064 3689.2 4982.4 10201 3080.3 8359.8 3622.3
8304.6 2997 13781.1 LPS065 466.4 189.7 117.3 817.1 4336.3 2332.6
4393.4 1092 3866.8 LPS066 269.5 104.6 0 131.4 1006.2 76.7 1834.6
185.9 668.5 LPS067 426.4 179.7 49.3 341.3 4153 4077.7 5101.3 1195.9
3894.1 LPS069 367.8 136.7 0 128 1456.6 0 2685.6 308.7 1234.3 LPS070
438.5 137.3 0.4 111.6 1932.5 25 3005.3 210.4 721.5 LPS071 283.9
83.2 8.5 109.2 1831.5 0 3634.2 302.8 708.3 LPS072 301 147.2 5.7
132.9 1600.8 592.3 3051.5 331.5 1173.5 LPS073 692.1 485 251.5 497.9
4205.3 2827.4 3777.9 740.1 3882 LPS074 36280.3 66359.2 63362.2
44047.1 47176.9 20938.9 64534.5 33666.4 78457.8 LPS075 3204.8
1250.6 650.9 1033.7 4976.3 4377.1 4632.6 1617.1 5570.9 LPS076 434.7
127.9 0 204.7 1731.3 419.1 2737.8 298.8 2175.1 LPS077 416.6 107.6 0
327.2 3360.7 1950.5 4020.1 609.1 3713.9 LPS078 5164.5 1194.3 906.3
6556.7 20779.8 7364.3 28847 8680 22339.1 LPS079 1317 501.9 304.2
893.4 5047.6 3196.6 4887.5 1058.8 4992.1 LPS080 27721.4 56038.2
70896.6 26826.4 43426.4 20557.3 47819.2 16935.5 68145.5 LPS081
36397.3 66337.3 48195.9 41685.3 46187.5 20628 66138.1 31620 78253.7
LPS083 844.6 534.2 123.6 305.3 3724.2 1699.7 2524.6 583.8 3266.4
LPS084 665 249.4 0 334.5 2570.6 1491.7 2893.2 342.8 2061.7 LPS086
456.6 155.8 0 165.1 1962.7 754.5 1931 130.2 2176.5 LPS087 967.5
450.7 17.2 633 4238 3720.3 5373.9 1754.7 19094.1 LPS088 468.4 276.1
0 151.1 1109.4 0 1779.7 302.8 2497.2 LPS089 329.2 316.9 0 133.6
988.7 0 1619.6 320.7 1616.9 LPS090 478.9 272.3 0 218 4486.4 2182.3
2923.1 584.6 2640.7 LPS091 385.7 177.9 0 290.7 2923.3 2008.6 2453.2
441.5 2246.6 LPS092 396.3 164 0 345.2 2249.1 1219.1 2906.7 413.9
1589.3 LPS093 308.3 164.5 24 98.7 262.3 427.8 2140 175.4 661.4
LPS094 331.6 179 54.2 146.4 773.3 948.2 1729.1 116 1030.7 LPS095
363.7 157.7 46.3 142.5 967.7 341.4 2639.7 199.3 1055.4 LPS096 266.9
90.6 0 59.3 676.5 0 2616.2 136.5 215.6 LPZ001 270.9 49.7 21.1 121.2
1958.2 496.8 4495.7 325.6 557.7 LPZ002 491.7 231.8 157.9 345.8
1929.4 1183.4 3243.9 305.5 932.6 LPZ003 632.6 407.2 342.3 343.5
2630.6 2108.3 3212.8 423 4663 LPZ004 2034.4 2260.7 1487.4 1442.5
5730.7 2135.4 5424.1 1804.4 5786.1 LPZ005 6301.3 4683.6 2801
10127.5 31972.9 7747.5 51335.5 17767 52067.8 LPZ006 471.7 131.2 0
179.4 1964.1 1552.2 2977.9 333.5 2954.9 LPZ007 584.6 383.7 69.3
294.3 1424.1 531 2664.9 265 3021.7 LPZ008 325.4 87.5 14.4 176.8
1966.6 1588.5 2420.1 199.5 3481.2 LPZ009 451.3 281.6 143.4 362.3
3149.2 4280.4 3006.5 357.5 4395.9 LPZ010 1324.5 1442.8 621.8 931.8
4310.4 3238.7 3926.9 617.4 4912.5 LPZ011 1740.9 2073.5 1436.2
1075.8 4631.4 5232.7 4563.3 1080.6 5456.6 LPZ012 424.4 217.8 50.1
271.8 2286.7 713.5 1791.4 390.6 3209.9 LPZ013 395.4 123.8 33.1
181.1 3456.8 2121 2898.4 428.9 2673.3 LPZ015 490.7 210.7 60.8 130.8
2889.2 330.5 2123.4 230.2 2680.6 LPZ016 2411.9 710.9 346.5 1201.2
6897.9 4057.6 13340.1 3246.1 16664.2 LPZ017 635 257.4 36.2 247.2
2797.3 1219.1 3508 558.7 3953.8 LPZ018 1405.4 474.9 214.9 3188.7
8143.7 4992.8 12908.9 4208.2 14318.9 LPZ019 3487 975.9 698.4 8916.5
23680.5 15131 31656.3 14191 45343.3 LPZ020 251.9 195.6 0 40.9
1448.5 860.4 1113 305.5 971.2 LPZ022 250.1 112.4 0 133 2085.9
1282.5 2538.5 532 613.2 LPZ023 355.5 122.6 22.9 47.8 224 879.4
1419.1 132.3 605.4 LPZ024 366.5 108.9 45.7 94.8 225.3 723.1 1276.8
81 319.5 LPZ025 705.4 278.6 202.1 716.7 5145.5 4563.4 7215.5 1581.9
4195.8 LPZ026 268 100.6 9 92.1 1164.5 750.4 3973.6 275.5 642.3
LPZ028 386 174.2 96.8 374.9 4188.4 2733.8 7024.6 1186.8 2375.6
LPZ029 221.9 86.8 0 47.8 225.3 264.4 2172 162.4 957 LPZ030 319 166
67.6 146.9 801.6 1385.4 2283.4 145.8 1189.4 LPZ031 2010.6 881.7
538.1 754.3 4625.8 3395 4051.7 1327.9 5202.1 LPZ032 36097.5 35972.1
13659.1 19975.4 45544.5 20035.8 63759.8 31117.4 78268 LPZ033 1813.8
433.6 243.7 1171 7402.4 2278.7 10670.5 2204 5643.8 LPZ034 332.6 97
34.6 181 1097.2 0 2704.1 262.6 3481.4 LPZ035 248 60 0 71.4 1188.8 0
1332.5 153.6 3028.4 LPZ037 375.8 133.8 0 114.6 3024.3 909.1 2350.6
248.2 3546.8 LPZ038 577.7 237 44.3 370.7 4484.6 3572.5 4266 907.9
4571.4 LPZ039 965.6 406.7 361.2 537.9 4020 2304.1 4269.9 816 3717.5
LPZ040 399.9 127.4 0 88.3 1200.5 365 2123.2 244.3 1955.4 LPZ041
318.4 105.1 0 136.6 856.3 716.6 1528.5 245.6 1538.5 LPZ042 289.3
77.6 0.9 189.5 441.5 365.4 1007.7 239.2 1212 LPZ043 417.3 166.7
57.4 158.2 1197.2 1617.9 793.5 569.1 2018.7 LPZ045 754.8 310.3 152
691.5 4810.4 3305.3 4043.7 1476.3 3925.7 LPZ047 270.5 155.4 53.1
39.1 2165.6 579.6 980.9 361.7 1036.3 LPZ049 809.6 381.9 0 461.4
4406.4 2277.4 4764.6 2257.8 5528 LPZ051 333.1 121.5 0 56.6 1597.8
1677.8 891 270.7 1134.6 LPZ053 271 119.7 0 16 1662.4 2447.8 1202.4
201.1 827.1 LPZ054 345.4 131 61.7 79 1181.1 2238.5 1426.5 156.5
627.5 LPZ055 291 78.1 102.9 63.5 551.3 2343.8 1433.5 193.6 814.2
LPZ056 364.6 167.9 83.6 130.3 1816.9 2580.4 2589.4 343 1579.8
LPZ057 250 76 0 11.9 426 457.6 2589.6 113.4 709.8 LPZ058 231.1 40.8
6.2 44.4 454.7 163.4 3403.4 208.5 1300.4 LPZ059 239.2 78.6 0 15.7
189.7 267 3272.7 141 1439.2 LPZ060 235.1 26.7 29.4 35.6 524.8
1238.2 2231.2 182.5 1908.8 LPZ061 402.1 268.4 141.6 254.6 1694.1
3088 2343.5 557.3 4157.7 LPZ062 727.7 146.8 0 203.3 2873.3 2418.9
3109.6 926.6 5812 LPZ063 316.7 108.2 25 190 1837.3 1025.1 2727.2
421.8 4195.9 LPZ065 512.5 59.9 94.9 583.5 5694.4 3741.6 5366 1221.6
3911 LPZ066 622.2 66.8 23.8 405.6 6932.1 3089.5 4804.9 642.2 4079.9
LPZ067 883.7 218.7 0 358.7 4518.2 2342.9 5200.8 1318.4 5539.6
LPZ069 335.6 100.2 0 46.8 0 0.7 1736.5 255.4 2910.4 LPZ070 422.6
143.3 49.6 168.8 1231.5 1170.6 2432.2 262.7 2122.5 LPZ071 356 71.9
0 299.7 2303.6 1909.8 2575.9 447 2827.6 LPZ072 206.5 32.6 0 71.7 0
0 1362 189.6 907 LPZ073 374.2 129.6 32.1 125.8 307.4 691.8 1617.6
282.4 1306.9 LPZ074 434.7 86.5 0 76.6 1671.1 0 1112.5 232.7 663.5
LPZ075 298.5 173.5 0 53.5 4083.2 0 1143.9 113.7 335.1 LPZ076 209.9
83.1 7.2 7.4 2185.9 0 490.5 226.6 362.9 LPZ077 813.4 558.3 0 339.1
3733.4 4279.8 1115.9 671.7 4136.8 LPZ078 532 349.8 0 265.8 4460.1
3290.7 2776.8 686.3 3283.3 LPZ079 347.8 183.6 11.6 53.3 2115.4
1191.7 1451.2 160.5 1233.2 LPZ080 948.1 264.9 178 455.1 4633.9
2869.1 4230.5 1378.5 5566.5 LPZ081 313.7 96.1 100.2 59.7 1161.6
1470.2 1119.7 98.5 710.9 LPZ082 260.9 120.3 18.4 68.3 1734.5 1430.6
1581.6 160.1 379.4 LPZ083 284.9 134.6 12.6 7.8 1067 906.1 1482.7
107.2 790.5 LPZ084 1493.3 437.1 0 788 4497.3 4608.3 5078.9 2267.8
5087.7 LPZ085 468.1 139.4 0 62.1 1618.8 1355.9 4314.9 340.5 2421.5
LPZ086 601.3 91.6 11.9 332.9 3540.3 3354.6 6675.2 781.2 5545 LPZ089
457.1 124.1 26.2 267.7 2378.1 3364.6 3390.2 448.7 2930.7 LPZ090
436.6 50.7 0 149.1 3135.3 2087.8 3090.4 483.8 3408.1 LPZ091 350.2
91 96.2 99.2 1524.3 1976.3 3215 315.9 4532.8 LPZ092 387.6 33.6 0
114.4 2386.7 2600.9 2845.7 274.6 2036.7 LPZ093 195.6 25.8 0 27.1
2343.1 321.7 2875.2 252.7 1428.2 LPZ094 274.8 42.8 0 35.2 886.9 0
2502 152.6 1571 LPZ095 288.6 91.5 0 14.1 3.5 0 2317.2 144.4 1427.4
LPZ096 475.4 138.7 13.1 199.2 1690.8 1415.2 3939.2 207.2 1889.7
LPZ099 428.1 73.8 0 185 2531.8 2596.6 3925.9 399.8 1967.5 LPZ100
474.3 131.1 24.6 266.6 2710.6 2276.7 3358.6 503.5 2245.1 LPZ101
492.1 101 81.9 233 2341.4 2076.1 2199.9 423.1 2022.4 LPZ102 477.3
133.2 0 167 3722.9 1604.4 2432.1 386.5 1199.8 LPZ103 353.5 90.7 0
108.9 4756.8 214.4 1447.5 203.9 760.7 LPZ106 534 199.4 58.6 117.6
3052.4 1259.2 1610 277.6 1224.3 LPZ107 29718.3 56194.4 31132.5
35651.5 44972.5 20589.6 38396.3 32828.7 75965.1 LPZ108 852.4 433.1
161.4 417.4 4354.8 2259.1 3536.9 1168.5 3161.8 LPZ109 554.2 248.6
83.8 254.6 3303.4 2521.1 1972.8 608.5 2858.5 LPZ110 614 203.3 166.1
182.1 3236.7 2792.6 2764.1 455.5 3139.2 LPZ111 349.5 167.7 77.4
82.9 1407 1386.1 1469 200.9 1397.5 LPZ112 497.3 279.2 65.3 242.6
3553.5 2504.1 2454.6 328.3 2182.2 LPZ114 890.2 346.7 0 399.3 4520
3367.9 2902.7 1229.6 3474.1 LPZ115 24782.8 12016.1 1401.9 12188
32718.2 17087 38203.9 17191.1 25318.5 LPZ116 1388.1 392.7 0 884.6
8895.4 3131.2 15554.1 2195.7 5401.6 LPZ117 6228.4 4810.2 389.7
1298.8 4199 2671 5473.5 1488.2 3911 LPZ118 424.5 267.3 39.9 196.2
2507.4 2210.1 2856.3 370.3 2640.4 LPZ119 295.1 183.4 0 59.8 2443.3
1153.2 2040 158.7 1132.1 LPZ120 213.5 88.4 49.2 185.9 1336.6 1390.4
1604.5 186.7 1360.8 LPZ122 317.6 120.3 0 92.1 1390.2 2045.1 1701.2
112.5 1307.5 LPZ124 346.6 173.2 38.9 119.4 2097.9 543.9 2679.2 221
1941.7 LPZ126 436.1 185.4 0 142 1762.2 0 2164 283.2 2343.6 LPZ127
455.7 208.6 0.1 109.8 1091.7 1925.5 3143.4 345.4 2565.8 LPZ128 421
602.3 25.1 651.1 4496.3 2712.3 5456.8 1177.7 3875.7 LPZ131 287.1
401.2 0 58 1931.2 422.4 3510.2 232.2 1275.8 LPZ133 377.8 399.9
141.9 139.5 2396.5 1989.9 4287.5 388.9 1119.6 LPZ136 455.2 191.7
103.3 239.9 2289.7 1775 2586.1 322.4 1429.8 LPZ137 398.1 123.1 0
214.7 2480.2 1118.7 2138.5 411.3 1935.9 LPZ138 1987.8 1102.2 11.9
1112.7 4785.6 4242.8 3044.7 1298.3 2786.7 LPZ140 401.4 205.9 115
316.7 2737.2 1950.6 1491.6 414.2 2031.4 LPZ141 917 621.2 0 726
4938.4 3889.9 3237.4 1109 4165.8 LPZ143 4702.9 1483.9 774.8 6791.2
20187.4 6913.4 23175.5 10795.1 26322.7 LPZ144 529.7 392.6 25.7
303.5 2644.9 1902.4 2380.9 486 2422.7 LPZ145 410.2 206.5 25.4 152.3
1618 2219.1 2012.8 357.8 2494.3 LPZ146 294.2 152.3 0 125.3 1063.5
1142.1 1129 215.1 1927.1 LPZ147 366.5 238 0 246.5 2120.4 956.1
1343.4 260.2 1539.3 LPZ148 390.6 212.9 0 170 2107.1 2195.4 1832.6
1240.9 3839.8 LPZ149 1872.8 1139.7 218.5 1748 6204.2 1641.3 5144.7
3526.5 5193.6 LPZ150 1958.1 1284.1 645.6 10615.2 29419.9 2976.7
52052.2 21478.7 41125.9 LPZ151 3477.3 1936.4 155.4 1423.9 5253.1
1925.1 5434.9 2248.3 5180.5 LPZ152 963.2 481.9 42.2 658.9 4770.8
3607.7 4661.8 1003 4661.9 LPZ153 13685.9 27883.7 11205.9 13827.1
24872.9 15412 26204.7 12163.3 41713.9 LPZ154 621.9 470.5 45.6 381.1
2965 2584.3 2802.2 416.8
2738.4 LPZ155 2004.5 1513.1 388.4 1358.9 4265.3 4159.3 4386.4
1022.6 4543.9 LPZ157 2978.4 1332.8 407.8 1400.6 4650.6 3692.3
4760.8 1195.9 5002.5 LPZ158 12352.4 18933.2 12155.7 9376.4 23120
15280.6 18384 10293.8 50995.4 LPZ162 3778.2 4069.3 426.4 2285.5
6458.4 3004.6 5794.3 4161.4 13171.9 LPZ165 1181 805.1 0 756.4
4641.9 2702.4 5464.6 1470 4822.6 LPZ166 4624.9 5016.4 0 2508.8
9907.8 1939.3 5655.3 2999.9 5667.6 LPZ167 3339 1655.9 319.8 1101.8
4807 4980 4678.8 1968.2 4248.1 LPZ169 787.5 556.1 226.4 461.9
2830.2 2225.4 3549.7 684.4 3249 LPZ170 851.1 501.5 0 589.6 4405.2
4440.6 4652.4 1622.7 4556.9 LPZ171 1325.6 612.2 0 697.6 3647.5
3148.7 3446.8 1053.8 4043.4 LPZ172 748.6 490.4 0 802.4 3953.2
2939.1 3550.9 848.3 3809.5 LPZ173 4460.6 3415.7 1050.1 5769.8
16078.4 6658.8 16861.6 8623.4 21007.1 LPZ174 501.9 308.6 36.2 358.6
3054.1 1731.2 1954.9 862.9 2773.9 LPZ175 1476.5 1057.1 181.6 836.9
3731.4 3120.9 3879.4 755.9 3687.2 LPZ177 302.3 228.3 18.6 107.1
753.6 1430.4 941 165.4 1411.2 LPZ179 616.8 314.9 8 379.4 4544.7
2954.4 3425.3 1361.5 5310.3 LPZ181 1103 430.7 0 671.3 4917.1 3821.3
4976.3 2646.1 5785.8 LPZ182 992.8 435.1 0 468.1 3930.1 4177.6
3287.5 909.9 4820.2 LPZ186 2455.7 1428.7 760 2414.4 9679 4431.1
5537.9 3859.3 5921.7 LPZ189 40770.3 68311.4 75133.7 45673 47303.1
21072.6 66542.7 34849.1 78485.4 LPZ194 612.8 572.1 155.6 376.3
3673.5 2240.2 3365.2 469.5 3608.6 LPZ195 676.9 346.6 32.5 459.9
4278.1 4622.4 4442 1006.8 4754.3 LPZ196 2923.1 1787.1 448.6 1537.1
4490.3 3684 4875.6 1426.9 4447.8 LPZ197 592.3 177.5 343.1 629.4
5825.4 3639.7 4308.7 649.9 2838.2 LPZ198 801.7 295 8.2 511.1 4384
3156.4 4823.8 1125.6 3947.2 LPZ199 402.5 165.1 62.7 265.9 2158.3
2363.7 3531.7 423 2360.1 LPZ201 1338.5 361.2 209.6 1053.7 4997.8
4565.6 5107.6 2350.9 5882.2 LPZ202 8178.9 2795.3 2359.9 12950.9
39890.9 20247.9 62085.4 31826.4 78541.6 LPZ203 1983.5 1083.8 1146.8
2044.1 9173.9 2174.8 11135.5 4073.8 19571.3 LPZ204 38154.8 66241.1
88440.7 45372.1 47166 21054.3 65826.6 33875.6 78581.7 LPZ205 1079.5
521.7 589.1 758.4 4742.5 3837.7 5220.2 1453.2 4888.6 LPZ206 947.6
700.1 505.1 697.8 4089.6 2774.1 5242.9 1190.3 4578.7 LPZ207 39899.6
60813.3 95637.5 45997 47123.6 21073.2 63996.4 35149.4 73080.5
LPZ208 27268.6 27230.4 39428.3 29579 40216.5 20480.2 42465.8
26526.9 76872.4 LPZ210 683.8 538 429.6 439.8 2402.5 3285.4 2419.1
636 3965.6 LPZ211 603.1 530.3 87.2 434.6 3653.3 2234.5 1744.1 359.8
1644.7 LPZ212 1018.4 580.7 155.2 1734.1 9212.3 2338.9 5061.9 2257.3
4303.3 LPZ213 465.6 327.1 33.1 296.6 3012.3 2419.4 2198.4 570.9
2013.9 LPZ214 245.6 267.4 0 160.5 912.5 756.5 880.7 133.9 936.8
LPZ215 981.1 492.8 52.3 828.3 5390.6 4771.4 4868.1 2893.1 15622.6
LPZ216 9410.5 3834.8 1843.2 14492.9 43137.1 21097.9 63778.6 34830.7
78538.8 LPZ217 31119.9 47126.3 30689.1 28515.4 38695.3 19884.5
37913.9 21282.9 69590.2 LPZ219 1300.2 886.5 475.3 946.3 4898.3
4361.6 3705.4 1784.6 5810 LPZ220 5695.5 7233.6 9375 4579.9 6540.8
4340.1 5612.6 2993.6 5852.3 LPZ221 242.6 209.5 0 186.3 2553.4
2132.5 1632.9 258.2 3202.9 LPZ222 186.9 130.5 0 163.4 2138.4 1386.3
1855.4 223.3 2407.6 LPZ223 289.9 168.4 0 164 2019 1582.7 2374.4
250.3 2309.2 LPZ224 308.3 114.3 20.5 333.9 2467.7 2535.2 2722.9
518.9 3054.9 LPZ225 5540.8 1287.2 919.3 7087.6 18926.6 9277.7
20292.9 8705.5 19974.7 LPZ226 179.1 111 9 136.8 1049.8 2071.1
3332.8 463.4 2517.8 LPZ227 243.5 136.2 0 341.4 2502.5 2597.6 4869.5
3226.2 16356.7 LPZ228 470 249.6 0 349.2 4062.6 3388.6 5688.3 2586.2
15112.1 LPZ231 480.4 296.3 87.4 326.6 3832.4 2343.1 5557.3 1542.5
5679.2 LPZ233 468.6 332.9 161.7 350.3 2519.3 1967.1 5201.8 996.1
4833.8 LPZ234 525.8 368.6 287.8 380.3 1626.6 1285.8 4231.8 1235
5329.5 LPZ235 310.7 233.3 0 370.2 1746.3 1474.1 4135.8 541.1 3382.9
LPZ237 683.5 468.8 487.2 506.9 3683.2 2108.7 3966.8 1369 5254.5
LP2239 546 192.6 339.2 331.5 2584.8 2384.8 3215.7 754.4 4041.1
LPZ240 353.7 280.8 163.9 274.8 2281.8 1696.4 2345.8 319.2 2180.8
LP2241 283.3 260.2 0 206.4 2481.1 1787.8 775.1 426 1729.3 LP2242
7478.4 2664.4 1039.1 7024.1 22393.3 11120.5 20928.9 8581.6 22951.9
LPZ243 242.4 167.1 0 157.2 2175.8 182.4 806.9 237.7 977.2 LPZ244
350.3 206.1 0 409.4 4522.7 3997.1 4486.3 1022.8 3387.1 LPZ246 260.9
200.9 0 251.8 1930.9 1335.1 995.1 1033.7 4712.9 LPZ247 438.3 274.4
0 341.4 2994 2325.3 2926.7 1508.7 5494.3 LPZ248 748.4 714.1 291.3
878.4 3814.2 2737.7 3489 1155.1 4689.5 LPZ249 373.3 375.6 0 613.1
4798.1 2088 3560.8 433.5 2486.3 LPZ250 159.5 201.7 0 317.6 2037.8
2085.9 1721.8 220.5 2051.7 LPZ251 141.7 157.9 0 178.9 1377.4 1723.8
1136.8 91 1271.1 LPZ255 220.8 176.1 0 646.5 4160.8 2725.5 5110.3
1217.2 5217.7 LPZ256 94.6 101.7 0 149.5 821 812.6 1989.5 65.4 432.2
LPZ257 147.9 118.4 0 135.1 1206.9 1208.1 2197.2 207.9 745.8 LPZ258
168.3 124 0 174.7 2264 2172.4 2672.8 532.8 2245.2 LPZ260 213.5
172.9 0 141.3 1172 2974.6 4118.3 232.4 1057.4 LPZ261 147.2 78.5 0
126.6 1212.5 2349.9 4604.9 139.4 741.6 LPZ264 318.3 174.1 0 201.7
3104.4 2387.6 5505.8 582.7 3775.1 LPZ265 1566.8 449.9 129.7 646
4992.3 3477.7 5635.5 988.7 4307.2 LPZ266 92.8 287.2 0 132.5 931.4 0
4689 190 726.3 LPZ268 171.3 217.2 0 206.8 2142 2135.8 5156.3 296.8
1912 LPZ269 1530.1 571.5 419.3 2333.1 11130.7 4947.4 13881.5 5155.2
5755.6 LPZ270 162.3 291.4 0 450.4 3822 3736.4 4342.8 978.4 2987
LPZ271 454.6 266.6 45.9 381.8 3194.7 2859.7 3598.7 1277.5 4054
LPZ272 2943.2 763.9 613.2 1451.3 7894.9 2900.8 5222.6 3222.4
16317.5 LPZ273 215.5 178.7 0 112.2 1288.3 908.3 145.5 182.5 544.8
LPZ274 271.5 189.3 0 322.8 3311.7 1141.5 1301.4 620.8 3182.2 LPZ275
174.5 152.8 0 99.6 1052.3 0 0 109.3 887.1 LPZ276 146.8 139 0 129.4
1165.3 123.5 505.4 82.6 461.9 LPZ277 201.8 137.5 1.8 57.1 761.5
931.9 497.5 106.1 1427.3 LPZ278 177.9 152 0 76.6 588.2 1424 311.2
107.3 1178.8 LPZ279 183.3 179.3 0 304.9 2458 1032 524.7 276 1530.9
LPZ280 142.1 125.5 0 125.6 1116.5 623.5 1147.9 125.4 770.5 LPZ281
18 109.8 0 58.9 563.4 850.5 564.6 11.5 317.8 LPZ282 54.3 164.1 48
95.5 1493.6 1874.9 1033.3 54.7 844 LPZ283 1607.8 392.6 48.4 1220
6358.1 2922.6 5552.8 1970.3 5032.3 LPZ284 42.5 119.4 0.3 48 804
748.4 1365.8 66.6 0 LPZ286 34.3 164.5 0 74.8 973.6 1463.4 1205.7
81.2 329.3 LPZ287 118.8 186.4 0 116.4 1573.7 1568.6 2124 252.9
884.4 LPZ288 103.4 162.8 0 78.3 1328.1 2890.4 4192.3 196.9 598.1
LP2289 137.1 87 0 113.6 2096 2101 5147.9 332.3 913.7 LPZ290 1598
425.5 186 1782.8 10543.9 4598.5 16706.4 3923.8 5648.6 LPZ293 155.8
225.7 0 129.9 3228.2 2291.5 4891.2 316.9 2001 LPZ294 65.5 180.6 0
66.6 2237.5 527.7 4397.4 127.5 688.2 LPZ295 119.9 258.8 0 149.7
1964.6 560.6 5062.1 141.5 897.3 LPZ297 333.9 277.6 59.4 830 5667.2
3950.7 5680.5 1452.6 3614 LPZ299 102.6 225 12.2 268.1 734.7 898.2
4025.5 371.2 913 LPZ300 231 271.7 97.1 42.6 113.2 1713.7 3264.5
4295.2 631.1 LPZ301 272.7 378.2 155.6 97.2 77.2 2110.4 1733.8 485.6
1392.8 LPZ303 145.6 184.3 641.8 52.8 1562.5 1072.8 365 55.5 358.3
LPZ304 422.5 346.5 108.2 350.8 3262.7 2215 1102.8 264.1 1534.7
LPZ306 2207.6 484.7 609 3182.2 9671.4 3639.8 17157.3 3556.4 15015.7
LPZ307 1761.1 1119.4 454.7 846.1 4114.9 2673.3 4082.5 554.8 2983.5
LPZ308 153.5 213.5 85.5 113.3 1369.9 1433.3 133.8 123.2 1146 LPZ309
132 192 14.3 49.2 1137.6 1626.7 126.9 85.1 805.1 LPZ310 325.9 353.6
311.3 253.1 4210.4 2703.3 1846.5 513.3 3102.9 LPZ311 176.9 217.7
72.3 245 3652.2 4352.6 4175.7 734.7 4625.7 LPZ312 70.4 177.2 139.4
53.2 2094.2 1461.1 945.5 57 274 LPZ314 247.5 221.2 217.2 66.9 1767
753.8 872.4 93.1 751.2 LPZ315 167.6 220.4 322.5 172.9 3442.4 1985.7
2505.1 698.5 3667.7 LPZ318 912.5 297.8 441.9 957.6 7473.1 2682.6
6826.4 2471.8 5148.9 LPZ320 7.3 212.8 154.8 55.8 1682.1 1548.5
1038.5 111.1 822.5 LPZ321 199 259.6 157.2 96.8 2588.8 2465.4 3436.8
409.2 3989 LPS001 369.9 656.8 1322 4095.5 4733.4 7892.2 3248.2
5064.2 6260.9 LPS003 442.5 392.9 262.1 648.7 2035.7 570.6 4524.4
1332.8 543.9 LPS004 811.4 552.2 515.2 1694.9 2438.2 425.1 1218.8
714.3 86.8 LPS006 271 141.2 200.2 0 662.1 1245.1 261.5 1282.6 405.7
LPS007 212.1 91.8 139 47.7 243.4 0 929.2 568 267.1 LPS008 197.6
196.8 224.7 0 235.1 2144.5 1034.5 985.2 307.3 LPS010 152.7 133.4
253.8 241 299.5 1667.7 242.1 1218.8 1170 LPS011 186 136.1 257.9
816.5 257.4 2372.2 77.7 1377.7 1346.8 LPS012 317.2 238.4 236.4
817.5 1316.7 1626.1 0 1324.1 564.7 LPS013 349.5 403.6 412.9 1840
1582.6 2769.8 215.4 1196 1328.8 LPS014 511.4 1574.1 257.3 4169.4
5035.7 4543.5 1016.4 3744.8 3779.2 LPS015 2495.9 3076.7 1720.8
7899.3 3640.5 7103 1228.1 5376.7 11989 LPS019 1028.2 604.5 239.7
3074 2189.3 2161.1 1239.9 2955 2426.7 LPS020 324.1 222.9 120.5
1214.3 1056.8 676.2 872.7 916.8 1058.1 LPS023 160.9 116.3 50.3
623.9 1984.1 0 640.7 349.7 694.3 LPS024 359.3 324.3 198.9 1596
2789.8 1253.1 1212.2 2644.1 3158.3 LPS025 614.7 616.8 493.6 3452.4
3736.3 3726.5 1796.1 4133.4 5569.5 LPS026 153.7 494.7 0 3053.4
3077.5 2157.2 806.2 3886.5 2133.1 LPS027 132.2 267.1 0 1309.2
2323.4 1330.2 1501.6 1931.3 1355.1 LPS028 214.3 446.5 155.7 2472.5
3336.1 2467.9 2987.9 4507.5 3530.8 LPS029 202.7 384.3 223 2040.4
2060.5 2600.8 7214.8 4150 3867.7 LPS030 113.5 132.5 0 515.6 2793.9
110 2158.7 2684.7 1237 LPS031 168.7 123.5 11 557.6 3086 812.5
6212.6 2534.1 3290.9 LPS032 145.2 160.9 1.7 650.5 2144.2 1070.8
1474.2 2819 3561.4 LPS036 582.6 616.4 200.9 2224.2 2656.4 1723.6
3073.1 1866.1 2955.4 LPS037 502.6 620.4 219 1626.6 3359.2 2705.7
2125.3 2456.9 2004.1 LPS038 962.6 216.5 375.7 0 1256.5 930.9 1492.9
1578.3 1406.9 LPS040 228.9 86.1 158.6 0 256.8 0 245.1 758.3 1.7
LPS041 222.7 149.2 123.3 0 252.4 0 965.1 1065.9 553.4 LPS042 447
661.1 489.9 1672.8 2520.7 492 1046.8 2834.7 1940 LPS043 333.7 264.6
407.3 656.8 1046.4 546.5 1412.3 1097.9 948.4 LPS044 249.8 277.8 652
1327.3 907.5 1110.5 1892.3 1353.4 1674.8 LPS045 250.7 107.2 177.9
302.6 231.9 944.7 1881.7 0 485.4 LPS046 232.4 285.6 224.5 1302.1
1872.6 1104.6 2610.7 1128 1744.2 LPS047 2649.2 6969.7 2792.6
14436.1 10141 5428.8 4057.3 2999.2 13647.7 LPS050 2428 7502 3442.1
12204.8 7385.4 9850.6 3395 8052.4 14726 LPS051 478.2 219.7 175.1
1861 2222.1 1876.9 1922.5 800.9 2052.9 LPS052 328.2 196.2 138.3
984.6 1458.3 1324.4 1585 1112.9 1726 LPS053 264.2 111 0 1370.2
1524.9 1548 2692.1 2941.5 3096.1 LPS054 285.4 231.1 8.5 1370.4 2138
1273.8 2930.8 2762.2 2604.2 LPS055 1192.2 1118.8 180.5 5840.8
4203.1 3690.8 1865.1 3205.9 4824.3 LPS056 1541.6 2959.9 1354 9519.2
8084 6331.3 1820.6 5902.8 11668.9 LPS057 214.5 406.9 0 2028.8
2744.4 756.5 2136.1 1844.8 2454.6 LPS058 277.4 244.5 70.8 1623.9
1826.1 1626.5 2708 2514.4 4077 LPS059 115.3 135.2 1.5 996.9 1194.7
1022.8 1723.1 1265.3 2390.7 LPS060 163.3 268.5 0 1866.5 1707.3
1953.7 2184.7 2422.6 2990 LPS061 222.7 255.6 53.4 1448.5 2146.6
1600.7 1956.2 2511.7 3332.5 LPS062 136 228.5 99.1 627.1 863.2 467.7
1610.3 2304.9 2842.2 LPS063 299.7 251.8 226.3 796.8 1427 1771.5
1174.1 930 1433.9 LPS064 3079.8 4014.9 3039.3 7349.2 10807.8 7372.1
10515.8 4426.8 13038.7 LPS065 434.1 644.6 313.7 1147 1456.3 3097.5
2632.9 3695.2 1575.1 LPS066 214.7 171.7 134.2 0 69.3 249.7 726.1
871.9 586.6 LPS067 706.6 686.2 488.8 3013.5 2498.9 4522.1 4844.3
4782.1 5775 LPS069 199.6 172.1 123.9 75.4 19.4 269 874.1 854.6 0
LPS070 143.6 186.9 117.9 289.4 685.5 222.6 528.3 582.6 322.6 LPS071
180.4 170.2 187.2 157.2 183 882.3 326.7 508.4 310.2 LPS072 235.9
170.8 169.6 449.9 290.5 777.5 456.3 283.3 479.2 LPS073 900.4 1318.7
629.3 3416.1 4420.4 3894 4010.1 3367.3 4106.8 LPS074 27858.2
33812.3 32162.2 44513.2 111430.2 87262.8 47575.8 18233.4 66903.6
LPS075 2119.6 3296.9 1347.3 9540.3 5518.1 6367.8 10437.2 4054.1
9821.4 LPS076 347.6 336.3 218.5 2343.7 2326.5 1569 2415.2 1580.7
1990.4 LPS077 568.1 612.4 550.1 2908.9 1727.9 1660.2 2164.5 1798.6
2588.1 LPS078 3174.9 3137.6 3222.6 7616.1 6945.1 9024.5 11397.6
7995.6 26362.3 LPS079 1049 1066.4 302.2 4400.8 4126.8 4404.7 8203.7
4645.6 9377 LPS080 21208.7 28180.7 9065.4 39068.1 63741.1 37523.8
35948.5 11444.8 57266.6 LPS081 27381 33419.5 10292.3 43529.2
63629.5 28119 42128.9 15984.6 62043.4 LPS083 711.4 825 64.8 3306.9
3733.1 1898.2 3688.2 3407.5 3959.9 LPS084 216.4 211.2 21 1350.5
1724.2 1394.5 1965.6 2089.4 3604.5 LPS086 185.6 214.1 0 1808.2 1476
2915.6 2342.5 932.2 3339.1 LPS087 3404.7 5840.3 4144.1 12101.2
12860.9 14601.5 24953 5018.8 19643.5 LPS088 165.2 224.6 62.5 1497.7
2813.8 1593.8 3740.9 4017.3 3934.9 LPS089 223.8 213.7 0 1318.9
1574.5 2141.4 2443.5 3799.4 4185.6 LPS090 398.7 693.2 142.2 2593.7
2695.2 3465 3755.9 3638.7 3587.8 LPS091 391.2 700.6 270.7 1469.2
2092.2 3047.2 3754.2 3524.1 3149.7 LPS092 286.1 376.1 254.9 235.2
433.1 1353.8 1747.8 2658.1 3246.6 LPS093 185.6 273.7 126.3 114.7
296.8 254.7 412.1 1076.9 483.7 LPS094 232.8 261.8 151.7 274.3 249.9
641.1 891 577 1102.1 LPS095 199.3 191.7 91.4 25.7 169.9 433.7 813.4
1394.3 807.7 LPS096 97.9 139.2 63.2 0 162.4 159.2 504.6 752.3 150.6
LPZ001 150.8 207.8 257.7 202.6 485.5 704.2 555.2 1924.3 528.5
LPZ002 154.6 167.7 323.6 937.4 717.4 755.6 1068.8 1089.8 1107.7
LPZ003 1348.1 1781.1 609.3 5019 4261.6 6148.5 5600.4 3419.6 4858.4
LPZ004 2798.4 5533.1 2921.8 10703.8 8020.5 9958.4 10424.8 4135.9
14742.8 LPZ005 8403.4 19547.7 8589.6 32699.4 31426.9 21580.2
19437.9 8059.6 14660.3 LPZ006 360.6 2106.1 1173.8 7812.9 7966
11310.2 10516.8 4661.6 4618.2 LPZ007 272.5 409.9 454.7 2038.3
1107.6 2043.7 2073.5 2249.9 2751.3 LPZ008 207.7 258.2 212 1939.1
1482.6 1926.1 2243.5 1036 3324.9 LPZ009 745.1 496.5 633.9 4276.8
7474.5 9130.4 9814 5721.3 14116.8 LPZ010 893.7 1464.1 326.6 3759.5
4034 4261 4672.2 4388.6 9625.3 LPZ011 1829.5 2488.7 350.4 5922.7
4285.4 2984.2 5579.8 4236 9972.7 LPZ012 227.8 289.9 28.6 1885.9
1660.5 843.6 1913 1434.5 2785.5 LPZ013 247.7 213.8 84 1553 2015.3
1547.3 2567.6 3196.5 4347.7 LPZ015 261.7 315.8 55.6 2254.3 2409.8
2190.1 2562.4 1291.2 3237.3 LPZ016 2750.3 2151.8 3003.8 8316.2
6689.1 9147.4 9444.8 3349.6 8167.8 LPZ017 582.2 701.3 227.4 3830.1
3650.3 3828.2 4552.3 4574.5 4476.8 LPZ018 2867.6 6184.2 2746.8
10513.4 9443.1 10880.7 12748.3 5491.2 19422 LPZ019 7551.3 15875.3
9232.8 20440.4 22870.4 31026.1 33842 12823.6 38075.4 LPZ020 293.1
896.2 143.1 1661.8 2519.9 2987.1 4132.4 4022 3145.1 LPZ022 213.3
493.5 173.5 82.5 467.8 1355.1 1041.1 1481.9 1035.5 LPZ023 191.2
616.7 118.1 78.4 184 955.2 516.3 1254.6 574.4 LPZ024 142.6 321.7
118 0 81.9 826.8 195.8 763.9 491.7 LPZ025 661.9 764.4 536.9 1885.6
1791.8 3200.6 3017.8 4028.4 3897.8 LPZ026 194.7 221 150.4 1102
513.6 1714.4 1291.2 1625.6 980.7 LPZ028 301.3 424.2 210.9 1467.7
1654.3 2848.4 1937.1 4092.7 3086.2 LPZ029 132 151 124.8 319.5 644.1
478.7 452.4 564.8 709.3 LPZ030 305.3 616.6 170.6 2800.7 2572 2485.4
1960.4 1425.4 2381.1 LPZ031 1945 3098.6 3636.7 12422.3 7673.2
8643.5 11552.7 4295.4 4600 LPZ032 26761.5 33518.4 33623.2 45482.1
106536.1 114284.5 46968.1 16371.6 42282.1 LPZ033 2068.3 1779.5
6651.8 7887.8 5249.8 9848.7 7632.1 4500.7 7710 LPZ034 221.4 363.3
216.3 2503 1949.2 1674 2078.9 1428.3 1774.9 LPZ035 110.7 156.8 85.2
836.6 512.1 1355.7 1217.4 294 1525.3 LPZ037 229.5 206.2 186.5
1422.1 1962.7 2742.9 3023 614.1 2895.5 LPZ038 605.7 722.8 352.5
3551.8 3072.2 3614.2 3266 2494.6 4039.7 LPZ039 366.4 964.6 177.1
3755.2 2744.9 4599.4 3589.7 2407.8 3925.2 LPZ040 185.6 278.3 100.8
2131.2 1321 1479.7 1654.6 773.7 2087 LPZ041 119.9 120.8 5 1199.8
1220.8 1090 1431.2 630 2206.1 LPZ042 61.9 121.9 2.6 731.4 1897.5
986.4 1366.1 458.4 2625 LPZ043 357.9 355.1 0 3236.5 2746.8 2960.1
3138.2 911.1 3345.6 LPZ045 738.6 1003.7 565.3 3866 3168.1 6406.2
4028.7 4526.2 4573.4 LPZ047 139.7 133.1 0 481.3 857.9 831.8 954.1
1926.3 3129.6 LPZ049 1396.5 2125.5 1496.9 4514.4 3629.8 5942.4
6898.6 3610.7 9214.2 LPZ051 264.6 610.7 205.1 826.3 1819.9 2243.8
3000.9 3400.7 2810.3 LPZ053 174.9 827.9 152 161.2 563.8 1149.7
1277.9 1243 1383.5 LPZ054 205.7 951 128.4 976.1 1901.4 1626.6
1265.8 1437.6 1328.8 LPZ055 135.2 389 168.5 420.2 524.3 1650.6
848.4 1200.6 914.2 LPZ056 190.2 323.3 229.5 439.9 664.2 1613.1
1014.2 1727.3 1126.2 LPZ057 87 199.9 180.1 2154 863.7 3059.1 2994.9
2696.3 2990 LPZ058 139.3 227.5 55.3 1695.1 902.5 2426.6 2195.6
1925.4 1598.2 LPZ059 173.4 289.6 189.4 891.7 759.7 1835.3 1332.9
962.5 1286.6 LPZ060 301 464.8 114.1 2296.5 2860.4 2786.6 2974.4
1629.6 2301.9 LPZ061 1212.2 1711.1 794.4 7468.5 5190.1 7957.5
5857.5 2819.8 3922.3 LPZ062 2078.9 3648.3 2499.3 14932.7 7691.9
9294.1 9213.3 3077.8 4850.3 LPZ063 641.6 984 2114.2 5547.2 3688.6
6191.3 4844.5 4058.7 4162.4 LPZ065
520.2 443.3 332.3 2720.4 1816 2848.5 3320.4 4501 3948.4 LPZ066
663.7 357 356.6 3458.6 2196.3 3567.6 3081.4 1325 2388 LPZ067 1469.5
2582.6 3152.4 8674.9 8080.1 9367.6 8556.2 4135 7240 LPZ069 211.4
283.6 0 1921.6 913.1 1567.5 1866.5 1043.3 2269.1 LPZ070 229.6 334.9
2.8 1659.3 1254.6 1681.6 1883.6 1360.7 2442 LPZ071 332.3 633 15.7
3126 2729.9 3290.2 2998 2011.7 2744.8 LPZ072 39 38.9 0 581.2 1401.7
1307.1 1089.7 710.4 1866.3 LPZ073 131.3 250.9 4.4 1176.4 2903.2
2356.7 1718.3 985.9 2700 LPZ074 92 116.5 0 355.6 1643 1041.3 1027.4
1042 2553.6 LPZ075 195.9 0 0 0 474.7 488.1 847.6 1488.4 2755 LPZ076
232.4 134.5 730.7 0 268.9 0 568.3 1007.4 2624.4 LPZ077 1143.1
2350.7 187.5 6551.1 5960.9 5520.2 7189.6 3483.6 7931.5 LPZ078 851.5
1021 873.3 3011.7 4619.7 5273.6 6408.3 3950.1 8831 LPZ079 281.6
779.7 315.9 1296.6 2065 2090.8 2287 2271.9 1824.5 LPZ080 1653.5
3124.5 3778.7 8321.1 7987.7 10470.3 8085.7 4454.9 8067.4 LPZ081
92.6 292.7 161.8 746.8 903 1558.5 1410.8 746.1 1230 LPZ082 123.2
430.5 240.1 1907.9 1283.9 2707.5 1801.7 948.9 1634 LPZ083 77.8
183.2 120.2 4010 2437.2 4390.6 3649.7 3983.1 7032.5 LPZ084 1272.3
956.1 1297.2 5881 4505.3 10642.2 9798.6 3938.5 8446.3 LPZ085 321
466.2 457.6 4324.8 4008.4 6890.5 4599.7 5672 10082.2 LPZ086 1529.6
3587.2 3236.8 10729.4 10010.2 10739.2 10634.9 4674.2 11980.7 LPZ089
614.3 500.4 601.9 4196.2 3890.5 4405.8 4331.9 3066.9 3960.3 LPZ090
643.8 1177.5 1315 4017.8 4456.2 6394.5 4824.4 3000.1 3538.3 LPZ091
1006.7 1754.2 4090 11615.8 11728.5 16837.8 14461.9 5005.1 14726.4
LPZ092 419 528.6 1336.8 4158.4 3568.1 8393.4 8192 4638.4 3939.4
LPZ093 162.2 453 90.9 1436.5 899.2 2218.6 1798 741.8 1789.8 LPZ094
123.1 197.1 0 1355.7 779.4 1360.5 1713.8 794.4 1597.8 LPZ095 94
130.6 3.5 1099.9 737.4 558.3 1473.4 871.4 1809.3 LPZ096 314.3 327.5
22.1 2574.2 1620.9 2748.1 2533.9 1858.4 2979.9 LPZ099 231.7 359.8
6.2 1456.9 1335.7 1672.4 2170.8 2160.8 2755.8 LPZ100 375.6 650.4
136.7 2834 2518.6 3053.2 3159.8 2841.2 3453.7 LPZ101 217.1 425.7
11.8 2769.3 3312.6 2556.3 5262.5 1501.2 2947.3 LPZ102 294.3 289.2
55 1803.6 2764.5 2532.5 2613.4 2447.6 3377.6 LPZ103 224.2 92.8 17
989 1850.6 1643.7 2303.4 3729.5 4029.5 LPZ106 328.4 158 57.5 912.5
1239.6 1214.5 1669.6 2062.3 4720 LPZ107 25137.4 28865.6 19438.9
43316.7 75674.1 58296.4 45927 3332.1 64391.6 LPZ108 1132.8 2084.1
330.6 4299.2 3629.8 4480 6406.5 3235.2 9993.6 LPZ109 548.8 1356.1
417.7 3237.9 2581.9 3177.5 3977.1 1874.2 4107.7 LPZ110 379.7 1132.3
220 3941.9 2720.7 4103.6 3563 2245.7 3031 LPZ111 157.5 383.2 200.3
1188.3 749.8 1675.7 1926.1 1685.1 1655.3 LPZ112 332.1 522.9 220.7
2950.9 2223.9 2257.5 2856.4 2632.8 2711.8 LPZ114 590 151.8 217
4659.6 3824.5 6848.2 7228.9 2831.3 9446.9 LPZ115 6268.6 3368.3
8724.6 25695 19481.5 33011.8 32267.9 2903.4 52760.4 LPZ116 891.2
766.7 1481 6455.1 3684.7 5597.5 8456.7 1682.9 10139.2 LPZ117 529.8
826.6 126.5 3706.8 2663 2897.5 3996.1 1807.3 2635.2 LPZ118 437.8
628.1 122.1 3903.5 3245.2 3529.9 3500.1 3506.1 2867.9 LPZ119 331.3
755.1 83.1 3761.2 4072.6 4024.9 3449.9 3742.2 2533.9 LPZ120 196.5
796 181.4 4219.4 3297.7 4322 4539.3 2622.7 3763.6 LPZ122 154.4
208.4 158 2332.3 1468 3044 2838.8 2112.5 2323.6 LPZ124 172.9 420.1
136.6 2319.5 1539.3 1722.7 2453.6 3109.1 2297 LPZ126 446.5 604.2
187.3 3859.4 3120.8 2816.9 3324 2181.4 3237.7 LPZ127 439.3 499.4
53.5 3781.8 3083.3 3769.8 3848.4 1729.6 2902.2 LPZ128 1022.3 1063.8
447.9 3904.9 3753.5 5579.6 4534.2 2133.1 5843.6 LPZ131 249.6 373.8
27.3 2170.1 1551.6 1611.4 3077.3 2946 2409.5 LPZ133 325.3 328.7 42
2706.9 2358.1 2131.3 3257.2 2598.7 2144.3 LPZ136 263.6 384.1 0.9
2377.3 3386.4 2044.7 2726.4 2398.2 2570.6 LPZ137 351.6 402.6 91.9
3435.9 3122.8 2406.9 3054.7 3392.1 3216.6 LPZ138 1047.7 936 530.3
3920 3091.2 2507.9 4249.5 3109.9 4758.4 LPZ140 379.9 456.6 105.6
1936.7 3674 2908.4 3678.1 3663.2 4481.7 LPZ141 715.4 1341.8 536.4
4712.8 3615.6 3746 4815.5 4533 8229.3 LPZ143 3251.1 4810.8 2848.5
9675.1 8289.6 9486.6 12270.6 4424.9 28251.9 LPZ144 386.2 1338.3
294.1 3433.8 3015.7 3333.7 4618.9 3699.7 6825.1 LPZ145 277.5 1064
536.5 3072.3 1155.6 2977.7 3188.2 7388.2 3013.6 LPZ146 128.5 362.8
139.5 2224.4 1364 2125.6 2428.3 3465.8 2351.1 LPZ147 266.5 544.2
241.3 3318.1 2241.9 2162.6 2908.8 3252.6 2525.6 LPZ148 725.8 725.5
290.2 5002.2 3682.1 6542.3 6927.9 2903.1 10410.9 LPZ149 2492.7
3102.7 1052.7 8980.9 7669.2 7185.3 9921 7375.4 12848.8 LP2150
5257.3 7586.3 3221.3 15156.2 12776.8 7401.2 14686.5 1570 20635.9
LPZ151 1384 2049.2 1128.3 5576.7 3853.7 3780.3 7227.3 3206.7 7702.7
LPZ152 975.8 1300.2 546.4 5096.1 4344.9 4302.5 4960.3 1566.2 4223.4
LPZ153 13378.6 22235.1 10073.1 33495.1 50802.7 28291.5 29602.5
3224.1 40232.7 LPZ154 663.3 738 430.3 3847.3 3880.2 6253.9 4707.7
5341.9 3393.8 LPZ155 1121.7 612.7 748 3902.3 4289 4676 5328 6964.2
5353.5 LPZ157 1157 957.9 762.2 4218.8 4266.6 3777.2 4583.7 5017.5
4396.6 LPZ158 13278.5 17582.9 9898.7 32456.5 42805.1 24534.6
31442.6 4152.5 36015.3 LPZ162 3407.5 5943.1 5126.6 10324 11710.9
7727.6 7572.2 2312.6 9394.1 LPZ165 1419.4 1550 693 4519.9 3692.9
6102.1 5617.1 3061.3 4706.4 LPZ166 2642.1 3504.9 1288.6 7134.8
6994.5 5170.5 10453.3 4932.9 15993.7 LPZ167 980.4 1518.1 564.7
4902.6 4803.7 3343.6 4869.5 4421.2 6582.2 LPZ169 621.8 792.7 93.4
3562 4266.3 1794.9 3496.3 4238 3143.7 LPZ170 1009.3 1405.3 695.2
5793.7 4469.2 4952.7 6239.7 3914.1 6782.3 LPZ171 1064.6 968.6
1050.7 4110.8 3903.7 5467.2 5659 3164.1 6795.3 LPZ172 1233.3 1113.3
401.8 4207.9 7922 8417.5 10419.2 7983.5 15065.2 LPZ173 3333.8
5236.8 6072 9552 8880 8653.2 13461.6 2408.7 25719.1 LPZ174 486.7
1263.6 143.3 3318.2 2027 3632.3 4245.3 6086.4 8781.2 LPZ175 594.7
1487 520.8 3051.5 3610.8 1846.4 3642.9 4048.5 4329.5 LPZ177 167.3
481.7 234.5 1955.9 1139.2 907.8 1452.9 4462.5 1762 LPZ179 1231.4
1583.2 835.8 5029.7 3654 3871.7 3248.8 2741.9 2853.2 LPZ181 2259.5
4282.9 774.8 8911.6 8788.6 6833.3 5825.5 3398 3922.8 LPZ182 1194.7
2289.6 673.8 8220.3 5601.2 5869.3 5572 1808.4 7607.9 LPZ186 6255.6
6866.5 6841.1 24861.8 16742.4 23502.2 17304.6 1750.4 27328.9 LPZ189
27581.8 34787.4 31889 45673.7 106688 95043.9 46998.5 3548.1 67943.9
LPZ194 673.9 904.9 528.7 4064.5 3530.2 3160.6 5095.2 4531.1 3492.1
LPZ195 898.9 889.6 675.7 5606.3 4004.3 5230.5 5721.4 4908.1 4940.9
LPZ196 1073.7 1935.1 558.2 3941.1 3672.6 3681.6 5977.8 3059.7
5050.3 LPZ197 488.4 522.5 386.6 2613.4 1684.4 3541.4 3507.4 4980.8
2763.1 LPZ198 575.6 733.1 299.6 4152.3 2411 2916.8 3872.4 6530.7
2931.8 LPZ199 390.7 442.5 222.4 2704 2176.2 3159.7 2957.1 5357.6
2983.4 LPZ201 2255.7 3876.6 347.6 10222.9 6897.7 6294.3 7324.7 3549
3857 LPZ202 25939.2 34864.8 28937 43395.9 85136 71116.8 38688.5
2676.2 14449.1 LPZ203 4917.3 4458.1 2603.2 10348.3 6571.1 8325.7
11034 5453.1 6598 LPZ204 27637.2 31853.9 22475.7 43983.9 89520.6
46504.2 44333.5 7963.8 58054.1 LPZ205 1184 1084.2 327.7 3901.8 4402
3125 4598.2 4501.5 4714.2 LPZ206 1309.8 1509.5 367.3 3961.5 3983.3
3079.6 4196.2 3641.4 3247.6 LPZ207 27569.1 30446.2 27094.6 45211.6
90196.1 58153.8 46488.1 9879.3 64709.2 LPZ208 22722 28208.9 30019.5
40314 74354.1 37339 33919.6 3989.8 56683.6 LPZ210 1015.1 1789.8
196.8 5006.3 6159.7 3067.8 4944 4347.5 6846.4 LPZ211 277 327.3
519.4 2540.7 1788.5 3048.6 1110.8 2577.4 3506.6 LPZ212 1095.5 865.5
930.6 4051.6 4491.6 2857.6 6348.1 4078.7 16174.9 LPZ213 376.6 539.3
339.6 2503.1 1540.8 1333.7 3082.2 6024.3 4163.1 LPZ214 137.5 306.1
190.1 1915.6 866.1 1280.9 1240.8 6372.4 2111.8 LPZ215 3519.4 3120.9
3300.9 16939.5 15489.1 10948.6 12502 3515.1 16236.4 LPZ216 26761.3
34226.4 28477.9 42274.8 67630.3 41420.3 36331.4 1433.8 17109.4
LPZ217 15563.1 21739.4 12259 26824.9 34266.3 9429.4 28156.7 1339.2
41568.4 LPZ219 2404.9 3704.5 2084 8575.1 8573.2 6237 11757 3255.4
13484.9 LPZ220 3617.2 6998.6 7957.2 13960 9400.8 3432.3 10805.5
3551.7 12867.4 LPZ221 482.6 478 1405.6 3296.5 3079.8 3312.5 4143
4429.4 3267.3 LPZ222 318.3 524.5 406.6 3011.7 2309 3811.8 4199.4
5319.9 3292.3 LP2223 367.7 633.2 437.7 2752 1970.3 3767.4 2514.2
3672.3 2163.1 LP2224 337.9 1030.2 317.2 2701.2 1798.7 7393.6 2962.5
7876 3353.7 LPZ225 3288.1 3590.3 2912 8781.8 7400.7 2317.1 11370.2
3186.6 22244.6 LP2226 325.6 361.4 128 2467 1263.7 10190.1 1636.3
3808.8 1166.9 LP2227 2175.5 6375.8 458.8 6316.2 6632.1 9013.9
6614.3 3901.2 2588.2 LPZ228 2638 3701.7 500.1 5991 4819.8 5747.8
7102.5 3182.5 4185.4 LPZ231 1631.7 2090.4 260.7 5811.5 4749 2530.8
5033.1 3191.3 3810.5 LPZ233 1596.6 1223 296.6 4355.6 3818.9 2988.8
3749 3324.7 3855.2 LPZ234 1734.3 1479.2 219.6 5058.5 4614.4 2034.9
4992.1 1979.9 5152 LPZ235 626 635.9 185.9 4066.8 3255.5 4035.7
3368.7 2880.8 3643.5 LPZ237 1677.8 1385.3 847.4 4536 3702.8 2943.6
4886.5 2307.8 5136.8 LPZ239 673.4 407.8 245.8 2981.9 3199.2 2781.6
4235.6 2342.6 4863.7 LPZ240 387 247.4 254.8 2075.8 2317.4 2894
2721.7 2054.5 4317.9 LPZ241 258.3 337.8 110.9 3503.1 3829.6 22593.5
1889.5 1315.9 8842.6 LPZ242 4315.9 2560.2 22.5 12510.2 12605.3
2345.2 16197.1 1114.8 39684.4 LPZ243 174.8 274.4 23.1 2193.6 346.2
2395.7 1366.5 2568.2 3103.8 LPZ244 417.5 269.1 3458.5 3545.1 1831
2834.7 1781.1 7589.2 5662.7 LPZ246 889.5 918.7 2302.7 3920.3 3228.9
4409.3 3536 1258.8 2645.3 LPZ247 1203 2088.9 46.7 4956.9 4253.2
3559.7 4570.8 1702.6 3350.6 LPZ248 973.3 1338.1 86.2 3977.4 4392.8
2033 4094.9 2062.6 4279.9 LPZ249 361.3 324.3 206.7 1948.6 1764.3
2098.7 2762.1 1643 2862.4 LPZ250 267.6 487.8 118.7 2690.4 1522 2989
3121 1928.9 1809.7 LPZ251 245 279.7 168.5 1409.9 555 9932.3 2552.5
3050.3 1371.1 LPZ255 2021.3 2488.5 334.1 7289.5 7773.7 1269.1
9020.3 4492.4 10134.6 LPZ256 67.1 72.2 296.7 412.2 229.8 922.7
570.3 5040.6 1263.9 LPZ257 167.3 146.9 482.6 521.8 102.5 2699.4
599.8 2362.1 1553.4 LPZ258 247.5 236.5 69.7 1429.8 974.6 971.9
2668.7 2990.8 3445.9 LPZ260 98.1 188.8 463.1 377.1 337.2 880 808.9
1552 1084.6 LPZ261 73.7 20.5 386.3 1143.6 50.3 4443.8 903.3 2309
1341.6 LPZ264 482.7 528.5 1151.3 3659.7 1972.6 9892.4 2831.4 1584.4
3208 LPZ265 534.6 647 457.6 4473 4089.9 656 5899.6 2972.2 5649.8
LPZ266 16.9 61.2 1062.3 876.1 1183.4 1624.3 663.3 622.4 1609.7
LPZ268 143.7 142.9 255 1983.5 810.3 9809.8 1293.5 1757.3 2177
LPZ269 1747.5 1271.8 1636.4 7364.1 5108.7 6903.3 11401.2 3774.1
14643.8 LPZ270 373.8 77.9 1901.2 5015 3872.6 3485.9 5621.1 4284.8
5197.8 LPZ271 705 473.1 315.6 2863.8 2625.4 2120.5 4048.2 1424.9
4291.1 LPZ272 2809.4 2423.8 300 5056.2 2463 3534.6 3496.9 609.7
3996.8 LPZ273 219.8 162.4 242.4 90.2 130.2 3251.3 166 1193.5 1836.5
LPZ274 489.2 367.7 284.6 991 1104.2 395.9 2282.3 747.2 4535.7
LPZ275 93.5 140 156.8 433.3 217.1 0 837.9 1056.2 2352.3 LPZ276 53
109.7 106.8 0 0 0 369.1 1303.6 1897.6 LPZ277 105.9 159.4 68.7 0 0
230.1 236.2 706.1 1337.9 LPZ278 65.7 48.3 156.4 0 0 1788.2 406.3
1442.3 1564.1 LPZ279 214.7 212.2 75.3 1356.3 790.8 5213.3 1722.4
496 2426.2 LPZ280 156.2 247.7 1553.6 3510.5 2515.1 289.7 3182.1
612.7 3123.7 LPZ281 34.6 92.1 73 0 0 1648.3 565.5 522.8 543.3
LPZ282 200.9 187.7 218.1 536.6 205.9 7324.8 1145.4 2977.5 957.5
LPZ283 1833.5 1880 775.8 3303.2 5113.2 527.6 5281.2 324.5 3806.8
LPZ284 215 0.6 148.8 0 29.6 848 408.4 148.8 1294.3 LPZ286 219.7
21.9 13.6 234.9 78.6 2703.6 198.4 947.9 1233.2 LPZ287 112.6 126.2
36.5 1170.3 459.9 306.3 157.5 1173.7 1821.3 LPZ288 23.6 62.2 37.5
774.1 639.3 792.6 715.8 1422.6 1169 LPZ289 44.1 13.1 107.4 323.4 95
9975.4 889.5 2240.6 1894.9 LPZ290 1324.7 1572.4 1838 6941.6 4616.9
2995.3 11538.8 407.1 12699.5 LP2293 45.2 246.3 145.6 2785.5 1923.1
0 3185.6 0 3550.4 LPZ294 0 19.8 0 403.3 280.4 89.1 785.9 551.6
1378.4 LPZ295 40 24.5 0 169.9 26 1324.9 1058 848.8 1406.5 LPZ297
385.6 127.6 17.4 1238.5 941.5 0 2680.9 2084.3 4065.3 LPZ299 106.9
36.2 0 0 926.2 0 1060.7 1854.9 1575.9 LPZ300 73.2 93.2 80.2 0
1143.6 1053.3 1034.5 2304.9 2120.8 LPZ301 126.2 0 5.8 161.2 1245.7
516.3 1612 761.3 2826.1 LPZ303 83.1 488.8 98.6 0 73.5 979.9 538.7
510.7 1214.7 LPZ304 213.7 498.3 137.6 1028.6 0 5405.8 860 2212.1
2201 LPZ306 1439.4 1735.3 2526.4 4212.7 3140.4 2090.1 8128.5 4874.6
14413.9 LPZ307 534.1 710.5 515.5 2785.3 734 0 2137.3 1692.8 3540.3
LPZ308 116 304.4 137.7 151.8 28.2 364.2 621.1 631.4 851.2 LPZ309
80.1 137.2 92.7 0 0 2648.1 529.4 192.6 735 LPZ310 430.8 584.9 799.2
1887.2 1887.1 6161.2 2974.3 3575 2426.6 LPZ311 690.5 995.7 208.4
3725.8 2843.8 0 4329.3 3620.8 4170.1 LPZ312 109.8 334.2 34 72.5 4.5
1489.3 140.1 431.6 744.8 LPZ314 26.5 200.1 3.3 181.2 0 1231.5 331.5
440.1 804.6 LPZ315 305.8 211.3 147.5 811.2 1008.1 3797 2231.8
1438.8 1881.8 LPZ318 621.3 715 337 3488.2 2480.9 781.9 4326.1
4824.7 6969.2 LPZ320 214.8 92.2 9.9 1170.9 54.5 4501.5 1122.3
1169.4 1696.6 LPZ321 880.4 755.2 1899.3 6166.2 5105.8 411.6 6096.5
4853.6 6057.2
[0239]
3TABLE III LSC Media Multiplication Media Maturation Media
Components (mg/L) 16 1133 923 NH.sub.4NO.sub.3 603.8 603.8 200.0
KNO.sub.3 909.9 909.9 454.95 KH.sub.2PO.sub.4 136.1 136.1 136.1
Ca(NO.sub.3).sub.2.4H.sub.2- O 236.2 236.2 59.05
MgSO.sub.4.7H.sub.2O 246.5 246.5 246.5 Mg(NO.sub.3).sub.2.6H.sub.2O
256.5 256.5 256.5 MgCl.sub.2.6 H.sub.2O 101.7 101.7 101.7 Kl 4.15
4.15 4.15 H.sub.3BO.sub.3 15.5 15.5 7.75 MnSO.sub.4.H.sub.2O 10.5
10.5 10.5 ZnSO.sub.4.7 H.sub.2O 14.4 14.4 14.4 NaMoO.sub.4.2
H.sub.2O 0.125 0.125 0.125 CuSO.sub.4.5 H.sub.2O 0.125 0.125 0.125
CoCl.sub.2.6 H.sub.2O 0.125 0.125 0.125 FeSo.sub.4.7 H.sub.2O 6.95
6.95 41.7 Na.sub.2EDTA 9.33 9.33 55.9 Sucrose 30,000 30,000 --
Maltose -- -- 20,000 myo-Inositol 1,000 1,000 100 Casamino acids
500 500 500 L-Glutamine 450 450 450 Thiamine.HCl 1.0 1.0 1.0
Pyridoxine.HCl 0.5 0.5 0.5 Nicotinic acid 0.5 0.5 0.5 Glycine 2.0
2.0 2.0 2,4-D 1.1 1.1 -- BAP 0.45 0.45 -- Kinetin 0.43 0.43 --
Polyethylene glycol -- -- 130,000 ABA -- 5.2 5.2 Gelrite 2,500*
2,500* 2,500 pH 5.7 5.7 5.7 *For solid media only
[0240]
4TABLE IV Description of clones used in hybridization study shown
in FIG. 9. ID with Clone # Homology Description Arabidopsis Score
E-value PC04B12 Lotan et al.. 1998. Arabidopsis Required for embryo
79% ID, 171 7e-44 (`LEC` in LEAFY COTYLEDON 1 is maturation &
Cotyledon 93% + ve figure) sufficient to Induce Embryo identity.
Ectopic over 96aa Development in Vegetative expression induces
Cells. Cell 93: 1195-1205 embryonic differentiation traits in
transgenic seedlings. ST17B05 PICLKE/CDH3, Chromatin The pickle
mutants 50% ID, 166 1e-41 (`PLK` in remodelling. Ogas et al. 1999.
express embryonic traits 74% + ve figure) PICKLE is a CHD3
chromatin- after germination. over 155aa remodeling factor that
Represses lec regulates the transition from expression embryonic to
vegetative development in Arabidopsis. PNAS. 96(24): 13839-13844
PC08C06 FIE, fertilization-independent Fie mutants initiate 61% ID
92 8e-20 (`FIE` in endosperm protein. Ohad, et al endosperm
development 75% + ve figure.) 1999. Mutations in FIE, a WD w/o
fertilization over 67aa polycomb group gene, allow endosperm
development without fertilization. Plant Cell 11 (3), 407-416
[0241]
5TABLE V 488 499 499 500 500 (Liquid (Liquid (Liquid (Liquid
(Liquid) Cell Line Suspen- Suspen- Suspen- Suspen- Suspen- (Stage
of sion sion sion sion sion Develop- Culture: Culture: Culture:
Culture: Culture: 260 260 ment) Stage 1-3) Stage 1-3) Stage 1-3)
Stage 1-3) Stage 1-3 (Stage 7) (Stage 9) Media 1133 16 1133 16 1133
Maturation maturation # Embyros 1 118.5 2 187.75 3 Na na `FIE` ++++
+ +++ +++ +++ +++ +++ `LEC` 4 ++ 5 ++ 6 + + `PKL` ++++ + +++ +++
+++ +++ Table 5. Table of data from Fig. 9a & b. Numbers (488,
499, 500, 260) refer to different cell lines Liquid Suspension
Culture contains early-stage embryos (stage 1-3) Embryo number
refers to the number of late-stage (stage 8-9) embryos # produced
by each cell line when matured according to Pullman and Webb
(1994). + = low expression, ++ medium level of mRNA, +++ = high
level of mRNA, ++++ = very high level of mRNA. Circles # around
certain + signs, see text. Na = not applicable. Levels of mRNA are
relative and refer to the experiment depicted in Fig. 9a &
b.
[0242]
Sequence CWU 0
0
* * * * *
References