U.S. patent application number 11/832019 was filed with the patent office on 2008-03-06 for polynucleotides and polypeptides encoding receptors.
This patent application is currently assigned to Human Genome Sciences, Inc.. Invention is credited to Reiner Gentz, Jian Ni, Craig A. Rosen.
Application Number | 20080058247 11/832019 |
Document ID | / |
Family ID | 26710688 |
Filed Date | 2008-03-06 |
United States Patent
Application |
20080058247 |
Kind Code |
A1 |
Ni; Jian ; et al. |
March 6, 2008 |
Polynucleotides and Polypeptides Encoding Receptors
Abstract
Receptor polypeptides and polynucleotides and methods for
producing such polypeptides by recombinant techniques are
disclosed. Also disclosed are methods for utilizing receptor
polypeptides and polynucleotides in the design of protocols for the
treatment of diseases and diagnostic assays for such
conditions.
Inventors: |
Ni; Jian; (Germantown,
MD) ; Rosen; Craig A.; (Laytonsville, MD) ;
Gentz; Reiner; (Belo Horizonte-Mg, BR) |
Correspondence
Address: |
HUMAN GENOME SCIENCES INC.;INTELLECTUAL PROPERTY DEPT.
14200 SHADY GROVE ROAD
ROCKVILLE
MD
20850
US
|
Assignee: |
Human Genome Sciences, Inc.
Rockville
MD
20850
|
Family ID: |
26710688 |
Appl. No.: |
11/832019 |
Filed: |
August 1, 2007 |
Related U.S. Patent Documents
|
|
|
|
|
|
Application
Number |
Filing Date |
Patent Number |
|
|
11041419 |
Jan 25, 2005 |
|
|
|
11832019 |
Aug 1, 2007 |
|
|
|
10156136 |
May 29, 2002 |
|
|
|
11041419 |
Jan 25, 2005 |
|
|
|
09764452 |
Jan 19, 2001 |
|
|
|
10156136 |
May 29, 2002 |
|
|
|
09010146 |
Jan 21, 1998 |
|
|
|
09764452 |
Jan 19, 2001 |
|
|
|
60034204 |
Jan 21, 1997 |
|
|
|
60034205 |
Jan 21, 1997 |
|
|
|
Current U.S.
Class: |
514/44R ;
435/346; 435/6.14; 435/69.1; 435/7.21; 530/350; 530/387.9;
536/23.1; 536/23.5 |
Current CPC
Class: |
A61P 35/00 20180101;
C12N 9/6489 20130101; C07K 14/70535 20130101; A61K 38/00 20130101;
C12N 2799/026 20130101; C12N 9/6475 20130101; C12N 9/6421 20130101;
C12N 9/6491 20130101; C07K 14/70503 20130101; C07K 14/705 20130101;
C07K 14/47 20130101; A61P 43/00 20180101 |
Class at
Publication: |
514/002 ;
435/346; 435/006; 435/069.1; 435/007.21; 514/044; 530/350;
530/387.9; 536/023.1; 536/023.5 |
International
Class: |
A61K 31/70 20060101
A61K031/70; A61K 38/00 20060101 A61K038/00; A61P 43/00 20060101
A61P043/00; C07H 21/04 20060101 C07H021/04; C07K 14/00 20060101
C07K014/00; C07K 16/18 20060101 C07K016/18; C12N 5/00 20060101
C12N005/00; G01N 33/00 20060101 G01N033/00 |
Claims
1. An isolated polynucleotide comprising a nucleotide sequence that
has at least 80% identity over its entire length to a nucleotide
sequence selected from the group consisting of: (a) a nucleotide
sequence encoding the receptor polypeptide of SEQ ID NO:Y; (b) a
nucleotide sequence having at least 80% identity to a nucleotide
sequence encoding the receptor polypeptide expressed by the cDNA
insert deposited at the ATCC; or (c) a nucleotide sequence
complementary to said isolated polynucleotide.
2. The polynucleotide of claim 1 wherein said polynucleotide
comprises the nucleotide sequence contained in SEQ ID NO:X encoding
the receptor polypeptide of SEQ ID NO:Y.
3. The polynucleotide of claim 1 wherein said polynucleotide
comprises a nucleotide sequence that is at least 80% identical to
that of SEQ ID NO:X over its entire length.
4. The polynucleotide of claim 3 which is polynucleotide of SEQ ID
NO: X.
5. The polynucleotide of claim 1 which is DNA or RNA.
6. A DNA or RNA molecule comprising an expression system, wherein
said expression system is capable of producing a receptor
polypeptide comprising an amino acid sequence, which has at least
80% identity with the polypeptide of SEQ ID NO:Y when said
expression system is present in a compatible host cell.
7. A host cell comprising the expression system of claim 6.
8. A process for producing a receptor polypeptide comprising
culturing a host of claim 7 under conditions sufficient for the
production of said polypeptide and recovering the polypeptide from
the culture.
9. A recombinant host cell produced by a method comprising
transforming or transfecting a host cell with the expression system
of claim 6 such that the host cell, under appropriate culture
conditions, produces a receptor polypeptide.
10. A receptor polypeptide comprising an amino acid sequence which
is at least 80% identical to the amino acid sequence of SEQ ID NO:Y
over its entire length.
11. The polypeptide of claim 10 which comprises the amino acid
sequence of SEQ ID NO:Y.
12. An antibody immunospecific for the receptor polypeptide of
claim 10.
13. A method for the treatment of a subject in need of enhanced
activity or expression of receptor polypeptide of claim 10
comprising: (a) administering to the subject a therapeutically
effective amount of an agonist to said receptor; and/or (b)
providing to the subject polynucleotide of claim 1 in a form so as
to effect production of said receptor activity in vivo.
14. A method for the treatment of a subject having need to inhibit
activity or expression of the receptor polypeptide of claim 10
comprising: (a) administering to the subject a therapeutically
effective amount of an antagonist to said receptor; and/or (b)
administering to the subject a nucleic acid molecule that inhibits
the expression of the nucleotide sequence encoding said receptor;
and/or (c) administering to the subject a therapeutically effective
amount of a polypeptide that competes with said receptor for its
ligand.
15. A process for diagnosing a disease or a susceptibility to a
disease in a subject related to expression or activity of the
receptor polypeptide of claim 10 in a subject comprising: (a)
determining the presence or absence of a mutation in the nucleotide
sequence encoding said receptor polypeptide in the genome of said
subject; and/or (b) analyzing for the presence or amount of the
receptor polypeptide expression in a sample derived from said
subject.
16. A method for identifying agonists to the receptor polypeptide
of claim 10 comprising: (a) contacting cells produced by claim 9
with a candidate compound; and (b) determining whether the
candidate compound effects a signal generated by activation of the
receptor polypeptide.
17. An agonist identified by the method of claim 16.
18. The method for identifying antagonists to the receptor
polypeptide of claim 10 comprising: (a) contacting said cell
produced by claim 9 with an agonist; and (b) determining whether
the signal generated by said agonist is diminished in the presence
of a candidate compound.
19. An antagonist identified by the method of claim 18.
Description
CROSS-REFERENCE TO RELATED APPLICATIONS
[0001] This application is a continuation of application Ser. No.
11/041,419, filed Jan. 25, 2005, which is a continuation of
application Ser. No. 10/156,136, filed May 29, 2002, which is a
continuation of application Ser. No. 09/764,452, filed Jan. 19,
2001, which is a continuation of application Ser. No. 09/010,146,
filed Jan. 21, 1998, which claims the benefit under 35 U.S.C.
.sctn. 119(e) of provisional Application No. 60/034,204, filed Jan.
21, 1997 and provisional Application No. 60/034,205, filed Jan. 21,
1997; each of which is hereby incorporated by reference in its
entirety.
STATEMENT UNDER 37 C.F.R. .sctn. 1.77(b)(4)
[0002] This application refers to a "Sequence Listing" listed
below, which is provided as a text document. The document is
entitled "PF354C4_SeqListing.txt" (112,019 bytes, created Jul. 31,
2007), and is hereby incorporated by reference in its entirety
herein.
BACKGROUND OF THE INVENTION
[0003] Receptor proteins are found on the membrane of the cells and
are generally involved in signal transduction. There are many types
of receptor proteins, and for convenience, these proteins are
grouped in families based on similarity in structure and
function.
[0004] For example, the TM4SF superfamily of cell surface proteins,
also known as the tetraspan receptor superfamily, is comprised of
at least seventeen individual gene products (these include CD9,
CD20, CD37, CD53, CD63, CD81, CD82, A15, CO-029, Sm23, RDS, Uro B,
Uro A, SAS, Rom-1, PETA3, and YKK8). The TM4SF superfamily is the
second largest group in the CD antigen superfamily. Each member of
the TM4SF superfamily can be characterized by several putative
physical features including four highly conserved transmembrane
domains, two divergent extracellular loops, and two short and
highly divergent cytoplasmic tails. Expression patterns for members
of the TM4SF superfamily tend to be rather broad and can vary
widely between members. The functional roles of TM4SF superfamily
members are primarily associated with signal transduction events
and pathways, but also include cell adhesion in platelets and other
lymphocytic and non-lymphocytic cell lines, as well as cell
motility, proliferation, and metastasis. In addition, recent
evidence suggests that a subset of the members of the TM4SF
superfamily may function as potassium channel molecules.
[0005] One member of the TM4SF family, CD20, is a four membrane
spanning domain cell surface phosphoprotein expressed exclusively
on B lymphocytes. Although the precise functional role of CD20 has
yet to be determined, it is thought to function primarily as a
receptor during B-cell activation. Furthermore, a large number of
experimental observations suggest several additional speculative
roles for the CD20 molecule. For example, CD20-specific
immunoprecipitation of biochemically cross-linked plasma membrane
proteins suggests that CD20 assumes a multimeric structural
conformation characteristic of other previously described membrane
channel proteins. Further experimentation has revealed that
expression of exogenous CD20 on the cell surface specifically
increases Ca.sup.2+ conductance across the plasma membrane.
Together, these results suggest that CD20 complexes may function as
B-cell specific Ca.sup.2+ ion channels. In addition, monoclonal
antibodies raised against CD20 have been used to stimulate resting
B-cells to transition out of the G0/G1 segment of the cell cycle.
It has also been demonstrated that CD20 is associated with both
serine and tyrosine kinases and, more specifically, that CD20 is
associated, although not directly, with the Src family of tyrosine
kinases including p56/531yn, p561ck, and p59fyn.
[0006] A second example of a receptor subfamily, called
sialoadhesin molecules, belongs to the Ig superfamily of
receptor-like molecules. The more than 100 members of the Ig
superfamily are generally considered to engage in specific
cell-cell interactions through which intercellular communication
may occur. In addition to classical protein-protein interactions,
intercellular communication may also be mediated through
protein-carbohydrate interactions. In fact, all members of the
sialoadhesin family of the Ig superfamily are capable of mediating
protein-sialic acid binding interactions. To date, only a small
number of proteins have been assigned to the sialoadhesin family
including sialoadhesin, CD33, CD22, the myelin-associated
glycoprotein (MAG), and the Schwann cell myelin protein (SMP). Each
of these proteins is expressed in a restricted subset of cell
types. For example, CD22 and CD33 are expressed exclusively by
B-lymphocytes and cells of the myelomonocytic lineage,
respectively.
[0007] Similarly, galectins are a family of the lectin superfamily
of carbohydrate-binding proteins which have a high affinity for
b-galactoside sugars. Although a large number of glycoproteins
containing b-galactoside sugars are produced by the cell, only a
few will bind to known galectins in vitro. Such apparent binding
specificity suggests a highly specific functional role for the
galectins. Galectin 1 (conventionally termed LGALS1 for lectin,
galactoside-binding, soluble A) is thought to specifically bind
laminin, a highly polylactosaminated cellular glycoprotein, as well
as the highly polylactosaminated lysosome-associated membrane
proteins (LAMPs). Galectin 1 has also been shown to bind
specifically to a lactosamine-containing glycolipid found on
olfactory neurons and to integrin a.sub.7b.sub.1 on skeletal muscle
cells. Galectin 3 has also been observed to bind specifically to
laminin, immunoglobulin E and its receptor, and bacterial
lipopolysaccharides.
[0008] Various galectins have been shown to function in the
mechanisms of intercellular communication. For example, depending
on cell type, galectin 1 has been observed to modulate cell
adhesion either positively or negatively. More specifically,
galectin 1 appears to inhibit cell adhesion of skeletal muscle
presumably by galectin 1-mediated disruption of laminin-integrin
a.sub.7b.sub.1 interactions. Alternatively, galectin 1 appears to
promote cell adhesion in several non-skeletal muscle cell types
examined presumably by a glycoconjugate cross-linking mechanism.
Galectin 3 has also been observed to function in modulating
cell-adhesion, as well as in the activation of certain immune cells
by cross-linking IgE and IgE receptors. In addition, galectins have
been observed to be involved in the regulation of immune cell
activity, as well as in such diverse processes as cell adhesion,
proliferation, inflammation, autoimmunity, and metastasis of tumor
cells. Furthermore, a galectin-like antigen designated HOM-HD-21
was recently found to be highly expressed in a Hodgkin's Disease
cDNA library. Very recently, a novel galectin, termed PCTA-1, was
identified as a specific cell surface marker on human prostate
cancer cell lines and patient-derived carcinomas. Galectins have
also been found to function intracellularly as a component of
ribonucleoprotein complexes. Finally, galectins 1 and 3 have each
been found to modulate T-cell growth and apoptosis by interaction
with CD45 and possibly Bcl2, respectively.
[0009] A relatively new family of cell-surface proteins has been
identified and termed the Ly6 superfamily. The members of this
family include murine and human SCA-2, rat Ly-6 (also termed ThB),
human CD59 [also known as protectin or membrane attack complex
inhibition factor (MACIF)], and E48 antigen. The determination of
an initial functional role for SCA-2 may lie in an analysis of its
expression profile with regard to the complex process of
hematopoiesis. SCA-2 is highly expressed in early thymic precusor
cells. In turn, progeny of the intrathymic precusor population
continue to express SCA-2, but only until the point of transition
occurs from blast cell to small cell. Further experimental evidence
demonstrates that mature thymocytes and peripheral T-cells do not
express detectable levels of SCA-2, whereas mature, peripheral
B-cells do continue to express SCA-2. As a result, it seems very
likely that SCA-2 plays an important role in thymocyte maturation
and differentiation. A plausible explanation for this functional
hypothesis is that SCA-2 may act as a receptor for a unknown
cytokine which regulates thymocyte maturation and
differentiation.
[0010] In addition, CD59 is a recently identified integral membrane
protein which appears to be involved in the regulation of
complement. Recent studies show that the CD59 antigen may prevent
damage from complement C5b-9 and protect astrocytes during
inflammatory and infectious disorders of the nervous system.
Expression of recombinant human CD59 on porcine donor organs have
been shown to prevent complement-mediated lysis and activation of
endothelial cells that leads to hyperacute rejection. Recently,
researchers at Alexion Pharmaceuticals (New Haven, Conn.) reported
on the production of transgenic pigs which expressed human CD59. In
these animals, xenogeneic organs were resistant to hyperacute
rejection. (Fodor, et al., "Expression of a functional human
complement inhibitor in a transgenic pig as a model for the
prevention of xenogeneic hyperacute organ rejection," Proc. Natl.
Acad. Sci., 91:1153-11157 (1994).) The same company also reported
that expression of recombinant transmembrane CD59 in paroxysmal
nocturnal hemoglobinuria (PNH) B-cells confers resistance to human
complement. (Rother et al., "Expression of recombinant
transmembrane CD59 in paroxysmal nocturnal hemoglobinuria B-cells
confers resistance to human complement," Blood, 84:2604-2611
(1994).) PNH is an acquired hematopoietic disorder characterized by
complement-mediated hemolytic anemia, pancytopenia, and venous
thrombosis. It is thought that retroviral gene therapy with this
molecule could provide a treatment for PNH patients.
[0011] A final Ly6 superfamily member, the E48 antigen, is involved
in intercellular adhesion between keratinocyte cells of the
squamous epithelium. Such keratinocytes are attached to adjoining
cells by large numbers of desmosomes, which are thought to play a
role in the transition of transformed keratinocytes to metastatic
tumor cells. Treatment with a monoclonal antibody raised against
the E48 antigen has been successful in the eradication of residual,
postoperative squamous cell carcinoma cells of the upper
aerodigestive tract in several in vivo models and, to some degree,
in humans. (van Dongen, et al., "Progress in radioimmunotherapy of
head and neck cancer," Oncol. Rep. 1:259-264 (1994).) The gene
encoding the E48 antigen has been mapped to the q24-qter region of
human chromosome 8. Interestingly, a number of human diseases have
been mapped to this region of chromosome 8 including Langer-Giedion
syndrome, brachio-otorhinolaryngeal syndrome, trichorhinolaryngeal
syndrome, and epidermolysis bullosa simplex.
[0012] A further example of a receptor family includes the
prohibitin receptors. The prohibitin gene product is expressed in a
wide variety of tissues and has been implicated as a component of a
number of anti-proliferative mechanisms. The prohibitin gene
encodes a 30 kD postsynthetically modified polypeptide located
primarily in the mitochondria, but also may be associated with the
IgM receptor on the B-cell plasma membrane. The protein
functionally inhibits DNA synthesis and entry into S phase of the
cell cycle by an unknown mechanism. Interestingly, although the
prohibitin gene product is hypothesized to be involved in the
maintenance of senescence and the prevention of cancer, one study
found that, although somatic mutations in the prohibitin gene were
present in a small number of breast cancers, no mutations were
identified in any other breast, ovary, liver, and lung cancers
examined. (Sato et al., Genomics 17:762-764 (1993).) However, the
prohibitin gene has been mapped to human chromosome 17q12-21, the
same region thought to contain the gene involved in sporadic breast
cancer. Furthermore, DNA sequence analysis of the prohibitin gene
identified somatic mutation in 4 of 23 cases of sporadic breast
cancer examined. Thus, prohibitin family members may be involved in
the development of cancer.
[0013] Moreover, the EGFR family of plasma membrane proteins are an
integral component of normal cellular proliferation and in the
pathogenesis of the cancerous state. The family is relatively small
and includes the EGFR, .alpha.-erbB-2, c-erbB-3, and others.
Various cancers are correlated with aberrant expression of one or
more of these genes. A number of ligands have been identified which
bind to the EGFR-like receptors listed above including TGF-a,
heparin-binding EGF, amphiregulin, criptoregulin, heregulin, and
others. A large fraction of adenocarcinomas examined to date,
especially those of the breast, colon, and pancreas, are typified
by the amplification or overexpression of the c-erbB-2 gene. EGF,
or an analogous ligand, initiates the cellular growth factor
response by binding to the EGFR, or EGFR-related, receptor.
Following the binding event, the receptor molecule dimerizes
activating its intracellular tyrosine kinase domain. This event
results in the phosphorylation of specific tyrosine residues near
the carboxy terminus of the receptor. The diversity of signals able
to be transduced through the relatively small number of
EGFR-related receptor molecules is amplified considerably by the
recent finding that EGFR-like receptor molecules can function when
dimerized with other EGFR family members forming heterodimers.
[0014] Members of the EGFR-related family of integral membrane
proteins have been implicated in the pathogenesis of a number of
human disease-states. For example, a mutation in the EGFR itself
appears to play an important role in the development of
glioblastomas. (Sang et al., J. Neurosurg 82:841-846 (1995).) The
EGFR gene is amplified or overexpressed in the majority of primary
human glioblastomas. Although not conferring a distinct advantage
on cell growth, an increase in EGFR expression was found to confer
an increase in the ability of glioma cells to maintain
anchorage-independent growth in soft agar especially in response to
EGF and retinoic acid. Anchorage-independent growth in vitro
correlates highly with tumorigenicity in vivo, therefore, it is
likely that cells which express abnormally high levels of EGFR in
human glioblastoma cells may be involved in the high potential for
these cells to cause tumors in vivo.
[0015] Moreover, overexpression or amplification of c-erbB-2 has
been reported to be involved in a high number adenocarcinomas,
particularly of the breast, colon, and pancreas, and in a small
proportion of ovarian carcinomas.
[0016] Thus, there is a clear need for identifying and exploiting
novel members of the receptor families, such as those described
above. Although structurally related, these receptors will likely
possess diverse and multifaceted functions in a variety of cell and
tissue types. Receptor type molecules should prove useful in target
based screens for small molecules and other such pharmacologically
valuable factors. Monoclonal antibodies raised against such
receptors may prove useful as therapeutics in an anti-tumor,
diagnostic, or other capacity. Furthermore, receptors described
here may prove useful in an active or passive immunotherapeutical
role in patients with cancer or other immunocompromised disease
states.
BRIEF SUMMARY OF THE INVENTION
[0017] This invention relates to newly identified polynucleotides
and the polypeptides encoded by them, the use of such
polynucleotides and polypeptides, and their production. More
particularly, the polynucleotides and polypeptides of the present
invention relate to specific receptor families described in the
specification and known in the art. The invention also relates to
inhibiting or activating the action of such polynucleotides and
polypeptides.
[0018] In one aspect, the invention relates to receptor
polypeptides and polynucleotides, as well as the methods for their
production. Another aspect of the invention relates to methods for
using such receptor polypeptides and polynucleotides. Such uses
include the treatment of the specified diseases, among others. In
still another aspect, the invention relates to methods to identify
agonists and antagonists using the materials provided by the
invention, and treating conditions associated with receptor
imbalance with the identified compounds. Yet another aspect of the
invention relates to diagnostic assays for detecting diseases
associated with inappropriate receptor activity or levels.
BRIEF DESCRIPTION OF THE DRAWINGS
[0019] FIG. 1A-1B shows an amino acid sequence alignment of Clone
ID HMACR70 (SEQ ID NO:18) versus OB-1 (SEQ ID NO:33) (shaded boxes
indicate identical amino acid residues, non-shaded boxes indicate
conservative substitutions).
[0020] FIG. 2 shows an amino acid sequence alignment of Clone ID
HTEDK48 (SEQ ID NO:19) versus MRC-OX44 (SEQ ID NO:34) and PETA-3
(SEQ ID NO:35) (shaded boxes indicate identical amino acid
residues, non-shaded boxes indicate conservative
substitutions).
[0021] FIG. 3 shows an amino acid sequence alignment of Clone ID
HPWAE25 (SEQ ID NO:20) versus NAG-2 (SEQ ID NO:36) and TALLA-1 (SEQ
ID NO:37) (shaded boxes indicate identical amino acid residues,
non-shaded boxes indicate conservative substitutions).
[0022] FIG. 4 shows an amino acid sequence alignment of Clone ID
HTPEF86 (SEQ ID NO:21) versus B1 (SEQ ID NO:38) (shaded boxes
indicate identical amino acid residues, non-shaded boxes indicate
conservative substitutions).
[0023] FIG. 5 shows an amino acid sequence alignment of Clone ID
HSBBF02 (SEQ ID NO:22) versus TALLA-1 (SEQ ID NO:37) (shaded boxes
indicate identical amino acid residues, non-shaded boxes indicate
conservative substitutions).
[0024] FIG. 6 shows an amino acid sequence alignment of Clone ID
HLTAH80 (SEQ ID NO:23) versus TALLA-1 (SEQ ID NO:37) (shaded boxes
indicate identical amino acid residues, non-shaded boxes indicate
conservative substitutions).
[0025] FIG. 7 shows an amino acid sequence alignment of Clone ID
HTPBA27 (SEQ ID NO:24) versus NAG-2 (SEQ ID NO:36) (shaded boxes
indicate identical amino acid residues, non-shaded boxes indicate
conservative substitutions).
[0026] FIG. 8 shows an amino acid sequence alignment of Clone ID
HAIDQ59 (SEQ ID NO:25) versus CD9 (SEQ ID NO:39) (shaded boxes
indicate identical amino acid residues, non-shaded boxes indicate
conservative substitutions).
[0027] FIG. 9 shows an amino acid sequence alignment of Clone ID
HHFEK40 (SEQ ID NO:26) versus PETA-3 (SEQ ID NO:35) (shaded boxes
indicate identical amino acid residues, non-shaded boxes indicate
conservative substitutions).
[0028] FIG. 10 shows an amino acid sequence alignment of Clone ID
HGBGV89 (SEQ ID NO:27) versus L6H (SEQ ID NO:40) (shaded boxes
indicate identical amino acid residues, non-shaded boxes indicate
conservative substitutions).
[0029] FIG. 11 shows an amino acid sequence alignment of Clone ID
HUVBB80 (SEQ ID NO:28) versus L6 (SEQ ID NO:41) (shaded boxes
indicate identical amino acid residues, non-shaded boxes indicate
conservative substitutions).
[0030] FIG. 12 shows an amino acid sequence alignment of Clone ID
HJACE54 (SEQ ID NO:29) versus rGALECTIN-5 (SEQ ID NO:42) and
hGALECTIN-8 (SEQ ID NO:43) (shaded boxes indicate identical amino
acid residues, non-shaded boxes indicate conservative
substitutions).
[0031] FIG. 13 shows an amino acid sequence alignment of Clone ID
HROAD63 (SEQ ID NO:30) versus E48 (SEQ ID NO:44) (shaded boxes
indicate identical amino acid residues, non-shaded boxes indicate
conservative substitutions).
[0032] FIG. 14 shows an amino acid sequence alignment of Clone ID
HMWGS46 (SEQ ID NO:31) versus B-cell Receptor Associated Protein
(shaded boxes indicate identical amino acid residues, non-shaded
boxes indicate conservative substitutions).
[0033] FIG. 15 shows an amino acid sequence alignment of Clone ID
HNFGW06 (SEQ ID NO:32) versus EGFR (SEQ ID NO:46) (shaded boxes
indicate identical amino acid residues, non-shaded boxes indicate
conservative substitutions).
DETAILED DESCRIPTION OF THE INVENTION
Definitions
[0034] The following definitions are provided to facilitate
understanding of certain terms used frequently herein.
[0035] "Receptor" refers, among others, to a polypeptide comprising
the amino acid sequence set forth in SEQ ID NO:Y, or an allelic
variant thereof.
[0036] "Receptor Activity" or "Biological Activity of the Receptor"
refers to the metabolic or physiologic function of said receptor
including similar activities or improved activities or these
activities with decreased undesirable side-effects. Also included
are antigenic and immunogenic activities of said receptor.
[0037] "Receptor gene" refers to a polynucleotide comprising the
nucleotide sequence set forth in SEQ ID NO:X or allelic variants
thereof and/or their complements.
[0038] "SEQ ID NO:X" comprises all or a substantial portion of the
polynucleotide encoding each receptor of the invention. The value X
for the nucleotide sequence is an integer specified in Table 1.
This nucleotide sequence was translated into the receptor
polypeptide identified in Table 1 as "SEQ ID NO:Y," where the value
of Y for each receptor polypeptide is an integer defined in Table
1.
[0039] The invention further provides a composition of matter
comprising a nucleic acid molecule which comprises a human cDNA
clone identified by a cDNA Clone ID (Identifier) in Table 1, which
DNA molecule is contained in the material deposited with the
American Type Culture Collection ("ATCC") and given the ATCC
Deposit Number shown in Table 1 for that cDNA clone. The ATCC is
located at American Type Culture Collection (ATCC), 12301 Park Lawn
Drive, Rockville, Md. 20852, USA. The deposit has been made under
the terms of the Budapest Treaty on the international recognition
of the deposit of micro-organisms for purposes of patent procedure.
The strain will be irrevocably and without restriction or condition
released to the public upon the issuance of a patent. The deposit
is provided merely as convenience to those of skill in the art and
is not an admission that a deposit is required for enablement, such
as that required under 35 U.S.C. .sctn. 112. The nucleotide
sequence of the polynucleotides contained in the deposited
material, as well as the amino acid sequence of the polypeptide
encoded thereby, are controlling in the event of any conflict with
any description of sequences herein.
[0040] "Antibodies" as used herein includes polyclonal and
monoclonal antibodies, chimeric, single chain, and humanized
antibodies, as well as Fab fragments, including the products of an
Fab or other immunoglobulin expression library.
[0041] "Isolated" means altered "by the hand of man" from the
natural state. If an "isolated" composition or substance occurs in
nature, it has been changed or removed from its original
environment, or both. For example, a polynucleotide or a
polypeptide naturally present in a living animal is not "isolated,"
but the same polynucleotide or polypeptide separated from the
coexisting materials of its natural state is "isolated", as the
term is employed herein.
[0042] "Polynucleotide" generally refers to any polyribonucleotide
or polydeoxyribonucleotide, which may be unmodified RNA or DNA or
modified RNA or DNA. "Polynucleotides" include, without limitation
single- and double-stranded DNA, DNA that is a mixture of single-
and double-stranded regions, single- and double-stranded RNA, and
RNA that is mixture of single- and double-stranded regions, hybrid
molecules comprising DNA and RNA that may be single-stranded or,
more typically, double-stranded or a mixture of single- and
double-stranded regions. In addition, "polynucleotide" refers to
triple-stranded regions comprising RNA or DNA or both RNA and DNA.
The term polynucleotide also includes DNAs or RNAs containing one
or more modified bases and DNAs or RNAs with backbones modified for
stability or for other reasons. "Modified" bases include, for
example, tritylated bases and unusual bases such as inosine. A
variety of modifications has been made to DNA and RNA; thus,
"polynucleotide" embraces chemically, enzymatically or
metabolically modified forms of polynucleotides as typically found
in nature, as well as the chemical forms of DNA and RNA
characteristic of viruses and cells. "Polynucleotide" also embraces
relatively short polynucleotides, often referred to as
oligonucleotides.
[0043] "Polypeptide" refers to any peptide or protein comprising
two or more amino acids joined to each other by peptide bonds or
modified peptide bonds, i.e., peptide isosteres. "Polypeptide"
refers to both short chains, commonly referred to as peptides,
oligopeptides or oligomers, and to longer chains, generally
referred to as proteins. Polypeptides may contain amino acids other
than the 20 gene-encoded amino acids. "Polypeptides" include amino
acid sequences modified either by natural processes, such as
posttranslational processing, or by chemical modification
techniques which are well known in the art. Such modifications are
well described in basic texts and in more detailed monographs, as
well as in a voluminous research literature. Modifications can
occur anywhere in a polypeptide, including the peptide backbone,
the amino acid side-chains and the amino or carboxyl termini. It
will be appreciated that the same type of modification may be
present in the same or varying degrees at several sites in a given
polypeptide. Also, a given polypeptide may contain many types of
modifications. Polypeptides may be branched as a result of
ubiquitination, and they may be cyclic, with or without branching.
Cyclic, branched and branched cyclic polypeptides may result from
posttranslation natural processes or may be made by synthetic
methods. Modifications include acetylation, acylation,
ADP-ribosylation, amidation, covalent attachment of flavin,
covalent attachment of a heme moiety, covalent attachment of a
nucleotide or nucleotide derivative, covalent attachment of a lipid
or lipid derivative, covalent attachment of phosphotidylinositol,
cross-linking, cyclization, disulfide bond formation,
demethylation, formation of covalent cross-links, formation of
cystine, formation of pyroglutamate, formylation,
gamma-carboxylation, glycosylation, GPI anchor formation,
hydroxylation, iodination, methylation, myristoylation, oxidation,
proteolytic processing, phosphorylation, prenylation,
raceemization, selenoylation, sulfation, transfer-RNA mediated
addition of amino acids to proteins such as arginylation, and
ubiquitination. (See, for instance, PROTEINS--STRUCTURE AND
MOLECULAR PROPERTIES, 2nd Ed., T. E. Creighton, W. H. Freeman and
Company, New York, 1993 and Wold, F., Posttranslational Protein
Modifications: Perspectives and Prospects, pgs. 1-12 in
POSTTRANSLATIONAL COVALENT MODIFICATION OF PROTEINS, B. C. Johnson,
Ed., Academic Press, New York, 1983; Seifter et al., "Analysis for
protein modifications and nonprotein cofactors", Meth Enzymol
(1990) 182:626-646 and Rattan et al., "Protein Synthesis:
Posttranslational Modifications and Aging", Ann NY Acad Sci (1992)
663:48-62.)
[0044] "Variant" as the term is used herein, is a polynucleotide or
polypeptide that differs from a reference polynucleotide or
polypeptide respectively, but retains essential properties. A
typical variant of a polynucleotide differs in nucleotide sequence
from another, reference polynucleotide. Changes in the nucleotide
sequence of the variant may or may not alter the amino acid
sequence of a polypeptide encoded by the reference polynucleotide.
Nucleotide changes may result in amino acid substitutions,
additions, deletions, fusions and truncations in the polypeptide
encoded by the reference sequence, as discussed below. A typical
variant of a polypeptide differs in amino acid sequence from
another, reference polypeptide. Generally, differences are limited
so that the sequences of the reference polypeptide and the variant
are closely similar overall and, in many regions, identical. A
variant and reference polypeptide may differ in amino acid sequence
by one or more substitutions, additions, deletions in any
combination. A substituted or inserted amino acid residue may or
may not be one encoded by the genetic code. A variant of a
polynucleotide or polypeptide may be a naturally occurring such as
an allelic variant, or it may be a variant that is not known to
occur naturally. Non-naturally occurring variants of
polynucleotides and polypeptides may be made by mutagenesis
techniques or by direct synthesis.
[0045] "Identity" is a measure of the identity of nucleotide
sequences or amino acid sequences. In general, the sequences are
aligned so that the highest order match is obtained. "Identity" per
se has an art-recognized meaning and can be calculated using
published techniques. (See, e.g.: COMPUTATIONAL MOLECULAR BIOLOGY,
Lesk, A. M., ed., Oxford University Press, New York, 1988;
BIOCOMPUTING: INFORMATICS AND GENOME PROJECTS, Smith, D. W., ed.,
Academic Press, New York, 1993; COMPUTER ANALYSIS OF SEQUENCE DATA,
PART I, Griffin, A. M., and Griffin, H. G., eds., Humana Press, New
Jersey, 1994; SEQUENCE ANALYSIS 1N MOLECULAR BIOLOGY, von Heinje,
G., Academic Press, 1987; and SEQUENCE ANALYSIS PRIMER, Gribskov,
M. and Devereux, J., eds., M Stockton Press, New York, 1991.) While
there exist a number of methods to measure identity between two
polynucleotide or polypeptide sequences, the term "identity" is
well known to skilled artisans. (Carillo, H., and Lipton, D., SIAM
J Applied Math (1988) 48:1073.) Methods commonly employed to
determine identity or similarity between two sequences include, but
are not limited to, those disclosed in Guide to Huge Computers,
Martin J. Bishop, ed., Academic Press, San Diego, 1994, and
Carillo, H., and Lipton, D., SIAM J Applied Math (1988) 48:1073.
Methods to determine identity and similarity are codified in
computer programs. Preferred computer program methods to determine
identity and similarity between two sequences include, but are not
limited to, GCS program package (Devereux, J., et al., Nucleic
Acids Research (1984) 12(1):387), BLASTP, BLASTN, FASTA (Atschul,
S. F. et al., J Molec Biol (1990) 215:403.)
[0046] As an illustration, by a polynucleotide having a nucleotide
sequence having at least, for example, 95% "identity" to a
reference nucleotide sequence of SEQ ID NO:X is intended that the
nucleotide sequence of the polynucleotide is identical to the
reference sequence except that the polynucleotide sequence may
include up to five point mutations per each 100 nucleotides of the
reference nucleotide sequence of SEQ ID NO: X. In other words, to
obtain a polynucleotide having a nucleotide sequence at least 95%
identical to a reference nucleotide sequence, up to 5% of the
nucleotides in the reference sequence may be deleted or substituted
with another nucleotide, or a number of nucleotides up to 5% of the
total nucleotides in the reference sequence may be inserted into
the reference sequence. These mutations of the reference sequence
may occur at the 5' or 3' terminal positions of the reference
nucleotide sequence or anywhere between those terminal positions,
interspersed either individually among nucleotides in the reference
sequence or in one or more contiguous groups within the reference
sequence.
[0047] Similarly, by a polypeptide having an amino acid sequence
having at least, for example, 95% "identity" to a reference amino
acid sequence of SEQ ID NO:Y is intended that the amino acid
sequence of the polypeptide is identical to the reference sequence
except that the polypeptide sequence may include up to five amino
acid alterations per each 100 amino acids of the reference amino
acid of SEQ ID NO:Y. In other words, to obtain a polypeptide having
an amino acid sequence at least 95% identical to a reference amino
acid sequence, up to 5% of the amino acid residues in the reference
sequence may be deleted or substituted with another amino acid, or
a number of amino acids up to 5% of the total amino acid residues
in the reference sequence may be inserted into the reference
sequence. These alterations of the reference sequence may occur at
the amino or carboxy terminal positions of the reference amino acid
sequence or anywhere between those terminal positions, interspersed
either individually among residues in the reference sequence or in
one or more contiguous groups within the reference sequence.
Polypeptides of the Invention
[0048] In one aspect, the present invention relates to receptor
polypeptides (or receptor proteins). The receptor polypeptides
include the polypeptide of SEQ ID NO:Y; as well as polypeptides
comprising the amino acid sequence of SEQ ID NO:Y; and polypeptides
comprising the amino acid sequence which have at least 80% identity
to that of SEQ ID NO:Y over its entire length, and still more
preferably at least 90% identity, and even still more preferably at
least 95% identity to SEQ ID NO:Y. Furthermore, those with at least
97-99% identity to SEQ ID NO:Y are highly preferred. Also included
within receptor polypeptides are polypeptides having the amino acid
sequence which have at least 80% identity to the polypeptide having
the amino acid sequence of SEQ ID NO:Y over its entire length, and
still more preferably at least 90% identity, and even still more
preferably at least 95% identity to SEQ ID NO:Y. Furthermore, those
with at least 97-99% are highly preferred. Preferably receptor
polypeptides exhibit at least one biological activity of the
receptor.
[0049] The receptor polypeptides may be in the form of the "mature"
protein or may be a part of a larger protein such as a fusion
protein. It is often advantageous to include an additional amino
acid sequence which contains secretory or leader sequences,
pro-sequences, sequences which aid in purification such as multiple
histidine residues, or an additional sequence for stability during
recombinant production.
[0050] Fragments of the receptor polypeptides are also included in
the invention. A "fragment" is a polypeptide having an amino acid
sequence that entirely is the same as part, but not all, of the
amino acid sequence of the aforementioned receptor polypeptides. As
with receptor polypeptides, fragments may be "free-standing," or
comprised within a larger polypeptide of which they form a part or
region, most preferably as a single continuous region.
Representative examples of polypeptide fragments of the invention,
include, for example, fragments from about amino acid number 1-20,
21-40, 41-60, 61-80, 81-100, and 101 to the end of receptor
polypeptide. In this context "about" includes the particularly
recited ranges larger or smaller by several, 5, 4, 3, 2 or 1 amino
acid at either extreme or at both extremes.
[0051] Preferred fragments include, for example, truncation
polypeptides having the amino acid sequence of receptor
polypeptides, except for deletion of a continuous series of
residues that includes the amino terminus, or a continuous series
of residues that includes the carboxyl terminus or deletion of two
continuous series of residues, one including the amino terminus and
one including the carboxyl terminus.
[0052] Also preferred are fragments characterized by structural or
functional domains, such as fragments that comprise alpha-helix and
alpha-helix forming regions, beta-sheet and beta-sheet-forming
regions, turn and turn-forming regions, coil and coil-forming
regions, hydrophilic regions, hydrophobic regions, alpha
amphipathic regions, beta amphipathic regions, flexible regions,
surface-forming regions, substrate binding region, and high
antigenic index regions. The "domains" of each receptor polypeptide
are illustrated in the Figures. The Figures compare SEQ ID NO:Y to
the closest know homologue. Identical amino acids shared between
the two polypeptides are shaded, while conservative amino acid
changes are boxed. By examining the regions or amino acids shaded
and/or boxed, the skilled artisan can readily identify conserved
domains between the two polypeptides. The amino acids sequences of
SEQ ID NO:Y falling within these conserved domains are "fragments"
and are specifically contemplated by the present invention.
Especially preferred is the extracellular domains of a receptor of
the invention. Soluble extracellular domains have antagonist
activity mediated by competition with a receptor ligand.
[0053] Other preferred fragments are biologically active fragments.
Biologically active fragments are those that mediate receptor
activity, including those with a similar activity or an improved
activity, or with a decreased undesirable activity. Also included
are those that are antigenic or immunogenic in an animal,
especially in a human.
[0054] Preferably, all of these polypeptide fragments retain a
biological activity of the receptor, including antigenic activity.
Variants of the defined sequence and fragments also form part of
the present invention. Preferred variants are those that vary from
the referents by conservative amino acid substitutions--i.e., those
that substitute a residue with another of like characteristics.
Typical such substitutions are among Ala, Val, Leu and Ile; among
Ser and Thr; among the acidic residues Asp and Glu; among Asn and
Gln; and among the basic residues Lys and Arg; or aromatic residues
Phe and Tyr. Particularly preferred are variants in which several,
5-10, 1-5, or 1-2 amino acids are substituted, deleted, or added in
any combination.
[0055] The receptor polypeptides of the invention can be prepared
in any suitable manner. Such polypeptides include isolated
naturally occurring polypeptides, recombinantly produced
polypeptides, synthetically produced polypeptides, or polypeptides
produced by a combination of these methods. Means for preparing
such polypeptides are well understood in the art.
Polynucleotides of the Invention
[0056] Another aspect of the invention relates to receptor
polynucleotides. Receptor polynucleotides include isolated
polynucleotides which encode the receptor polypeptides and
fragments, and polynucleotides closely related thereto. More
specifically, a receptor polynucleotide of the invention includes a
polynucleotide comprising the nucleotide sequence contained in SEQ
ID NO:X encoding a receptor polypeptide of SEQ ID NO:Y, and
polynucleotide having the particular sequence of SEQ ID NO:X.
[0057] Receptor polynucleotides further include a polynucleotide
comprising a nucleotide sequence that has at least 80% identity
over its entire length to a nucleotide sequence encoding the
receptor polypeptide of SEQ ID NO:Y, and a polynucleotide
comprising a nucleotide sequence that is at least 80% identical to
that of SEQ ID NO:X over its entire length. In this regard,
polynucleotides at least 90% identical are particularly preferred,
and those with at least 95% are especially preferred. Furthermore,
those with at least 97% are highly preferred and those with at
least 98-99% are most highly preferred, with at least 99% being the
most preferred. Also included under receptor polynucleotides are a
nucleotide sequence which has sufficient identity to a nucleotide
sequence contained in SEQ ID NO:X, or contained in the cDNA insert
in the plasmid deposited with ATCC, to hybridize under conditions
useable for amplification or for use as a probe or marker.
Moreover, the receptor polynucleotide includes a nucleotide
sequence having at least 80% identity to a nucleotide sequence
encoding the receptor polypeptide expressed by the cDNA insert
deposited at the ATCC, and a nucleotide sequence comprising at
least 15 contiguous nucleotides of such cDNA insert. In this
regard, polynucleotides at least 90% identical are particularly
preferred, and those with at least 95% are especially preferred.
Furthermore, those with at least 97% are highly preferred and those
with at least 98-99% are most highly preferred, with at least 99%
being the most preferred. The invention also provides
polynucleotides which are complementary to all the above receptor
polynucleotides.
[0058] The receptors of the invention are structurally related to
other proteins of specified receptor families, as shown by the
results in the Figures. The cDNA sequence of SEQ ID NO:X encodes a
polypeptide as described in Table 1 as SEQ ID NO:Y. Because the
receptor polypeptides contain domains similar in structure to other
receptor family members, the receptors of the present invention are
expected to have, inter alia, similar biological
functions/properties to their homologous polypeptides and
polynucleotides, and their utility is obvious to anyone skilled in
the art. TABLE-US-00001 TABLE 1 SEQ ID SEQ ID ATCC ATCC Receptor
Clone ID Name NO: X NO: Y Deposit No. Deposit Date Family Homology
HMACR70 1 18 209054 May 16, 1997 Ig Sialoadhesin ##### Jan. 21,
1998 OB-1 HTEDK48 209054 May 16, 1997 TM4SF MRC-OX44 PETA-3 1-1849
bp 2 160-900 bp 3 19 HTPED39 4 20 209054 May 16, 1997 TM4SF NAG-2
HPWAE25 ##### Jan. 21, 1998 TALLA-1 HTPEF86 5 21 209053 May 16,
1997 TM4SF CD20 B1 Antigen HSBBF02 6 22 209054 May 16, 1997 TM4SF
TALLA-1 HLTAH80 7 23 97242 Aug. 02, 1995 TM4SF TALLA-1 209054 May
16, 1997 HTPBA27 8 24 97242 Aug. 02, 1995 TM4SF NAG-2 209054 May
16, 1997 HAIDQ59 209054 May 16, 1997 TM4SF CD9 Antigen 5' Sequence
9 25 3' Sequence 10 HHFEK40 11 26 209054 May 16, 1997 TM4SF PETA-3
HGBGV89 12 27 209125 Jun. 09, 1997 TM4SF L6H 209054 May 16, 1997
HUVBB80 13 28 209054 May 16, 1997 TM4SF L6 HJACE54 14 29 209053 May
16, 1997 Lectin Galectin-3 Galectin-5 Galectin-8 HROAD63 15 30
209053 May 16, 1997 Ly6 E48 splice variant HMWGS46 16 31 209053 May
16, 1997 Prohibitin BAP-37 HNFGW06 17 32 209053 May 16, 1997 EGFR
EGFR
[0059] The novel full-length cDNA clone designated HMACR70 (SEQ ID
NO:1) may be a member of the sialoadhesin family of the Ig
superfamily of receptor-like molecules and a CD33 homologue.
HMACR70 contains a 1497 nucleotide cDNA insert (SEQ ID NO:1)
encoding a 315 amino acid ORF (SEQ ID NO:18) and was cloned from a
GM-CSF-treated human macrophage cDNA library. The only additional
cDNA libraries in the HGS database which include this clone are
human eosinophils and possibly human gall bladder. A BLAST analysis
of the amino acid sequence of HMACR70 (SEQ ID NO:18) demonstrates
that this clone exhibits approximately 50% identity and 69%
similarity over a 300 amino acids stretch of a gene termed human
differentiation antigen, and 38% identity and 62% similarity of the
human myelin-associated glycoprotein precursor CD33 gene.
[0060] A more recent BLAST analysis confirms HMACR70's (SEQ ID
NO:18) designation as a sialoadhesin family member. HMACR70 (SEQ ID
NO:18) is homologous to two recently identified sialoadhesin family
members, human OB binding protein (OB) 1 (SEQ ID NO:33) and 2.
(See, Genbank Accession No. U71382; see FIG. 1.) It is thought that
OB-1 (SEQ ID NO:33) and OB-2 may bind leptin. Thus, HNACR70 (SEQ ID
NO:18), as a sialoadhesin family member, may act to attenuate or
even amplify intercellular routes of communication, including
binding to leptin or modulating the activity of immune cells, such
as macrophages. Clearly, any diseases affected by these processes
could be treated by the polypeptide or fragment of HMACR70 (SEQ ID
NO:18).
[0061] The full-length nucleotide sequences of ten novel human cDNA
clones which potentially belong to the TM4SF superfamily are
disclosed in the table above and will be addressed
sequentially.
[0062] The cDNA clone HTEDK48 contains a 1849 nucleotide cDNA
insert (SEQ ID NO:2) encoding a 245 amino acid ORF that was cloned
from a human testes cDNA library. The coding sequence of HTEDK48
(SEQ ID NO: 3) may be fused to other human proteins, such as
3-hydroxyacyl-CoA dehydrogenase. BLAST analysis of the amino acid
sequence of HTEDK48 (SEQ ID NO:19) demonstrates that this clone
exhibits approximately 30% identity and 51% similarity over a 245
amino acid stretch of the CD82 molecule. Recent studies have shown
that CD82 can associate with CD4 or CD8 and deliver costimulatory
signals for the TCR/CD3 pathway. CD82 has also been found to be
involved in syncytium formation in HTLV-1-infected T-cells. And
finally, in a recently published study in which the expression of
the CD82 gene by tumors of the lung was examined retrospectively,
it was reported that CD82 may be linked to the suppression of tumor
metastasis of prostate cancer. The study also reported that
decreased CD82 expression may be involved in malignant progression
of such cancers. Thus, HTEDK48 (SEQ ID NO:2, NO:3, NO:19) may also
be involved in the development of cancer.
[0063] A more recent BLAST analysis shows that HTEDK48 (SEQ ID
NO:19) is homologous the rat leukocyte antigen, MRC OX-44 (SEQ ID
NO:34), and the platelet endothelial tetraspan antigen-3 (PETA-3)
(SEQ ID NO:35). (See FIG. 2X.) MRC OX-44 (SEQ ID NO:34), a member
of a new family of cell surface proteins, appears to be involved in
growth regulation. (See, Bellacosa, A., et al., "The Rat Leukocyte
antigen MRC OX-44 is a Member of a New Family of Cell Surface
Proteins which Appear to be Involved in Growth Regulation," Mol.
Cell. Bio. 11: 2864-2872 (1991).) Similarly, PETA-3 (SEQ ID NO:35)
has been located to platelet endothelial cells, and an anti-PETA-3
antigen monoclonal antibody can stimulate platelet aggregation and
mediator release. (See, Fitter, S., "Molecular Cloning of cDNA
Encoding a Novel Platelet-Endothelial Cell Tetra-Span Antigen,
PETA-3," Blood, 86(4):1348-1355 (1995).) Thus, HTEDK48 (SEQ ID
NO:19) may function similar to MRC OX-44 (SEQ ID NO:34) or PETA-3
(SEQ ID NO:35) to affect growth of blood cells. Administering
polypeptides or fragments of HTEDK48 (SEQ ID NO:19) may be an
effective treatment of blood disorders.
[0064] The cDNA clone HPWAE25 contains a 1288 nucleotide cDNA
insert (SEQ ID NO:4) encoding a 273 amino acid ORF (SEQ ID NO:20)
that was cloned from a human pancreas tumor cDNA library, while
clone HTPED39 represents a truncated cDNA sequence. This clone also
appears in a number of other cDNA libraries constructed from a
variety of human cell and tissue types including keratinocytes,
ulcerative colitis, striatum depression, lymph node breast cancer,
ovarian cancer, stage B2 prostate cancer, kidney medulla, and
others. Northern blot analysis of HLTAH80 (SEQ ID NO:23) also shows
expression in a variety of human cell lines including U937, MM96,
WM115, and MDAMB231. A BLAST analysis of the amino acid sequence of
HTPED39 demonstrates that this clone exhibits approximately 35%
identity and 50% similarity over the entire length of the CD37
molecule. The CD37 antigen is expressed on B cells and on a
subpopulation of T cells, but not on pre-B or plasma cells. It has
been reported that CD37 expression is downregulated in conjunction
with B-cell activation, suggesting that CD37 may be involved in the
processes which dictate the activation state of the B-cell.
[0065] Moreover, HPWAE25 (SEQ ID NO:20) is also homologous to
recently identified TM4SF members, NAG-2 (SEQ ID NO:36) and TALLA-1
(SEQ ID NO:37). (See FIG. 3.) NAG-2 (SEQ ID NO:36) is thought to
complex with integrins and other TM4SF proteins, while TALLA-1 (SEQ
ID NO:37) is a highly specific marker of T-cell acute lymphoblastic
leukemia and neuroblastoma. (See, Tachibana, I., et al., "NAG-2, A
Novel Transmembrane-4 Superfamily (TM4SF) Protein that Complexs
with Integrins and Other TM4SF Proteins," J. Biol. Chem.,
272:29181-29189 (1997); Takagi, S., "Identification of a Highly
Specific Surface Marker of T-cell Acute Lymphoblastic Leukemia and
Neuroblastoma as a New Member of the Transmembrane 4 Superfamily,"
Int. J. Cancer 61(5):706-715 (1995).) Thus, HPWAE25 (SEQ ID NO:20)
may be involved the development of cancer, particularly leukemia,
lymphoma, and neuroblastoma. HPWAE25 (SEQ ID NO:4 and NO:20) may be
used as an effective treatment of these cancers, as well as a
diagnostic marker.
[0066] A subfamily of TM4SF receptors include CD20 proteins. A
CD20-like cDNA clone was obtained from a human pancreas tumor cDNA
library and contains a 1236 nucleotide insert which encodes a 250
amino acid ORF. A BLAST analysis of the deduced amino acid sequence
of HTPEF86 (SEQ ID NO:21) exhibits approximately 41% identity and
61% similarity to the CD20 gene, also known as B1 antigen (SEQ ID
NO:38). (See FIG. 4.) Expression of this gene is detected in only
two additional HGS human cDNA libraries; amygdala depression and 9
week early stage human. Although the precise functional role of
CD20 has yet to be determined, it is clear that CD20 plays a key
role in the regulation of B-cell activation. Based primarily on
sequence identity, the novel CD20-like molecule presented herein
may also be involved in cell cycle activation. Potential
therapeutic and/or diagnostic applications for HTPEF86 (SEQ ID
NO:21) may include such clinical presentations as juvenile
rheumatoid arthritis, Graves' Disease, and a number of B-cell
lymphomas or other lymphoid tumors.
[0067] The clone HSBBF02 contains a 1115 nucleotide cDNA insert
(SEQ ID NO:6) encoding a 245 amino acid ORF (SEQ ID NO:22) and was
cloned from an HSC 172 cell line cDNA library. This clone also
appears in a number of other cDNA libraries constructed from a
variety of human cell and tissue types including brain amygdala
depression, endothelial cells, fetal liver and heart, osteoblasts,
testes, and others. A BLAST analysis of the amino acid sequence of
HSBBF02 (SEQ ID NO:22) demonstrates that this clone exhibits
approximately 64% identity and 80% similarity with the A15 molecule
over a 131 amino acid stretch (A15 is composed of 244 amino acids).
A more recent BLAST search shows that HSBBF02 (SEQ ID NO:22) is
similar to the TALLA-1 protein (SEQ ID NO:37) and may in fact be a
closely related family member. (See FIG. 5.)
[0068] In addition, a second cDNA clone, designated HLTAH80 (SEQ ID
NO:23), exhibits sequence similarity to the A15 molecule and
TALLA-1 (SEQ ID NO:37). (See FIG. 6.) This clone contains a 1662
nucleotide cDNA insert encoding a 253 amino acid ORF and was cloned
from a human T-cell lymphoma cDNA library. This clone also appears
in a number of other cDNA libraries constructed from a variety of
human cell and tissue types including B-cell lymphoma, corpus
collosum, endometrial tumor, osteosarcoma, testes, and others.
Northern blot analysis of HLTAH80 (SEQ ID NO:7) also shows
expression in a variety of human tissues including spleen, lymph
node, thymus, PBLs, heart, and a particularly strong signal in
skeletal muscle and pancreas. A BLAST analysis of the amino acid
sequence of HLTAH80 (SEQ ID NO:23) demonstrates that this clone
exhibits approximately 35% identity and 55% similarity over the
entire length of the A15 molecule.
[0069] Since expression of A15 drops to undetectable levels when
comparing immature T-cells to peripheral blood lymphocytes, it is
thought that A15 may play a role in the development of T-cells.
Furthermore, the MXS1 (CCG-B7) gene which codes for A15 contains a
number of triplet nucleotide repeats which have been associated
with neuropsychiatric diseases such as Huntington's chorea, fragile
X syndrome, and myotonic dystrophy. In addition, A15 appears to be
expressed exclusively on T-cell acute lymphoblastic leukemia cell
lines, including several derived from adult T-cell leukemia and
those established by immortalization with human T-cell leukemia
virus type 1 or Herpesvirus saimiri. Thus, clones HLTAH80 (SEQ ID
NO:7 and NO:23) and/or HSBBF02 (SEQ ID NO:6 and NO:22) may also be
involved in diseases caused by the expansion of repeats or
chromosomal instability.
[0070] The cDNA clone HTPBA27 contains a 1345 nucleotide cDNA
insert (SEQ ID NO:8) encoding a 238 amino acid ORF (SEQ ID NO:24)
and was cloned from a human tumor pancreas cDNA library. This clone
also appears in a number of other cDNA libraries constructed from a
variety of human cell and tissue types including cerebellum, breast
lymph node, osteosarcoma, adult testes, RS4; 11 bone marrow cell
line, microvascular endothelial cells, and others. A BLAST analysis
of the amino acid sequence of HTPBA27 (SEQ ID NO:24) demonstrates
that this clone exhibits approximately 40% identity and 64%
similarity with a glycoprotein termed CD53 over its entire length.
CD53 is thought to be involved in thymopoiesis, since rat CD53 can
be detected on immature CD4-8-thymocytes and the functionally
mature single-positive subset, but cannot be detected on the
intermediate CD4+8+thymocytic subset of cells. The CD53 molecule
has also been implicated as a component of signal transduction
pathways in B cells, monocytes and granulocytes, rat macrophages,
NK, and T cells. Moreover, as illustrated in FIG. 7, HTPBA27 (SEQ
ID NO:24) was recently confirmed as a TM4SF receptor. (See,
Tachibana, I., et al., "NAG-2, A Novel Transmembrane-4 Superfamily
(TM4SF) Protein that with Integrins and Other TM4SF Proteins," J.
Biol. Chem., 272:29181-29189 (1997).) Calling the HTPBA27
polypeptide (SEQ ID NO:24) NAG-2 (SEQ ID NO:36), this group
confirmed HTPBA27's status as a TM4SF receptor by showing that
NAG-2 (SEQ ID NO:36) complexes with integrin and other TM4SF
receptors. Thus, diseases caused by the failure of HTPBA27 (SEQ ID
NO:24) to complex with integrin and other TM4SF receptors can be
treated by administering HTPBA27 (SEQ ID NO:24). HTPBA27 (SEQ ID
NO:8 and NO:24) can also be used to diagnose these diseases.
[0071] The cDNA clone HAIDQ59 contains cDNA insert encoding a 221
amino acid ORF (SEQ ID NO:25) that was cloned from a human
epithelial cell induced with TNFa and INF cDNA library. The 5' end
of HAIDQ59 is represented by the SEQ ID NO: 9, while the 3' end is
represented by SEQ ID NO: 10. This clone appears in only two
additional cDNA libraries in the HGS database. These two libraries
were constructed from the human Jurkat T-cell line and human
microvascular endothelial cells. A BLAST analysis of the amino acid
sequence of HAIDQ59 (SEQ ID NO:25) demonstrates that this clone
exhibits approximately 53% identity and 69% similarity over 226
amino acids of the CD9 TM4SF molecule (SEQ ID NO:39). (See FIG. 8.)
It has been demonstrated that the CD9 molecule (SEQ ID NO:39) is
involved in signal transduction pathways in platelets, as well as
in cell adhesion in both platelets and pre-B-cell lines.
Intriguingly, a monoclonal antibody (vpg15), which recognizes the
feline homologue of CD9, has been shown to block infection by
feline immunodeficiency virus (FIV). Furthermore, a recent study
shows that cells expressing high levels of CD9 (SEQ ID NO:39)
exhibited suppressed cell motility. Thus, HAIDQ59 (SEQ ID NO:25)
may also be involved in signal transduction of blood cells.
[0072] The cDNA clone HHFEK40 contains a 936 nucleotide cDNA insert
(SEQ ID NO:11) encoding a 252 amino acid ORF (SEQ ID NO:26) and was
cloned from a human fetal heart cDNA library. This clone appears
once in the human fetal heart cDNA library and possibly in a
hemangiopericytoma cDNA library. A BLAST analysis of the amino acid
sequence of HHFEK40 (SEQ ID NO:26) demonstrated that this clone
exhibits approximately 60% identity and 75% similarity over the
entire length of a molecule designated PETA-3 (SEQ ID NO:35). (See
FIG. 9.) PETA-3 (SEQ ID NO:35) was originally identified as a novel
human platelet surface glycoprotein termed gp27. Although PETA-3
(SEQ ID NO:35) is present in low abundance on the platelet surface,
an anti-PETA-3 monoclonal antibody can stimulate platelet
aggregation and mediator release. Thus, HHFEK40 (SEQ ID NO:26) may
function similar to PETA-3 (SEQ ID NO:35) to affect growth of blood
cells. Administering polypeptides or fragments of HHFEK40 (SEQ ID
NO:26) may be an effective treatment of blood disorders.
[0073] The cDNA clone HGBGV89 contains a 738 nucleotide cDNA insert
(SEQ ID NO:12) encoding a 197 amino acid ORF (SEQ ID NO:27) and was
cloned from a human gall bladder cDNA library. The only two
additional appearances of this clone in the HGS database are in a
normalized fetal liver cDNA library and in a fetal liver/spleen
cDNA library. The cDNA clone HUVBB80 contains a 1071 nucleotide
cDNA insert (SEQ ID NO:13) encoding a 201 amino acid ORF (SEQ ID
NO:28) and was cloned from a human umbilical vein cDNA library.
This clone appears in several additional cDNA libraries in the HGS
database including prostate BPH, thyroid, and fetal liver/spleen.
BLAST analyses of the amino acid sequences of HGBGV89 (SEQ ID
NO:27) and HUVBB80 (SEQ ID NO:28) demonstrate that these clones
exhibit approximately 49% identity and 65% similarity and 47%
identity and 68% similarity, respectively, over the entire length
of a molecule designated L6 surface protein (SEQ ID NO:41) or human
tumor-associated antigen L6 (SEQ ID NO:41) (See FIGS. 10 & 11.)
Moreover, another group has confirmed the TM4SF receptor homology
of HGBGV89 (SEQ ID NO:27) by describing the protein as a putative
transmembrane protein L6H (SEQ ID NO:40). (See Genbank Accession No
2587054; see FIG. 10.) The L6 cell surface antigen (SEQ ID NO:41)
is highly expressed on lung, breast, colon, and ovarian carcinomas.
Promising results of phase 1 clinical studies have been reported
with an anti-L6 monoclonal antibody, or its humanized counterpart,
suggesting that the L6 antigen (SEQ ID NO:41) may be an attractive
target for monoclonal antibody-based cancer therapy.
[0074] In summary, there is a clear need for identifying and
exploiting novel members of the TM4SF superfamily such as those
described herein. Although structurally related, these factors will
likely possess diverse and multifaceted functions in a variety of
cell and tissue types. Receptor type molecules, such as the novel
potential members of the TM4SF superfamily detailed here, should
prove useful in target based screens for small molecules and other
such pharmacologically valuable factors. Monoclonal antibodies
raised against such factors may prove useful as therapeutics in an
anti-tumor, diagnostic, or other capacity. Furthermore, factors
such as the nine novel TM4SF superfamily-like molecules described
here may prove useful in an active or passive immunotherapeutical
role in patients with cancer or other immunocompromised disease
states.
[0075] Besides TM4SF receptors, receptors from other families are
also described. For example, clone HJACE54 (SEQ ID NO:14 and
NO:29), also called galectin 11, exhibits significant sequence
identity to the rat galectin 5 (SEQ ID NO:42), the chicken galectin
3 gene, and the human galectin 8 (SEQ ID NO:43) genes. (See FIG.
12.) The galectin 11 cDNA clone contains an 865 nucleotide insert
(SEQ ID NO:14) which encodes a 133 amino acid ORF (SEQ ID NO:29).
The clone was obtained from a Jurkat T-cell G1 phase cDNA library.
A BLAST analysis of the deduced amino acid sequence of HJACE54 (SEQ
ID NO:29) demonstrates approximately 35% identity and 57%
similarity to the amino acid sequence of the rat galectin 5 (SEQ ID
NO:42) gene. Expression of galectin 11 (SEQ ID NO:14) is quite
limited in the HGS database. In fact, the only two additional ESTs
in the HGS database which contain the HJACE54 sequence (SEQ ID
NO:14) were found in human neutrophil and human infant adrenal
gland cDNA libraries. Northern blot analyses have not been
performed to examine expression patterns of the galectin 11 gene
(SEQ ID NO:14).
[0076] Various galectins have been shown to function in the
mechanisms of intercellular communication. For example, depending
on cell type, galectin 1 has been observed to modulate cell
adhesion either positively or negatively. More specifically,
galectin 1 appears to inhibit cell adhesion of skeletal muscle
presumably by galectin 1-mediated disruption of laminin-integrin
a.sub.7b.sub.1 interactions. Alternatively, galectin 1 appears to
promote cell adhesion in several non-skeletal muscle cell types
examined presumably by a glycoconjugate cross-linking mechanism.
Galectin 3 has also been observed to function in modulating
cell-adhesion, as well as in the activation of certain immune cells
by cross-linking IgE and IgE receptors. In addition, galectins have
been observed to be involved in the regulation of immune cell
activity, as well as in such diverse processes as cell adhesion,
proliferation, inflammation, autoimmunity, and metastasis of tumor
cells. Furthermore, a galectin-like antigen designated HOM-HD-21
was recently found to be highly expressed in a Hodgkin's Disease
cDNA library. Very recently, a novel galectin, termed PCTA-1, was
identified as a specific cell surface marker on human prostate
cancer cell lines and patient-derived carcinomas. Galectins have
also been found to function intracellularly as a component of
ribonucleoprotein complexes. Finally, galedtins 1 and 3 have each
been found to modulate T-cell growth and apoptosis by interaction
with CD45 and possibly Bcl2, respectively. As a result, the
discovery of a novel galectin (SEQ ID NO:29), such as that encoded
by HJACE54 (SEQ ID NO:14), is likely to be a valuable asset both
diagnostically and therapeutically.
[0077] Additionally, a full-length nucleotide sequence of a novel
human cDNA clone which encodes an apparent splice variant of the
previously described human E48 antigen has recently been
determined. (See FIG. 13.) Clone HROAD63 contains a 441 nucleotide
cDNA (SEQ ID NO:15) which encodes a 70 amino acid polypeptide (SEQ
ID NO:30). This novel clone exhibits significant sequence identity
to several members of a relatively new family of cell-surface
proteins termed the Ly6 superfamily. These members include murine
and human SCA-2, rat Ly-6 (also termed ThB), and human CD59 [also
known as protectin or membrane attack complex inhibition factor
(MACIF)]. The novel E48 splice variant (SEQ ID NO:15) was obtained
from the HGS human stomach cDNA library. The clone (SEQ ID NO:30)
is present in only a limited number of other HGS cDNA libraries
including kidney cancer, keratinocyte, and tongue. An alignment of
the nucleotide sequences of the human E48 and HROAD63 (SEQ ID
NO:15) cDNAs demonstrates that the initial 168 and 178 nucleotides
of E48 and HROAD63, respectively, are identical, with the exception
of an additional 10 nucleotides of sequence at the extreme 5' end
of the HROAD63 sequence. The sequence of the two clones is also
identical for an additional 229 nucleotides including the 3' end of
the coding sequences and the entire 3' untranslated regions. The
only divergence of nucleotide sequence in this region of the clones
is the deletion of a single thymidine residue in the 3' UTR of the
E48 cDNA. The major difference between the two nucleotide sequences
is a 329 nucleotide deletion from the HROAD63 sequence. This
deletion causes a shift in the HROAD63 reading frame and
encompasses the translational stop signal used in the E48 clone. As
a result, the carboxy terminal sequence of HROAD63 (SEQ ID NO:30)
is radically altered with regard to that of E48 (SEQ ID NO:44) (as
illustrated in FIG. 13 by the obvious differences between amino
acids 56-128 of E48 and 56-70 of HROAD63 in the amino acid
alignment). The clinical presentation of disorders, including
abnormal skin and hair phenotypes, may be attributed, at least in
part, to a non-functional Ly6 superfamily member such as E48 (SEQ
ID NO:44) or HROAD63 (SEQ ID NO:30). HROAD63 (SEQ ID NO:30) may
also be involved in blood disorders, as seen with its homologues
SCA-2 and CD59.
[0078] A novel prohibitin cDNA clone (SEQ ID NO:16) presented
herein was originally identified in a human bone marrow cell line
(RS4; 11) cDNA library. The clone contains a 1066 nucleotide insert
(SEQ ID NO:16) which encodes a 299 amino acid polypeptide (SEQ ID
NO:31). BLAST and BestFit analyses of the predicted amino acid
sequence of HMWGS46 (SEQ ID NO:31) demonstrate a highly significant
sequence identity to a murine protein termed IgM B-cell receptor
associated protein (BAP)-37 (SEQ ID NO:45) (Genbank accession
number X78683). The HMWGS46 amino acid sequence (SEQ ID NO:31)
exhibits nearly perfect identity and similarity over the entire
length of the murine BAP-37 sequence (SEQ ID NO:45). (See FIG. 14.)
In addition, the full-length nucleotide sequences of HMWGS46 (SEQ
ID NO:16) and BAP-37 (SEQ ID NO:45) exhibit at least 87% identical.
The HMWGS46 clone (SEQ ID NO:16) also exhibits approximately 49%
sequence identity and 85% sequence similarity to a human gene
designated prohibitin. Finally, the HMWGS46 cDNA (SEQ ID NO:16)
appears in a substantial number of HGS human cDNA libraries in
addition to the bone marrow cell line cDNA library from which it
was cloned. Some of the cDNA libraries in which this clone appears
include keratinocytes, induced endothelial cells, activated
neutrophils, synovial sarcoma, colon carcinoma cell line, Jurkat
cell line membrane bound polysomes, epileptic frontal cortex,
primary dendritic cells, and a number of others. The novel gene
related to prohibitin and BAP-37 (SEQ ID NO:45) may prove quite
useful as a diagnostic for tumorigenesis, as well as a target for
therapeutic intervention of such an event. Thus, although the
precise functional role of the prohibitin family members are less
than clear, it is quite likely that such homologues are involved in
such complex processes as development, senescence, and tumor
suppression. Therefore a novel gene, such as HMWGS46 (SEQ ID NO:16
and NO:31), may prove quite useful as a diagnostic for
tumorigenesis, as well as a target for therapeutic intervention of
such an event.
[0079] A human cDNA clone encoding a novel epidermal growth factor
receptor (EGFR)-like molecule (SEQ ID NO:32) is also disclosed. The
novel EGFR-like cDNA clone (SEQ ID NO:17) presented herein was
originally identified in an activated human neutrophil cDNA
library. The clone contains a 704 nucleotide insert (SEQ ID NO:17)
which encodes a 168 amino acid polypeptide (SEQ ID NO:32). A BLAST
analysis of the predicted amino acid sequence of HNFGW06 (SEQ ID
NO:32) demonstrates that this novel clone exhibits approximately
85% identity and 90% similarity to a protein designated epidermal
growth factor receptor-related protein [Homo sapiens] (SEQ ID
NO:46). (See FIG. 15.) The expression profile of the HNFGW06 clone
(SEQ ID NO:17) in the HGS database indicates the existence of a
fairly highly restricted expression pattern. In addition to the
activated neutrophil library from which this clone (SEQ ID NO:17)
was obtained, it also appears in the following HGS human cDNA
libraries: synovial sarcoma, smooth muscle, placenta, and possibly
primary dendritic cells.
[0080] The novel EGFR-like cDNA clone HNFGW06 (SEQ ID NO:17) may
lead to a number of exciting possibilities for therapeutic and/or
diagnostic treatments or reagents. For example, HNFGW06 (SEQ ID
NO:17 and NO:32) may be involved in the onset of human breast
cancers as well. In addition, due to the fact that TGF-a acts
through binding to the EGFR (SEQ ID NO:46), it is possible that
HNFGW06 (SEQ ID NO:17 and NO:32) may also play a role in a variety
of gastric processes including regulation of acid secretion,
regulation of mucous cell growth, and protection against ethanol-
and aspirin-induced injury to gastric tissues.
Generating Polynucleotides
[0081] Polynucleotides of the present invention encoding a receptor
may be obtained using standard cloning and screening, from a cDNA
library derived from mRNA in cells specified in Table 1 using the
expressed sequence tag (EST) analysis (Adams, M. D., et al. Science
(1991) 252:1651-1656; Adams, M. D. et al., Nature, (1992)
355:632-634; Adams, M. D., et al., Nature (1995) 377 Supp:3-174.)
Polynucleotides of the invention can also be obtained from natural
sources such as genomic DNA libraries or can be synthesized using
well known and commercially available techniques.
[0082] The nucleotide sequence encoding a receptor polypeptide of
SEQ ID NO:Y may be identical to the polynucleotide encoding SEQ ID
NO:Y, or it may be a sequence, which as a result of the redundancy
(degeneracy) of the genetic code, also encodes the polypeptide of
SEQ ID NO:Y.
[0083] When the polynucleotides of the invention are used for the
recombinant production of a receptor polypeptide, the
polynucleotide may include the coding sequence for the mature
polypeptide or a fragment thereof, by itself; the coding sequence
for the mature polypeptide or fragment in reading frame with other
coding sequences, such as those encoding a leader or secretory
sequence, a pre-, or pro- or prepro-protein sequence, or other
fusion peptide portions. For example, a marker sequence which
facilitates purification of the fused polypeptide can be encoded.
In certain preferred embodiments of this aspect of the invention,
the marker sequence is a hexa-histidine peptide, as provided in the
pQE vector (Qiagen, Inc.) and described in Gentz et al., Proc Natl
Acad Sci USA (1989) 86:821-824, or is an HA tag. The polynucleotide
may also contain non-coding 5' and 3' sequences, such as
transcribed, non-translated sequences, splicing and polyadenylation
signals, ribosome binding sites and sequences that stabilize
mRNA.
[0084] Further preferred embodiments are polynucleotides encoding
receptor variants comprising the amino acid sequence of receptor
polypeptide of Table 1 (SEQ ID NO:Y) in which several, 5-10, 1-5,
1-3, 1-2 or 1 amino acid residues are substituted, deleted or
added, in any combination.
[0085] The present invention further relates to polynucleotides
that hybridize to the herein above-described sequences. In this
regard, the present invention especially relates to polynucleotides
which hybridize under stringent conditions to the herein
above-described polynucleotides. As herein used, the term
"stringent conditions" means hybridization will occur only if there
is at least 80%, and preferably at least 90%, and more preferably
at least 95%, yet even more preferably 97-99% identity between the
sequences.
[0086] Polynucleotides of the invention, which are identical or
sufficiently identical to a nucleotide sequence contained in SEQ ID
NO:X or a fragment thereof, or to the cDNA insert in the plasmid
deposited at the ATCC, or a fragment thereof, may be used as
hybridization probes for cDNA and genomic DNA, to isolate
full-length cDNAs and genomic clones encoding the receptor and to
isolate cDNA and genomic clones of other genes (including genes
encoding homologs and orthologs) that have a high sequence
similarity to the receptor gene. Such hybridization techniques are
known to those of skill in the art. Typically these nucleotide
sequences are 80% identical, preferably 90% identical, more
preferably 95% identical to that of the referent. The probes
generally will comprise at least 15 nucleotides. Preferably, such
probes will have at least 30 nucleotides and may have at least 50
nucleotides. Particularly preferred probes will range between 30
and 50 nucleotides.
[0087] In one embodiment, to obtain a polynucleotide encoding the
receptor polypeptide, including homologs and orthologs from other
species, comprises the steps of screening an appropriate library
under stringent hybridization conditions with a labeled probe
having the SEQ ID NO:X or a fragment thereof; and isolating
full-length cDNA and genomic clones containing said polynucleotide
sequence. Such hybridization techniques are well known to those of
skill in the art. Stringent hybridization conditions are as defined
above or, alternatively, conditions under overnight incubation at
42.degree. C. in a solution comprising: 50% formamide, 5.times.SSC
(150 mM NaCl, 15 mM trisodium citrate), 50 mM sodium phosphate
(pH7.6), 5.times.Denhardt's solution, 10% dextran sulfate, and 20
microgram/ml denatured, sheared salmon sperm DNA, followed by
washing the filters in 0.1.times.SSC at about 65.degree. C. The
polynucleotides and polypeptides of the present invention may be
employed as research reagents and materials for discovery of
treatments and diagnostics to animal and human disease.
Vectors, Host Cells, Expression
[0088] The present invention also relates to vectors which comprise
a polynucleotide or polynucleotides of the present invention, and
host cells which are genetically engineered with vectors of the
invention and to the production of polypeptides of the invention by
recombinant techniques. Cell-free translation systems can also be
employed to produce such proteins using RNAs derived from the DNA
constructs of the present invention.
[0089] For recombinant production, host cells can be genetically
engineered to incorporate expression systems or portions thereof
for polynucleotides of the present invention. Introduction of
polynucleotides into host cells can be effected by methods
described in many standard laboratory manuals, such as Davis et
al., BASIC METHODS IN MOLECULAR BIOLOGY (1986) and Sambrook et al.,
MOLECULAR CLONING: A LABORATORY MANUAL, 2nd Ed., Cold Spring Harbor
Laboratory Press, Cold Spring Harbor, N.Y. (1989) such as calcium
phosphate transfection, DEAE-dextran mediated transfection,
transvection, microinjection, cationic lipid-mediated transfection,
electroporation, transduction, scrape loading, ballistic
introduction or infection.
[0090] Representative examples of appropriate hosts include
bacterial cells, such as streptococci, staphylococci, E. coli,
Streptomyces and Bacillus subtilis cells; fungal cells, such as
yeast cells and Aspergillus cells; insect cells such as Drosophila
S2 and Spodoptera Sf9 cells; animal cells such as CHO, COS, HeLa,
C127, 3T3, BHK, HEK 293 and Bowes melanoma cells; and plant
cells.
[0091] A great variety of expression systems can be used. Such
systems include, among others, chromosomal, episomal and
virus-derived systems, e.g., vectors derived from bacterial
plasmids, from bacteriophage, from transposons, from yeast
episomes, from insertion elements, from yeast chromosomal elements,
from viruses such as baculoviruses, papova viruses, such as SV40,
vaccinia viruses, adenoviruses, fowl pox viruses, pseudorabies
viruses and retroviruses, and vectors derived from combinations
thereof, such as those derived from plasmid and bacteriophage
genetic elements, such as cosmids and phagemids. The expression
systems may contain control regions that regulate as well as
engender expression. Generally, any system or vector suitable to
maintain, propagate or express polynucleotides to produce a
polypeptide in a host may be used. The appropriate nucleotide
sequence may be inserted into an expression system by any of a
variety of well-known and routine techniques, such as, for example,
those set forth in Sambrook et al., MOLECULAR CLONING, A LABORATORY
MANUAL (supra).
[0092] For secretion of the translated protein into the lumen of
the endoplasmic reticulum, into the periplasmic space or into the
extracellular environment, appropriate secretion signals may be
incorporated into the desired polypeptide. These signals may be
endogenous to the polypeptide or they may be heterologous
signals.
[0093] If the receptor polypeptide is to be expressed for use in
screening assays, generally, it is preferred that the polypeptide
be produced at the surface of the cell. In this event, the cells
may be harvested prior to use in the screening assay. If the
receptor polypeptide is secreted into the medium, the medium can be
recovered in order to recover and purify the polypeptide; if
produced intracellularly, the cells must first be lysed before the
polypeptide is recovered.
[0094] Receptor polypeptides can be recovered and purified from
recombinant cell cultures by well-known methods including ammonium
sulfate or ethanol precipitation, acid extraction, anion or cation
exchange chromatography, phosphocellulose chromatography,
hydrophobic interaction chromatography, affinity chromatography,
hydroxylapatite chromatography and lectin chromatography. Most
preferably, high performance liquid chromatography is employed for
purification. Well known techniques for refolding proteins may be
employed to regenerate active conformation when the polypeptide is
denatured during isolation and or purification.
Diagnostic Assays
[0095] This invention also relates to the use of receptor
polynucleotides or polypeptides for use as diagnostic reagents.
Detection of a mutated form of the receptor gene associated with a
dysfunction will provide a diagnostic tool that can add to or
define a diagnosis of a disease or susceptibility to a disease
which results from under-expression, over-expression or altered
expression of the receptor. Individuals carrying mutations in the
receptor gene may be detected at the DNA level by a variety of
techniques.
[0096] Nucleic acids for diagnosis may be obtained from a subject's
cells, such as from blood, urine, saliva, tissue biopsy or autopsy
material. The genomic DNA may be used directly for detection or may
be amplified enzymatically by using PCR or other amplification
techniques prior to analysis. RNA or cDNA may also be used in
similar fashion. Deletions and insertions can be detected by a
change in size of the amplified product in comparison to the normal
genotype. Point mutations can be identified by hybridizing
amplified DNA to labeled receptor nucleotide sequences. Perfectly
matched sequences can be distinguished from mismatched duplexes by
RNase digestion or by differences in melting temperatures. DNA
sequence differences may also be detected by alterations in
electrophoretic mobility of DNA fragments in gels, with or without
denaturing agents, or by direct DNA sequencing. (See, e.g., Myers
et al., Science (1985) 230:1242.) Sequence changes at specific
locations may also be revealed by nuclease protection assays, such
as RNase and S1 protection or the chemical cleavage method. (See
Cotton et al., Proc Natl Acad Sci USA (1985) 85: 4397-4401.) In
another embodiment, an array of oligonucleotides probes comprising
receptor nucleotide sequence or fragments thereof can be
constructed to conduct efficient screening of e.g., genetic
mutations. Array technology methods are well known and have general
applicability and can be used to address a variety of questions in
molecular genetics including gene expression, genetic linkage, and
genetic variability. (See for example: M. Chee et al., Science, Vol
274, pp 610-613 (1996).)
[0097] The diagnostic assays offer a process for diagnosing or
determining a susceptibility to specific diseases through detection
of mutation in the receptor gene by the methods described.
[0098] In addition, specific diseases can be diagnosed by methods
comprising determining from a sample derived from a subject an
abnormally decreased or increased level of receptor polypeptide or
receptor mRNA. Decreased or increased expression can be measured at
the RNA level using any of the methods well known in the art for
the quantitation of polynucleotides, such as, for example, PCR,
RT-PCR, RNase protection, Northern blotting and other hybridization
methods. Assay techniques that can be used to determine levels of a
protein in a sample derived from a host are well-known to those of
skill in the art. Such assay methods include radioimmunoassays,
competitive-binding assays, Western Blot analysis and ELISA
assays.
[0099] Thus in another aspect, the present invention relates to a
diagnostic kit for a disease or susceptibility to a disease which
comprises:
[0100] (a) a receptor polynucleotide, preferably the nucleotide
sequence of SEQ ID NO:X, or a fragment thereof;
[0101] (b) a nucleotide sequence complementary to that of (a);
[0102] (c) a receptor polypeptide, preferably the polypeptide of
SEQ ID NO:Y, or a fragment thereof; or
[0103] (d) an antibody to a receptor polypeptide, preferably to the
polypeptide of SEQ ID NO: Y.
[0104] It will be appreciated that in any such kit, (a), (b), (c)
or (d) may comprise a substantial component.
Chromosome Assays
[0105] The nucleotide sequences of the present invention are also
valuable for chromosome identification. The sequence is
specifically targeted to and can hybridize with a particular
location on an individual human chromosome. The mapping of relevant
sequences to chromosomes according to the present invention is an
important first step in correlating those sequences with gene
associated disease. Once a sequence has been mapped to a precise
chromosomal location, the physical position of the sequence on the
chromosome can be correlated with genetic map data. Such data are
found, for example, in V. McKusick, Mendelian Inheritance in Man
(available on line through Johns Hopkins University Welch Medical
Library). The relationship between genes and diseases that have
been mapped to the same chromosomal region are then identified
through linkage analysis (coinheritance of physically adjacent
genes).
[0106] The differences in the cDNA or genomic sequence between
affected and unaffected individuals can also be determined. If a
mutation is observed in some or all of the affected individuals but
not in any normal individuals, then the mutation is likely to be
the causative agent of the disease.
Antibodies
[0107] The polypeptides of the invention or their fragments or
analogs thereof, or cells expressing them can also be used as
immunogens to produce antibodies immunospecific for the receptor
polypeptides. The term "immunospecific" means that the antibodies
have substantially greater affinity for the polypeptides of the
invention than their affinity for other related polypeptides in the
prior art.
[0108] Antibodies generated against the receptor polypeptides can
be obtained by administering the polypeptides or epitope-bearing
fragments, analogs or cells to an animal, preferably a nonhuman,
using routine protocols. For preparation of monoclonal antibodies,
any technique which provides antibodies produced by continuous cell
line cultures can be used. Examples include the hybridoma technique
(Kohler, G. and Milstein, C., Nature (1975) 256:495-497), the
trioma technique, the human B-cell hybridoma technique (Kozbor et
al., Immunology Today (1983) 4:72) and the EBV-hybridoma technique
(Cole et al., MONOCLONAL ANTIBODIES AND CANCER THERAPY, pp. 77-96,
Alan R. Liss, Inc., 1985).
[0109] Techniques for the production of single chain antibodies
(U.S. Pat. No. 4,946,778) can also be adapted to produce single
chain antibodies to polypeptides of this invention. Also,
transgenic mice, or other organisms including other mammals, may be
used to express humanized antibodies.
[0110] The above-described antibodies may be employed to isolate or
to identify clones expressing the polypeptide or to purify the
polypeptides by affinity chromatography.
[0111] Antibodies against receptor polypeptides may also be
employed to treat diseases.
Vaccines
[0112] Another aspect of the invention relates to a method for
inducing an immunological response in a mammal which comprises
inoculating the mammal with a receptor polypeptide, or a fragment
thereof, adequate to produce antibody and/or T cell immune response
to protect said animal from a disease. Yet another aspect of the
invention relates to a method of inducing immunological response in
a mammal which comprises, delivering a receptor polypeptide via a
vector directing expression of the receptor polynucleotide in vivo
in order to induce such an immunological response to produce
antibody to protect said animal from diseases.
[0113] Further aspect of the invention relates to an
immunological/vaccine formulation (composition) which, when
introduced into a mammalian host, induces an immunological response
in that mammal to a receptor polypeptide wherein the composition
comprises a receptor polypeptide or receptor gene. The vaccine
formulation may further comprise a suitable carrier. Since a
receptor polypeptide may be broken down in the stomach, it is
preferably administered parenterally (including subcutaneous,
intramuscular, intravenous, intradermal etc. injection).
Formulations suitable for parenteral administration include aqueous
and non-aqueous sterile injection solutions which may contain
anti-oxidants, buffers, bacteriostats and solutes which render the
formulation instonic with the blood of the recipient; and aqueous
and non-aqueous sterile suspensions which may include suspending
agents or thickening agents. The formulations may be presented in
unit-dose or multi-dose containers, for example, sealed ampoules
and vials and may be stored in a freeze-dried condition requiring
only the addition of the sterile liquid carrier immediately prior
to use. The vaccine formulation may also include adjuvant systems
for enhancing the immunogenicity of the formulation, such as oil-in
water systems and other systems known in the art. The dosage will
depend on the specific activity of the vaccine and can be readily
determined by routine experimentation.
Screening Assays
[0114] The receptor polypeptide of the present invention may be
employed in a screening process for compounds which bind the
receptor and which activate (agonists) or inhibit activation of
(antagonists) the receptor polypeptide of the present invention.
Thus, polypeptides of the invention may also be used to assess the
binding of small molecule substrates and ligands in, for example,
cells, cell-free preparations, chemical libraries, and natural
product mixtures. These substrates and ligands may be natural
substrates and ligands or may be structural or functional mimetics.
See Coligan et al., Current Protocols in Immunology 1(2):Chapter 5
(1991).
[0115] The receptor polypeptides are responsible for many
biological functions, including many pathologies. Accordingly, it
is desirous to find compounds and drugs which stimulate the
receptor on the one hand and which can inhibit the function of the
receptor on the other hand. In general, agonists are employed for
therapeutic and prophylactic purposes for such conditions and
diseases. Antagonists may be employed for a variety of therapeutic
and prophylactic purposes for such conditions and diseases.
[0116] In general, such screening procedures involve producing
appropriate cells which express the receptor polypeptide of the
present invention on the surface thereof. Such cells include cells
from mammals, yeast, Drosophila or E. coli. Cells expressing the
receptor (or cell membrane containing the expressed receptor) are
then contacted with a test compound to observe binding, or
stimulation or inhibition of a functional response.
[0117] The assays may simply test binding of a candidate compound
wherein adherence to the cells bearing the receptor is detected by
means of a label directly or indirectly associated with the
candidate compound or in an assay involving competition with a
labeled competitor. Further, these assays may test whether the
candidate compound results in a signal generated by activation of
the receptor, using detection systems appropriate to the cells
bearing the receptor at their surfaces. Inhibitors of activation
are generally assayed in the presence of a known agonist and the
effect on activation by the agonist by the presence of the
candidate compound is observed.
[0118] Further, the assays may simply comprise the steps of mixing
a candidate compound with a solution containing a receptor
polypeptide to form a mixture, measuring receptor activity in the
mixture, and comparing the receptor activity of the mixture to a
standard.
[0119] The receptor cDNA, protein and antibodies to the protein may
also be used to configure assays for detecting the effect of added
compounds on the production of receptor mRNA and protein in cells.
For example, an ELISA may be constructed for measuring secreted or
cell associated levels of receptor protein using monoclonal and
polyclonal antibodies by standard methods known in the art, and
this can be used to discover agents which may inhibit or enhance
the production of the receptor (also called antagonist or agonist,
respectively) from suitably manipulated cells or tissues. Standard
methods for conducting screening assays are well understood in the
art.
[0120] Examples of potential receptor antagonists include
antibodies or, in some cases, oligonucleotides or proteins which
are closely related to the ligand of the receptor, e.g., a fragment
of the ligand, or small molecules which bind to the receptor but do
not elicit a response, so that the activity of the receptor is
prevented.
[0121] Thus in another aspect, the present invention relates to a
screening kit for identifying agonists, antagonists, ligands,
receptors, substrates, enzymes, etc. for receptor polypeptides; or
compounds which decrease or enhance the production of receptor,
which comprises:
[0122] (a) a receptor polypeptide, preferably that of SEQ ID
NO:Y;
[0123] (b) a recombinant cell expressing a receptor polypeptide,
preferably that of SEQ ID NO:Y;
[0124] (c) a cell membrane expressing a receptor polypeptide;
preferably that of SEQ ID NO: Y; or
[0125] (d) antibody to a receptor polypeptide, preferably that of
SEQ ID NO: Y.
[0126] It will be appreciated that in any such kit, (a), (b), (c)
or (d) may comprise a substantial component.
Prophylactic and Therapeutic Methods
[0127] This invention provides methods of treating an abnormal
conditions related to both an excess of and insufficient amounts of
receptor activity.
[0128] If the activity of the receptor is in excess, several
approaches are available. One approach comprises administering to a
subject an inhibitor compound (antagonist) as described along with
a pharmaceutically acceptable carrier in an amount effective to
inhibit activation by blocking the binding of ligands to the
receptor or by inhibiting a second signal, and thereby alleviating
the abnormal condition.
[0129] In another approach, soluble forms of the receptor
polypeptides still capable of binding the ligand in competition
with endogenous receptor may be administered. Typical embodiments
of such competitors comprise fragments of the receptor
polypeptide.
[0130] In still another approach, expression of the gene encoding
endogenous receptor can be inhibited using expression blocking
techniques. Known such techniques involve the use of antisense
sequences, either internally generated or separately administered.
(See, for example, O'Connor, J Neurochem (1991) 56:560 in
Oligodeoxynucleotides as Antisense Inhibitors of Gene Expression,
CRC Press, Boca Raton, Fla. (1988).) Alternatively,
oligonucleotides which form triple helices with the gene can be
supplied. (See, for example, Lee et al., Nucleic Acids Res (1979)
6:3073; Cooney et al., Science (1988) 241:456; Dervan et al.,
Science (1991) 251:1360.) These oligomers can be administered per
se or the relevant oligomers can be expressed in vivo.
[0131] For treating abnormal conditions related to an
under-expression of the receptor and its activity, several
approaches are also available. One approach comprises administering
to a subject a therapeutically effective amount of a compound which
activates the receptor, i.e., an agonist as described above, in
combination with a pharmaceutically acceptable carrier, to thereby
alleviate the abnormal condition. Alternatively, gene therapy may
be employed to effect the endogenous production of the receptor by
the relevant cells in the subject. For example, a polynucleotide of
the invention may be engineered for expression in a replication
defective retroviral vector, as discussed above. The retroviral
expression construct may then be isolated and introduced into a
packaging cell transduced with a retroviral plasmid vector
containing RNA encoding a polypeptide of the present invention such
that the packaging cell now produces infectious viral particles
containing the gene of interest. These producer cells may be
administered to a subject for engineering cells in vivo and
expression of the polypeptide in vivo. For overview of gene
therapy, see Chapter 20, Gene Therapy and other Molecular
Genetic-based Therapeutic Approaches, (and references cited
therein) in Human Molecular Genetics, T Strachan and A P Read, BIOS
Scientific Publishers Ltd (1996).
Formulation and Administration
[0132] Peptides, such as the soluble form of receptor polypeptides,
and agonists and antagonist peptides or small molecules, may be
formulated in combination with a suitable pharmaceutical carrier.
Such formulations comprise a therapeutically effective amount of
the polypeptide or compound, and a pharmaceutically acceptable
carrier or excipient. Such carriers include but are not limited to,
saline, buffered saline, dextrose, water, glycerol, ethanol, and
combinations thereof. Formulation should suit the mode of
administration, and is well within the skill of the art. The
invention further relates to pharmaceutical packs and kits
comprising one or more containers filled with one or more of the
ingredients of the aforementioned compositions of the
invention.
[0133] Polypeptides and other compounds of the present invention
may be employed alone or in conjunction with other compounds, such
as therapeutic compounds. Preferred forms of systemic
administration of the pharmaceutical compositions include
injection, typically by intravenous injection. Other injection
routes, such as subcutaneous, intramuscular, or intraperitoneal,
can be used. Alternative means for systemic administration include
transmucosal and transdermal administration using penetrants such
as bile salts or fusidic acids or other detergents. In addition, if
properly formulated in enteric or encapsulated formulations, oral
administration may also be possible. Administration of these
compounds may also be topical and/or localized, in the form of
salves, pastes, gels and the like.
[0134] The dosage range required depends on the choice of peptide,
the route of administration, the nature of the formulation, the
nature of the subject's condition, and the judgment of the
attending practitioner. Suitable dosages, however, are in the range
of 0.1-100 .mu.g/kg of subject. Wide variations in the needed
dosage, however, are to be expected in view of the variety of
compounds available and the differing efficiencies of various
routes of administration. For example, oral administration would be
expected to require higher dosages than administration by
intravenous injection. Variations in these dosage levels can be
adjusted using standard empirical routines for optimization, as is
well understood in the art.
[0135] Polypeptides used in treatment can also be generated
endogenously in the subject, in treatment modalities often referred
to as "gene therapy" as described above. Thus, for example, cells
from a subject may be engineered with a polynucleotide, such as a
DNA or RNA, to encode a polypeptide ex vivo, and for example, by
the use of a retroviral plasmid vector. The cells are then
introduced into the subject.
[0136] All publications, including but not limited to patents and
patent applications, cited in this specification are herein
incorporated by reference as if each individual publication were
specifically and individually indicated to be incorporated by
reference herein as though fully set forth. ##STR1## ##STR2##
##STR3## ##STR4## ##STR5## ##STR6## ##STR7## ##STR8## ##STR9##
##STR10## ##STR11## ##STR12## ##STR13## ##STR14## ##STR15##
##STR16## ##STR17## ##STR18## ##STR19## ##STR20## ##STR21##
##STR22## ##STR23## ##STR24## ##STR25## ##STR26##
Sequence CWU 1
1
63 1 1497 DNA Homo sapiens 1 gcagttcctg agagaagaac cctgaggaac
agacgttccc tcgcggccct ggcacctcca 60 accccagata tgctgctgct
gctgctgctg cccctgctct gggggaggga gagggtggaa 120 ggacagaaga
gtaaccggaa ggattactcg ctgacgatgc agagttccgt gaccgtgcaa 180
gagggcatgt gtgtccatgt gcgctgctcc ttctcctacc cagtggacag ccagactgac
240 tctgacccag ttcatggcta ctggttccgg gcagggaatg atataagctg
gaaggctcca 300 gtggccacaa acaacccagc ttgggcagtg caggaggaaa
ctcgggaccg attccacctc 360 cttggggacc cacagaccaa aaattgcacc
ctgagcatca gagatgccag aatgagtgat 420 gcggggagat acttctttcg
tatggagaaa ggaaatataa aatggaatta taaatatgac 480 cagctctctg
tgaacgtgac ataccctcct cagaacttga ctgtgactgt cttccaagga 540
gaaggcacag catccacagc tctggggaac agctcatctc tttcagtcct agagggccag
600 tctctgcgct tggtctgtgc tgttgacagc aatccccctg ccaggctgag
ctggacctgg 660 aggagtctga ccctgtaccc ctcacagccc tcaaaccctc
tggtactgga gctgcaagtg 720 cacctggggg atgaagggga attcacctgt
cgagctcaga actctctggg ttcccagcac 780 gtttccctga acctctccct
gcaacaggag tacacaggca aaatgaagcc tgtatcagga 840 gtgttgctgg
gggcggtcgg gggaactgga gccacagccc tggtcttcct ctccttctgt 900
gtcatcttca ttgtagtgag gtcctgcagg aagaaatcgg caagaccagc agcggacgtg
960 ggagacatag gcatgaagga tgcaaacacc attcaggggc tcagcctctc
agggtaactg 1020 gatgagtcct gggcagatga taacccccga caccatggcc
tggctgccca ctccctcagg 1080 ggaggaaaga gagatcccag tatgcacccc
tcagctttca taagggggag cctcaggacc 1140 tatccaggtc aagaagccac
caacaatgag tactcagaga tcaagatccc caagtaagaa 1200 aatgcagagg
ctcgggcttg tttgagggtt cacgacccct ccagcaaagg agtctgaggc 1260
tgattccagt agaattagca gccctcaatg ctgtgcaaca agacatcaga acttattcct
1320 cttgtctaac tgaaaatgca tgcctgatga ccaaactctc cctttcccca
tccaatcggt 1380 ccacactccc cgccctggcc tctgtaccca ccattctcct
ctgtacttct ctaaggatga 1440 ctactttaga ttccgaatat agtgagattg
taacgtgaaa aaaaaaaaaa aaaaaaa 1497 2 1849 DNA Homo sapiens 2
cggcacgagt ggacaaccat cagggagcca ggacacagag gggcagagca agtcagcatt
60 ggcgcccctt cctcagatcc ctatcatctt gggaaacagt agcccagagg
ttcaggaaga 120 tgttaactta aatgttcggg gtgccccagt ctgttcagca
tggctgaaat ccacactccg 180 tattcttcct tgaagaaact gttatcttta
ctcaatggct tcgtggctgt gtctggcatc 240 atcctagttg gcctgggcat
tggtggtaaa tgtggagggg cctctctgac gaatgtcctc 300 gggctgtcct
ccgcatacct ccttcacgtt ggcaacctgt gcctggtgat gggatgcatc 360
acggtactgc ttggctgtgc cgggtggtat ggagcgacta aagagagcag aggcacgctc
420 ttgttttgca tcctgtcaat ggttattgtc ctcatcatgg aagttacagc
tgccacagtg 480 gtccttcttt tctttccaat tgttggagat gtggccttgg
aacacacctt cgtgaccctg 540 aggaagaatt acagaggtta caacgagcca
gacgactatt ctacacagtg gaacttggtc 600 atggagaagc taaagtgctg
tggggtgaat aactacacag atttttctgg ctcttccttc 660 gaaatgacaa
cgggccacac ctaccccagg agttgctgta aatccatcgg aagtgtgtcc 720
tgtgacggac gcgatgtgtc tccaaacgtc atccaccaga agggctgttt ccataaactc
780 ctaaaaatca ccaagactca gagcttcacc ctgagtggga gctctctggg
agctgcagtg 840 atacagaggt gggggtctcg ctatgttgcg caggctggtc
ttgaactgct ggcctaaagc 900 gatccccccg cctaggcctc ccaaagtgct
gggtttacca gcgtgagcca ccacgctggg 960 cttcctgcat ccttttaagg
ttcctgaggg tctgcctgag aggagctgtc cctgaatctc 1020 catgcagccc
cacctgccac atcaccaaga catacaatct ttgccagcaa cacttcctcc 1080
ttgcagatta caagcatagc taatgccacc accagacaag accgattcgc tggcctccat
1140 ttcttcaacc cagtgcctgt catgaaactt gtggaggtca ttaaaacacc
aatgaccagc 1200 cagaagacat ttgaatcttt ggtagacttt agcaaaaccc
taggaaagca tcctgtttct 1260 tgcaaggaca ctcctgggtt tattgtgaac
cgcctcctgg ttccatacct catggaagca 1320 atcaggctgt atgaacgagg
gcctcctggc tttccctgtg ggcttctgag aaaggtttct 1380 ggaactccca
ccacccccac tacagtccca gccagagcaa ttgcatggcc ggcccagatt 1440
gatatcctgg atctctgctt ttgattaaaa ggtgacgcat ccaaagaaga cattgacact
1500 gctatgaaat taggagccgg ttaccccatg ggcccatttg agcttctaga
ttatgtcgga 1560 ctggatacta cgaagttcat cgtggatggg tggcatgaaa
tggatgcaga gaacccatta 1620 catcagccca gcccatcctt aaataagctg
gtagcagaga acaagttcgg caagaagact 1680 ggagaaggat tttacaaata
caagtgatgt gcagcttctc cggttctgag aagaacacct 1740 gagagcgctt
tccagccagt gccccgagtg cctgtgggaa tgctctttgg tcagacattc 1800
cctcacacag tacagtttaa taaatgtgca ttttgattgt aaaaaaaaa 1849 3 741
DNA Homo sapiens 3 atggctgaaa tccacactcc gtattcttcc ttgaagaaac
tgttatcttt actcaatggc 60 ttcgtggctg tgtctggcat catcctagtt
ggcctgggca ttggtggtaa atgtggaggg 120 gcctctctga cgaatgtcct
cgggctgtcc tccgcatacc tccttcacgt tggcaacctg 180 tgcctggtga
tgggatgcat cacggtactg cttggctgtg ccgggtggta tggagcgact 240
aaagagagca gaggcacgct cttgttttgc atcctgtcaa tggttattgt cctcatcatg
300 gaagttacag ctgccacagt ggtccttctt ttctttccaa ttgttggaga
tgtggccttg 360 gaacacacct tcgtgaccct gaggaagaat tacagaggtt
acaacgagcc agacgactat 420 tctacacagt ggaacttggt catggagaag
ctaaagtgct gtggggtgaa taactacaca 480 gatttttctg gctcttcctt
cgaaatgaca acgggccaca cctaccccag gagttgctgt 540 aaatccatcg
gaagtgtgtc ctgtgacgga cgcgatgtgt ctccaaacgt catccaccag 600
aagggctgtt tccataaact cctaaaaatc accaagactc agagcttcac cctgagtggg
660 agctctctgg gagctgcagt gatacagagg tgggggtctc gctatgttgc
gcaggctggt 720 cttgaactgc tggcctaaag c 741 4 1288 DNA Homo sapiens
4 ggcgtccctc tgcctgccca ctcagtggca acacccggga gctgttttgt cctttgtgga
60 gcctcagcag ttccctcttt cagaactcac tgccaagagc cctgaacagg
agccaccatg 120 cagtgcttca gcttcattaa gaccatgatg atcctcttca
atttgctcat ctttctgtgt 180 ggtgcagccc tgttggcagt gggcatctgg
gtgtcaatcg atggggcatc ctttctgaag 240 atcttcgggc cactgtcgtc
cagtgccatg cagtttgtca acgtgggcta cttcctcatc 300 gcagccggcg
ttgtggtctt tgctcttggt ttcctgggct gctatggtgc taagactgag 360
agcaagtgtg ccctcgtgac gttcttcttc atcctcctcc tcatcttcat tgctgaggtt
420 gcagctgctg tggtcgcctt ggtgtacacc acaatggctg agcacttcct
gacgttgctg 480 gtagtgcctg ccatcaagaa agattatggt tcccaggaag
acttcactca agtgtggaac 540 accaccatga aagggctcaa gtgctgtggc
ttcaccaact atacggattt tgaggactca 600 ccctacttca aagagaacag
tgcctttccc ccattctgtt gcaatgacaa cgtcaccaac 660 acagccaatg
aaacctgcac caagcaaaag gctcacgacc aaaaagtaga gggttgcttc 720
aatcagcttt tgtatgacat ccgaactaat gcagtcaccg tgggtggtgt ggcagctgga
780 attgggggcc tcgagctggc tgccatgaat tgtgtccatg tatctgtact
gcaatctaca 840 ataagtccac ttctgcctct gccactactg ctgccacatg
ggaactgtga agaggcaccc 900 tggcaagcag cagtgattgg gggaggggac
aggatctaac aatgtcactt gggccagaat 960 ggacctgccc tttctgctcc
agacttgggg ctagataggg accactcctt ttaggcgatg 1020 cctgactttc
cttccattgg tgggtggatg ggtggggggc attccagagc ctctaaggta 1080
gccagttctg ttgcccattc ccccagtcta ttaaaccctt gatatgcccc ctaggcctag
1140 tggtgatccc agtgctctac tgggggatga gagaaaggca ttttatagcc
tgggcataag 1200 tgaaatcagc agagcctctg ggtggatgtg tagaaggcac
ttcaaaatgc ataaacctgt 1260 tacaatgtta aaaaaaaaaa aaaaaaaa 1288 5
1236 DNA Homo sapiens 5 aaaaaaaaca aggtccccac agcaaagaaa aggaatagga
tcaagagata cgtggctgct 60 ggcagagcaa gcatgaattc gatgacttca
gcagttccgg tggccaattc tgtgttggtg 120 gtggcacccc acaatggtta
tcctgtgacc ccaggaatta tgtctcacgt gcccctgtat 180 ccaaacagcc
agccgcaagt ccacctagtt cctgggaacc cacctagttt ggtgtcgaat 240
gtgaatgggc agcctgtgca gaaagctctg aaagaaggca aaaccttggg ggccatccag
300 atcatcattg gcctggctca catcggcctc ggctccatca tggcgacggt
tctcgtaggg 360 gaatacctgt ctatttcatt ctacggaggc tttcccttct
ggggaggctt gtggtttatc 420 atttcaggat ctctctccgt ggcagcagaa
aatcagccat attcttattg cctgctgtct 480 ggcagtttgg gcttgaacat
cgtcagtgca atctgctctg cagttggagt catactcttc 540 atcacagatc
taagtattcc ccacccatat gcctaccccg actattatcc ttacgcctgg 600
ggtgtgaacc ctggaatggc gatttctggc gtgctgctgg tcttctgcct cctggagttt
660 ggcatcgcat gcgcatcttc ccactttggc tgccagttgg tctgctgtca
atcaagcaat 720 gtgagtgtca tctatccaaa catctatgca gcaaacccag
tgatcacccc agaaccggtg 780 acctcaccac caagttattc cagtgagatc
caagcaaata agtaaggcta cagattctgg 840 aagcatcttt cactgggacc
aaaagaagtc ctcctccctt tctgggcttc cataacccag 900 gtcgttcctg
ttctgacagc tgaggaaacg tctctcccac tgtttgtact ctcaccttca 960
ttcttcaatt cagtctagga aaccatgctg tttctctatc aagaagaaga cagagatttt
1020 aaacagatgt taaccaagag ggactcccta gggcacatgc atcagcacat
atgtgggcat 1080 ccagcctctg gggccttggc acacccattc gtgtgctctg
ctgcatgtga gcttgtgggt 1140 tagaggaaca aatatctaga cattcaatct
tcactctttc aattgtgcat tcatttaata 1200 aatagatact gagcattcaa
aaaaaaaaaa aaaaaa 1236 6 1115 DNA Homo sapiens 6 cacgagcagg
gtctcgggct agtcatggcg tccccgtctc ggagactgca gactaaacca 60
gtcattactt gtttcaagag cgttctgcta atctacactt ttattttctg gatcactggc
120 gttatccttc ttgcagttgg catttggggc aaggtgagcc tggagaatta
cttttctctt 180 ttaaatgaga aggccaccaa tgtccccttc gtgctcattg
ctactggtac cgtcattatt 240 cttttgggca cctttggttg ttttgctacc
tgccgagctt ctgcatggat gctaaaactg 300 tatgcaatgt ttctgactct
cgtttttttg gtcgaactgg tcgctgccat cgtaggattt 360 gttttcagac
atgagattaa gaacagcttt aagaataatt atgagaaggc tttgaagcag 420
tataactcta caggagatta tagaagccat gcagtagaca agatccaaaa tacgttgcat
480 tgttgtggtg tcaccgatta tagagattgg acagatacta attattactc
agaaaaagga 540 tttcctaaga gttgctgtaa acttgaagat tgtactccac
agagagatgc agacaaagta 600 aacaatgaag gttgttttat aaaggtgatg
accattatag agtcagaaat gggagtcgtt 660 gcaggaattt cctttggagt
tgcttgcttc caactgattg gaatctttct cgcctactgc 720 ctctctcgtg
ccataacaaa taaccagtat gagatagtgt aacccaatgt atctgtgggc 780
ctattcctct ctacctttaa ggacatttag ggtcccccct gtgaattaga aagttgcttg
840 gctggagaac tgacaacact acttactgat agaccaaaaa actacaccag
taggttgatt 900 caatcaagat gtatgtagac ctaaaactac accaataggc
tgattcaatc aagatccgtg 960 ctcgcagtgg gctgattcaa tcaagatgta
tgtttgctat gttctaagtc caccttctat 1020 cccattcatg ttagatcgtt
gaaaccctgt atccctctga aacactggaa gagctagtaa 1080 attgtaaatg
aagtaaaaaa aaaaaaaaaa aaaaa 1115 7 1662 DNA Homo sapiens 7
cacgagcatt gccgctctct cggtgagcgc agccccgctc tccgggccgg gccttcgcgg
60 gccaccggcg ccatgggcca gtgcggcatc acctcctcca agaccgtgct
ggtctttctc 120 aacctcatct tctggggggc agctggcatt ttatgctatg
tgggagccta tgtcttcatc 180 acttatgatg actatgacca cttctttgaa
gatgtgtaca cgctcatccc tgctgtagtg 240 atcatagctg taggagccct
gcttttcatc attgggctaa ttggctgctg tgccacaatc 300 cgggaaagtc
gctgtggact tgccacgttt gtcatcatcc tgctcttggt ttttgtcaca 360
gaagttgttg tagtggtttt gggatatgtt tacagagcaa aggtggaaaa tgaggttgat
420 cgcagcattc agaaagtgta taagacctac aatggaacca accctgatgc
tgctagccgg 480 gctattgatt atgtacagag acagctgcat tgttgtggaa
ttcacaacta ctcagactgg 540 gaaaatacag attggttcaa agaaaccaaa
aaccagagtg tccctcttag ctgctgcaga 600 gagactgcca gcaattgtaa
tggcagcctg gcccaccctt ccgacctcta tgctgagggg 660 tgtgaggctc
tagttgtgaa gaagctacaa gaaatcatga tgcatgtgat ctgggccgca 720
ctggcatttg cagctattca gctgctgggc atgctgtgtg cttgcatcgt gttgtgcaga
780 aggagtagag atcctgctta cgagctcttc atcactggcg gaacctatgc
atagttgaca 840 atctcaagcc tgagcttttt ggtcttgttc tgatttggaa
ggtgaattga gcaggtctgc 900 tgctgttggc ctctggagtt catttagtta
aagcacatgt acactggtgt tggacagagc 960 agcttggctt ttcatgtgcc
cacctactta cctactacct gcgactttct ttttccttgt 1020 tctagctgac
tcttcatgcc cctaagattt taagtacgat ggtgaacgtt ctaatttcag 1080
aaccaattgc gagtcatgta gtgtggtaga attaaaggag gacacgagcc tgcttctgtt
1140 acctccaagt ggtaacagga ctgatgccga aatgtcacca ggtcctttca
gtcttcacag 1200 tggagaactc ttggccaaag gtttttgggg ggaggaggag
gaaaccagct ttctggttaa 1260 ggttaacacc agatggtgcc cctcattggt
gtccttttaa aaaatattta ctgtagtcca 1320 ataagatagc agctgtacaa
aatgactaaa atagattgta ggatcatatg gcgtatatct 1380 tggttcatct
tcaaaatcag agactgagct ttgaaactag tggtttttaa tcaaagttgg 1440
ctttatagga ggagtataat gtatgcacta ctgttttaaa agaattagtg tgagtgtgtt
1500 tttgtatgaa tgagcccatt catggtaagt cttaagcttg ttggaaataa
tgtacccatg 1560 tagactagca aaatagtatg tagatgtgat ctcagttgta
aatagaaaaa tctaattcaa 1620 taaactctgt atcagccccc aacaaaaaaa
aaaaaaaaaa aa 1662 8 1345 DNA Homo sapiens 8 cacgagcgca gagcttgggg
cttccttggt cgcacccacc acctgcctgc ccactggtca 60 gccttcaggg
accctgagca ccgcctggtc tctttcctgt ggccagccca gaactgaagc 120
gctgcggcat ggcgcgcgcc tgcctccagg ccgtcaagta cctcatgttc gccttcaacc
180 tgttcttctg gctgggaggc tgtggcgtgc tgggtgtcgg catctggctg
gccgccacac 240 aggggagctt cgccacgctg tcttcttcct tcccgtccct
gtcggctgcc aacctgctca 300 tcatcaccgg cgcctttgtc atggccatcg
gcttcgtggg ctgcctgggt gccatcaagg 360 agaacaagtg cctcctgctc
actttcttcc tgctgctgct gctggtgttc ctgctggagg 420 ccaccatcgc
catcctcttc ttcgcctaca cggacaagat tgacaggtat gcccagcaag 480
acctgaagaa aggcttgcac ctgtacggca cgcagggcaa cgtgggcctc accaacgcct
540 ggagcatcat ccagaccgac ttccgctgct gtggcgtctc caactacact
gactggttcg 600 aggtgtacaa cgccacgcgg gtacctgact cctgctgctt
ggagttcagt gagagctgtg 660 ggctgcacgc ccccggcacc tggtggaagg
cgccgtgcta cgagacggtg aaggtgtggc 720 ttcaggagaa cctgctggct
gtgggcatct ttgggctgtg cacggcgctg gtgcagatcc 780 tgggcctgac
cttcgccatg accatgtact gccaagtggt caaggcagac acctactgcg 840
cgtaggccgc ccaccgccgg cttctctgcc aaaaggacgc ccacggggag atggccgcac
900 ccacagctgc ttttcccacc accagcttcg gtgttctgcc ccatgctggg
aggagggagg 960 gagggacagg tgcctggagc ccccggaacc ctgtttctgg
aaggccctag ctcaggtggc 1020 ttcagggcct ccggaccccc cctgggaggg
gtggccacgt gctggctgcg gaacccaggg 1080 caggggtggg aggggcctcc
agcacttttt atatttacgt attctccaaa gcagtgttca 1140 cacgggagcc
agcctgtggc ccccagcttc ctggaaaaca ggttggcgct ggaggagccg 1200
ggtcttggca tcctggaggt ggccccactg gtcctggtgc tccaggcggg gccgtggacc
1260 cctcacctac attccatagt gggcccgtgg ggctcctggt gcatcttaat
aaagtgtgag 1320 cagcaaaaaa aaaaaaaaaa aaaaa 1345 9 734 DNA Homo
sapiens 9 gcgccgccgg gccgcagcat ggggcgcttc cgcgggggcc tgcggtgcat
caagtacctg 60 ctgcttggct tcaacctgct cttctggctg gctggatcgg
ccgtcattgc ttttggacta 120 tggtttcggt tcggaggtgc cataaaggag
ttatcatcag aggacaagtc cccagagtat 180 ttctatgtgg ggctgtatgt
tctggttgga gccggggccc tgatgatggc cgtggggttc 240 ttcggatgct
gcggagccat gcgggagtcg caatgtgtgc ttggatcatt ttttacctgc 300
ctcctggtga tatttgctgc tgaagtaacc actggagtat ttgcttttat aggcaagggg
360 gtagctatcc gacatgttca gaccatgtat gaagaggctt acaatgatta
ccttaaagac 420 aggggaaaag gcaatgggac actcatcacc ttccactcaa
catttcagtg ctgtggaaaa 480 gaaagctccg aacaggtcca acctacatgc
ccaaaggagc ttctaggaca caagaattgc 540 atcgatgaaa ttgagaccat
aatcagtgtt aagctccagc tcattggaat tgtcggtatt 600 ggaattgcag
gtctgacgat ctttggcatg atattcagca tggtcctctg ctgtgcgata 660
cgaaactcac gagatgtgat atgaagctac ttctacatga aaattgcaat ctaaagcttt
720 cataccaaat gttc 734 10 577 DNA Homo sapiens 10 agtgtttatg
ggactaaaaa acttttaaca cctttttagg ggaaatattt tggtcctata 60
caaaacatgt aaatatgctt tattactttc attttctgac cctgctgtaa actactgcaa
120 ccctcacatc cctcaaaggg acttttatgt caaactcttc tgtttctcca
aatataagga 180 aaaaagacta aagcaagaga tctggcagtt gaaaattgtg
ggaaagagaa tttgtatggg 240 cactgtatct atgaaatacc tcatacttac
gtttacatgt tttcctaact ttttgtattt 300 ttcttgtata gccacctaga
gaattcttca tagattaaga actacagttt tcaccactta 360 acataagtaa
aacaaagtcc ttcataattt aaccattagc atctttggcc aaaccaaaat 420
aaagaaaagc atcttctcct agttgtgtgt gggcaacaga aacaagttaa ggaaacaaaa
480 atacttatat atacacagaa caaaaataat gttcttttta tgcaaatccc
ctgtgaaaat 540 aaaattttca atgtttaaaa aaaaaaaaaa aaaaaaa 577 11 936
DNA Homo sapiens 11 ttcggcacga gctgcgggcg gtgggcggct gggcggcccc
gggagccgcg ctctcagtct 60 ctctaggcgc agtcccttcg ccgcttccgg
agcccctggc agggcccaga agccatggcc 120 cactataaga ctgagcagga
cgactggctg atcatctact tgaagtattt actctttgtc 180 ttcaacttct
tcttctgggt cgggggagca gccgtcctgg ctgtgggcat ctggaccctg 240
gtggagaaga gtggctacct cagcgtcctg gcctccagca cctttgccgc ctccgcctac
300 atcctcatct ttgcgggcgt acttgtcatg gtgaccggct tcctgggctt
cggtgccatc 360 ctctgggagc ggaagggctg cctctccacg tatttctgcc
tgttgctcgt catcttcctg 420 gttgagctgg tggcgggagt cctggcccat
gtgtattacc agaggctgag tgatgaactg 480 aagcagcact tgaaccggac
tctggctgag aactacgggc agccggagca cgcagatcac 540 gcctcagtgg
accgactcca gcaggatttc aagtgctgcg gaagcaacag ctcagccgac 600
tggcagcaca gcacgtacat cctgttgcgg gaggccgagg gccgccaggt gcccgacagc
660 tgctgcaaga cagtggtggc gcgctgcggc cagcgggccc acccctccaa
catctataag 720 gtggagggag gctgcctcac caagctggag cagttcctgg
ccgaccacct gctgcttatg 780 ggggcagtgg gcatcggggt ggcctgcctg
cagatctgcg ggatggttct cacctgctgc 840 ttgcaccaga ggctccagcg
gcatttttac taatggcaac cacctcctct tccaactgcc 900 cctcaagaca
acatgtggca catgccatct gcaagg 936 12 738 DNA Homo sapiens 12
agcttacttt cactcaccgc ctgtccttcc tgacacctca ccatgtgtac gggaaaatgt
60 gcccgctgtg tggggctctc cctcattacc ctctgcctcg tctgcattgt
ggccaacgcc 120 ctcctgctgg tacctaatgg ggagacctcc tggaccaaca
ccaaccatct cagcttgcaa 180 gtctggctca tgggcggctt cattggcggg
ggcctaatgg tactgtgtcc agggattgca 240 gccgttcggg cagggggcaa
gggctgctgt ggtgctgggt gctgtggaaa ccgctgcagg 300 atgctgcgct
cggtcttctc ctcggcgttc ggggtgcttg gtgccatcta ctgcctctcg 360
gtgtctggag ctgggctccg aaatggaccc agatgcttaa tgaacggcga gtggggctac
420 cacttcgaag acaccgcggg agcttacttg ctcaaccgca ctctatggga
tcggtgcgag 480 gcgccccctc gcgtggtccc ctggaatgtg acgctcttct
cgctgctggt ggccgcctcc 540 tgcctggaga tagtactgtg tgggatccag
ctggtgaacg cgaccattgg tgtcttctgc 600 ggcgattgca ggaaaaaaca
ggacacacct cactgaggct ccactgaccg ccgggttaca 660 cctgctcctt
cctggacgct cactcccttg ctcgctagaa taaactgctt tgcgctctca 720
aaaaaaaaaa aaaaaaac 738 13 1071 DNA Homo sapiens 13 ggcacgagag
attgtcggct gcgggtatat tccaattccc cgtctcctca tgaatatgaa 60
gtgaagggct ctgaccctgg aagtggttct aagcagggca aaatggggtc tcggaagtgt
120 ggaggctgcc taagttgttt gctgattccg cttgcacttt ggagtataat
cgtgaacata 180 ttattgtatt tcccgaatgg gcaaacttcc tatgcatcca
gcaataaact caccaactac 240 gtgtggtatt ttgaaggaat ctgtttctca
ggcatcatga tgcttatagt aacaacagtt 300 cttctggtac tggagaataa
taacaactat aaatgttgcc agagtgaaaa ctgcagcaaa 360 aaatatgtga
cactgctgtc aattatcttt tcttccctcg gaattgcttt ttctggatac 420
tgcctggtca tctctgcctt gggtcttgtc caagggccat attgccgcac ccttgatggc
480 tgggagtatg cttttgaagg cactgctgga cgtttcctta cagattctag
catatggatt 540 cagtgcctgg aacctgcaca tgttgtggag tggaacatca
ttttattttc cattctcata 600 accctcagtg ggcttcaagt gatcatctgc
ctcatcagag tagtcatgca actatccaag 660 atactgtgtg
gaagctattc agtgatcttc cagcctggaa tcatttgaat aaggacaaaa 720
tgttttccat tatcaagaca tggccatcta tctaaatatt atatcaactg tgttagactt
780 gagggcaata ttgaaaatga tggtgctttc tgcatttggt gtttatttgt
aaaaaatttg 840 cagtcctcac tgcacatgca agtataccac ccttccattt
agtatgtttt ttaagtaata 900 tgcatcagaa acttcagaaa tacttctgcc
ctttgatcaa acaaatccat ttccaagaat 960 ctgtactagg gaagtaaata
agaatatgag agaaaccttt atgcaatatg tatattgcaa 1020 cattatttaa
tattctggaa aattggaaac accccaaaat tctaactcaa a 1071 14 865 DNA Homo
sapiens 14 tttgtggagg gcagcagaga gtacccagct ggacatcctt tcctgctgat
gagccccagg 60 ctggaggtgc cctgctcaca tgctcttccc cagggtctct
cgcctgggca ggtcatcata 120 gtacggggac tggtcttgca agagccgaag
cattttactg tgagcctgag ggaccaggct 180 gcccatgctc ctgtgacact
cagggcctcc ttcgcagaca gaactctggc ctggatctcc 240 cgctgggggc
agaagaaact gatctcagcc cccttcctct tttaccccca gagattcttt 300
gaggtgctgc tcctgttcca ggagggaggg ctgaagctgg cgctcaatgg gcaggggctg
360 ggggccacca gcatgaacca gcaggccctg gagcagctgc gggagctccg
gatcagtgga 420 agtgtccagc tctactgtgt ccactcctga aggatggttc
caggaaatac cgcagaaaac 480 aagagtcagc cactccccag ggccccactc
tcctcccctc attaaaccat ccacctgaac 540 accagcacat cagggcctgg
ttcacctctg gggtcacgag actgagtcta caggagcttt 600 gggcctgagg
gaaggcacaa gagtgcaaag gttcctcgaa ctctgcacct tcctccacca 660
ggagcctggg atatggctcc atctgccttc agggcctgga ctgcactcac agaggcaagt
720 gttgtagact aacaaagata ctccaaaata caatggctta aagaatgtgg
tcatttattc 780 tttattattt atttatttgt ggtcaaataa ataaataagg
ttatttattt aaaaaaaaaa 840 aaaaaaaaaa aaaaaaaaaa aaaaa 865 15 441
DNA Homo sapiens 15 gcacgagaga cgacatcaga gatgaggaca gcattgctgc
tccttgcagc cctggctgtg 60 gctacagggc cagcccttac cctgcgctgc
cacgtgtgca ccagctccag caactgcaag 120 cattctgtgg tctgcccggc
cagctctcgc ttctgcaaga ccacgaacac agtggagcct 180 ctgagggctt
ccccgaaagt ctgggaccag gtccaggtgg gcatggaatg ctgatgactt 240
ggagcaggcc ccacagaccc cacagaggat gaagccaccc cacagaggat gcagccccca
300 gctgcatgga aggtggagga cagaagccct gtggatcccc ggatttcaca
ctccttctgt 360 tttgttgccg tttatttttg tactcaaatc tctacatgga
gataaatgat ttaaaccagt 420 aaaaaaaaaa aaaaaaaaaa a 441 16 1066 DNA
Homo sapiens 16 agcgggcccg aaccctcgtg tgaagggtgc agtacctaag
ccggagcggg gtagaggcgg 60 gccggcaccc ccttctgacc tccagtgccg
ccggcctcaa gatcagacat ggcccagaac 120 ttgaaggact tggcgggacg
gctgcccgcc gggccccggg gcatgggcac ggccctgaag 180 ctgttgctgg
gggccggcgc cgtggcctac ggtgtgcgcg aatctgtgtt caccgtggaa 240
ggcgggcaca gagccatctt cttcaatcgg atcggtggag tgcagcagga cactatcctg
300 gccgagggcc ttcacttcag gatcccttgg ttccagtacc ccattatcta
tgacattcgg 360 gccagacctc gaaaaatctc ctcccctaca ggctccaaag
acctacagat ggtgaatatc 420 tccctgcgag tgttgtctcg acccaatgct
caggagcttc ctagcatgta ccagcgccta 480 gggctggact acgaggaacg
agtgttgccg tccattgtca acgaggtgct caagagtgtg 540 gtggccaagt
tcaatgcctc acagctgatc acccagcggg cccaggtatc cctgttgatc 600
cgccgggagc tgacagagag ggccaaggac ttcagcctca tcctggatga tgtggccatc
660 acagagctga gctttagccg agagtacaca gctgctgtag aagccaaaca
agtggcccag 720 caggaggccc agcgggccca attcttggta gaaaaagcaa
agcaggaaca gcggcagaaa 780 attgtgcagg ccgagggtga ggccgaggct
gccaagatgc ttggagaagc actgagcaag 840 aaccctggct acatcaaact
tcgcaagatt cgagcagccc agaatatctc caagacgatc 900 gccacatcac
agaatcgtat ctatctcaca gctgacaacc ttgtgctgaa cctacaggat 960
gaaagtttca ccaggggaag tgacagcctc atcaagggta agaaatgagc ctagtcacca
1020 agaactccac ccccacaaga agtggatctg cttctccagt ttttga 1066 17 704
DNA Homo sapiens 17 ggcacgagat gacatcacta agtggccgat ctgcacagag
caggccagga gcaaccacac 60 aggcttcctg cacatggact gcgagatcaa
gggccgcccc tgctgcatcg gcaccaaggg 120 cagctgtgag atcaccaccc
gggaatactg tgagttcatg cacggctatt tccatgagga 180 agcaacactc
tgctcccagg tgaggcgagg caggcctgga gtagtggagg agaggacgct 240
gggcatggca gcctgctggg gccggggctc acgcactccc tcccatgtcg gagcctcaga
300 ctcaggctgc ttctggggcg ctgagcacca tatgcccatt cccaggtgca
ctgttttgga 360 caaggtgtgt tgggctgctg ccttcctcaa ccctgaggtc
ccagatcagt tttacaggtc 420 tggctgtctc ttttcctaca tgttgggtaa
gaggtcctca atgcccccga acccgacccc 480 tgtgatggac acccaggcgg
acccctgggg aaaggttcct gggccagggt atggtcggtc 540 caacctgccg
aagactactg ctcctgaagt gtctggatga aggccgctgc ctggtgtgtc 600
cctcccccag tgtgggtgca ctgccctcgg tgtcttgtgg gtcttttcaa atgacatccc
660 tgaaggggac ctggaggaag gtggtccggc tggcacccta tcgc 704 18 315 PRT
Homo sapiens 18 Met Leu Leu Leu Leu Leu Leu Pro Leu Leu Trp Gly Arg
Glu Arg Val 1 5 10 15 Glu Gly Gln Lys Ser Asn Arg Lys Asp Tyr Ser
Leu Thr Met Gln Ser 20 25 30 Ser Val Thr Val Gln Glu Gly Met Cys
Val His Val Arg Cys Ser Phe 35 40 45 Ser Tyr Pro Val Asp Ser Gln
Thr Asp Ser Asp Pro Val His Gly Tyr 50 55 60 Trp Phe Arg Ala Gly
Asn Asp Ile Ser Trp Lys Ala Pro Val Ala Thr 65 70 75 80 Asn Asn Pro
Ala Trp Ala Val Gln Glu Glu Thr Arg Asp Arg Phe His 85 90 95 Leu
Leu Gly Asp Pro Gln Thr Lys Asn Cys Thr Leu Ser Ile Arg Asp 100 105
110 Ala Arg Met Ser Asp Ala Gly Arg Tyr Phe Phe Arg Met Glu Lys Gly
115 120 125 Asn Ile Lys Trp Asn Tyr Lys Tyr Asp Gln Leu Ser Val Asn
Val Thr 130 135 140 Tyr Pro Pro Gln Asn Leu Thr Val Thr Val Phe Gln
Gly Glu Gly Thr 145 150 155 160 Ala Ser Thr Ala Leu Gly Asn Ser Ser
Ser Leu Ser Val Leu Glu Gly 165 170 175 Gln Ser Leu Arg Leu Val Cys
Ala Val Asp Ser Asn Pro Pro Ala Arg 180 185 190 Leu Ser Trp Thr Trp
Arg Ser Leu Thr Leu Tyr Pro Ser Gln Pro Ser 195 200 205 Asn Pro Leu
Val Leu Glu Leu Gln Val His Leu Gly Asp Glu Gly Glu 210 215 220 Phe
Thr Cys Arg Ala Gln Asn Ser Leu Gly Ser Gln His Val Ser Leu 225 230
235 240 Asn Leu Ser Leu Gln Gln Glu Tyr Thr Gly Lys Met Lys Pro Val
Ser 245 250 255 Gly Val Leu Leu Gly Ala Val Gly Gly Thr Gly Ala Thr
Ala Leu Val 260 265 270 Phe Leu Ser Phe Cys Val Ile Phe Ile Val Val
Arg Ser Cys Arg Lys 275 280 285 Lys Ser Ala Arg Pro Ala Ala Asp Val
Gly Asp Ile Gly Met Lys Asp 290 295 300 Ala Asn Thr Ile Gln Gly Leu
Ser Leu Ser Gly 305 310 315 19 245 PRT Homo sapiens 19 Met Ala Glu
Ile His Thr Pro Tyr Ser Ser Leu Lys Lys Leu Leu Ser 1 5 10 15 Leu
Leu Asn Gly Phe Val Ala Val Ser Gly Ile Ile Leu Val Gly Leu 20 25
30 Gly Ile Gly Gly Lys Cys Gly Gly Ala Ser Leu Thr Asn Val Leu Gly
35 40 45 Leu Ser Ser Ala Tyr Leu Leu His Val Gly Asn Leu Cys Leu
Val Met 50 55 60 Gly Cys Ile Thr Val Leu Leu Gly Cys Ala Gly Trp
Tyr Gly Ala Thr 65 70 75 80 Lys Glu Ser Arg Gly Thr Leu Leu Phe Cys
Ile Leu Ser Met Val Ile 85 90 95 Val Leu Ile Met Glu Val Thr Ala
Ala Thr Val Val Leu Leu Phe Phe 100 105 110 Pro Ile Val Gly Asp Val
Ala Leu Glu His Thr Phe Val Thr Leu Arg 115 120 125 Lys Asn Tyr Arg
Gly Tyr Asn Glu Pro Asp Asp Tyr Ser Thr Gln Trp 130 135 140 Asn Leu
Val Met Glu Lys Leu Lys Cys Cys Gly Val Asn Asn Tyr Thr 145 150 155
160 Asp Phe Ser Gly Ser Ser Phe Glu Met Thr Thr Gly His Thr Tyr Pro
165 170 175 Arg Ser Cys Cys Lys Ser Ile Gly Ser Val Ser Cys Asp Gly
Arg Asp 180 185 190 Val Ser Pro Asn Val Ile His Gln Lys Gly Cys Phe
His Lys Leu Leu 195 200 205 Lys Ile Thr Lys Thr Gln Ser Phe Thr Leu
Ser Gly Ser Ser Leu Gly 210 215 220 Ala Ala Val Ile Gln Arg Trp Gly
Ser Arg Tyr Val Ala Gln Ala Gly 225 230 235 240 Leu Glu Leu Leu Ala
245 20 273 PRT Homo sapiens 20 Met Gln Cys Phe Ser Phe Ile Lys Thr
Met Met Ile Leu Phe Asn Leu 1 5 10 15 Leu Ile Phe Leu Cys Gly Ala
Ala Leu Leu Ala Val Gly Ile Trp Val 20 25 30 Ser Ile Asp Gly Ala
Ser Phe Leu Lys Ile Phe Gly Pro Leu Ser Ser 35 40 45 Ser Ala Met
Gln Phe Val Asn Val Gly Tyr Phe Leu Ile Ala Ala Gly 50 55 60 Val
Val Val Phe Ala Leu Gly Phe Leu Gly Cys Tyr Gly Ala Lys Thr 65 70
75 80 Glu Ser Lys Cys Ala Leu Val Thr Phe Phe Phe Ile Leu Leu Leu
Ile 85 90 95 Phe Ile Ala Glu Val Ala Ala Ala Val Val Ala Leu Val
Tyr Thr Thr 100 105 110 Met Ala Glu His Phe Leu Thr Leu Leu Val Val
Pro Ala Ile Lys Lys 115 120 125 Asp Tyr Gly Ser Gln Glu Asp Phe Thr
Gln Val Trp Asn Thr Thr Met 130 135 140 Lys Gly Leu Lys Cys Cys Gly
Phe Thr Asn Tyr Thr Asp Phe Glu Asp 145 150 155 160 Ser Pro Tyr Phe
Lys Glu Asn Ser Ala Phe Pro Pro Phe Cys Cys Asn 165 170 175 Asp Asn
Val Thr Asn Thr Ala Asn Glu Thr Cys Thr Lys Gln Lys Ala 180 185 190
His Asp Gln Lys Val Glu Gly Cys Phe Asn Gln Leu Leu Tyr Asp Ile 195
200 205 Arg Thr Asn Ala Val Thr Val Gly Gly Val Ala Ala Gly Ile Gly
Gly 210 215 220 Leu Glu Leu Ala Ala Met Asn Cys Val His Val Ser Val
Leu Gln Ser 225 230 235 240 Thr Ile Ser Pro Leu Leu Pro Leu Pro Leu
Leu Leu Pro His Gly Asn 245 250 255 Cys Glu Glu Ala Pro Trp Gln Ala
Ala Val Ile Gly Gly Gly Asp Arg 260 265 270 Ile 21 250 PRT Homo
sapiens 21 Met Asn Ser Met Thr Ser Ala Val Pro Val Ala Asn Ser Val
Leu Val 1 5 10 15 Val Ala Pro His Asn Gly Tyr Pro Val Thr Pro Gly
Ile Met Ser His 20 25 30 Val Pro Leu Tyr Pro Asn Ser Gln Pro Gln
Val His Leu Val Pro Gly 35 40 45 Asn Pro Pro Ser Leu Val Ser Asn
Val Asn Gly Gln Pro Val Gln Lys 50 55 60 Ala Leu Lys Glu Gly Lys
Thr Leu Gly Ala Ile Gln Ile Ile Ile Gly 65 70 75 80 Leu Ala His Ile
Gly Leu Gly Ser Ile Met Ala Thr Val Leu Val Gly 85 90 95 Glu Tyr
Leu Ser Ile Ser Phe Tyr Gly Gly Phe Pro Phe Trp Gly Gly 100 105 110
Leu Trp Phe Ile Ile Ser Gly Ser Leu Ser Val Ala Ala Glu Asn Gln 115
120 125 Pro Tyr Ser Tyr Cys Leu Leu Ser Gly Ser Leu Gly Leu Asn Ile
Val 130 135 140 Ser Ala Ile Cys Ser Ala Val Gly Val Ile Leu Phe Ile
Thr Asp Leu 145 150 155 160 Ser Ile Pro His Pro Tyr Ala Tyr Pro Asp
Tyr Tyr Pro Tyr Ala Trp 165 170 175 Gly Val Asn Pro Gly Met Ala Ile
Ser Gly Val Leu Leu Val Phe Cys 180 185 190 Leu Leu Glu Phe Gly Ile
Ala Cys Ala Ser Ser His Phe Gly Cys Gln 195 200 205 Leu Val Cys Cys
Gln Ser Ser Asn Val Ser Val Ile Tyr Pro Asn Ile 210 215 220 Tyr Ala
Ala Asn Pro Val Ile Thr Pro Glu Pro Val Thr Ser Pro Pro 225 230 235
240 Ser Tyr Ser Ser Glu Ile Gln Ala Asn Lys 245 250 22 245 PRT Homo
sapiens 22 Met Ala Ser Pro Ser Arg Arg Leu Gln Thr Lys Pro Val Ile
Thr Cys 1 5 10 15 Phe Lys Ser Val Leu Leu Ile Tyr Thr Phe Ile Phe
Trp Ile Thr Gly 20 25 30 Val Ile Leu Leu Ala Val Gly Ile Trp Gly
Lys Val Ser Leu Glu Asn 35 40 45 Tyr Phe Ser Leu Leu Asn Glu Lys
Ala Thr Asn Val Pro Phe Val Leu 50 55 60 Ile Ala Thr Gly Thr Val
Ile Ile Leu Leu Gly Thr Phe Gly Cys Phe 65 70 75 80 Ala Thr Cys Arg
Ala Ser Ala Trp Met Leu Lys Leu Tyr Ala Met Phe 85 90 95 Leu Thr
Leu Val Phe Leu Val Glu Leu Val Ala Ala Ile Val Gly Phe 100 105 110
Val Phe Arg His Glu Ile Lys Asn Ser Phe Lys Asn Asn Tyr Glu Lys 115
120 125 Ala Leu Lys Gln Tyr Asn Ser Thr Gly Asp Tyr Arg Ser His Ala
Val 130 135 140 Asp Lys Ile Gln Asn Thr Leu His Cys Cys Gly Val Thr
Asp Tyr Arg 145 150 155 160 Asp Trp Thr Asp Thr Asn Tyr Tyr Ser Glu
Lys Gly Phe Pro Lys Ser 165 170 175 Cys Cys Lys Leu Glu Asp Cys Thr
Pro Gln Arg Asp Ala Asp Lys Val 180 185 190 Asn Asn Glu Gly Cys Phe
Ile Lys Val Met Thr Ile Ile Glu Ser Glu 195 200 205 Met Gly Val Val
Ala Gly Ile Ser Phe Gly Val Ala Cys Phe Gln Leu 210 215 220 Ile Gly
Ile Phe Leu Ala Tyr Cys Leu Ser Arg Ala Ile Thr Asn Asn 225 230 235
240 Gln Tyr Glu Ile Val 245 23 253 PRT Homo sapiens 23 Met Gly Gln
Cys Gly Ile Thr Ser Ser Lys Thr Val Leu Val Phe Leu 1 5 10 15 Asn
Leu Ile Phe Trp Gly Ala Ala Gly Ile Leu Cys Tyr Val Gly Ala 20 25
30 Tyr Val Phe Ile Thr Tyr Asp Asp Tyr Asp His Phe Phe Glu Asp Val
35 40 45 Tyr Thr Leu Ile Pro Ala Val Val Ile Ile Ala Val Gly Ala
Leu Leu 50 55 60 Phe Ile Ile Gly Leu Ile Gly Cys Cys Ala Thr Ile
Arg Glu Ser Arg 65 70 75 80 Cys Gly Leu Ala Thr Phe Val Ile Ile Leu
Leu Leu Val Phe Val Thr 85 90 95 Glu Val Val Val Val Val Leu Gly
Tyr Val Tyr Arg Ala Lys Val Glu 100 105 110 Asn Glu Val Asp Arg Ser
Ile Gln Lys Val Tyr Lys Thr Tyr Asn Gly 115 120 125 Thr Asn Pro Asp
Ala Ala Ser Arg Ala Ile Asp Tyr Val Gln Arg Gln 130 135 140 Leu His
Cys Cys Gly Ile His Asn Tyr Ser Asp Trp Glu Asn Thr Asp 145 150 155
160 Trp Phe Lys Glu Thr Lys Asn Gln Ser Val Pro Leu Ser Cys Cys Arg
165 170 175 Glu Thr Ala Ser Asn Cys Asn Gly Ser Leu Ala His Pro Ser
Asp Leu 180 185 190 Tyr Ala Glu Gly Cys Glu Ala Leu Val Val Lys Lys
Leu Gln Glu Ile 195 200 205 Met Met His Val Ile Trp Ala Ala Leu Ala
Phe Ala Ala Ile Gln Leu 210 215 220 Leu Gly Met Leu Cys Ala Cys Ile
Val Leu Cys Arg Arg Ser Arg Asp 225 230 235 240 Pro Ala Tyr Glu Leu
Phe Ile Thr Gly Gly Thr Tyr Ala 245 250 24 238 PRT Homo sapiens 24
Met Ala Arg Ala Cys Leu Gln Ala Val Lys Tyr Leu Met Phe Ala Phe 1 5
10 15 Asn Leu Phe Phe Trp Leu Gly Gly Cys Gly Val Leu Gly Val Gly
Ile 20 25 30 Trp Leu Ala Ala Thr Gln Gly Ser Phe Ala Thr Leu Ser
Ser Ser Phe 35 40 45 Pro Ser Leu Ser Ala Ala Asn Leu Leu Ile Ile
Thr Gly Ala Phe Val 50 55 60 Met Ala Ile Gly Phe Val Gly Cys Leu
Gly Ala Ile Lys Glu Asn Lys 65 70 75 80 Cys Leu Leu Leu Thr Phe Phe
Leu Leu Leu Leu Leu Val Phe Leu Leu 85 90 95 Glu Ala Thr Ile Ala
Ile Leu Phe Phe Ala Tyr Thr Asp Lys Ile Asp 100 105 110 Arg Tyr Ala
Gln Gln Asp Leu Lys Lys Gly Leu His Leu Tyr Gly Thr 115 120 125 Gln
Gly Asn Val Gly Leu Thr Asn Ala Trp Ser Ile Ile Gln Thr Asp 130 135
140 Phe Arg Cys Cys Gly Val Ser Asn Tyr Thr Asp Trp Phe Glu Val Tyr
145 150 155 160 Asn Ala Thr Arg Val Pro Asp Ser Cys Cys Leu Glu Phe
Ser Glu Ser 165 170 175 Cys Gly Leu His Ala Pro Gly Thr Trp Trp Lys
Ala Pro Cys Tyr Glu 180 185 190 Thr Val Lys Val Trp Leu Gln Glu Asn
Leu Leu Ala Val Gly Ile Phe 195 200 205 Gly Leu Cys Thr Ala Leu Val
Gln Ile Leu Gly Leu Thr Phe Ala Met 210 215 220 Thr Met Tyr Cys Gln
Val Val Lys Ala Asp Thr Tyr Cys Ala 225 230 235 25 221 PRT Homo
sapiens 25 Met Gly Arg Phe Arg
Gly Gly Leu Arg Cys Ile Lys Tyr Leu Leu Leu 1 5 10 15 Gly Phe Asn
Leu Leu Phe Trp Leu Ala Gly Ser Ala Val Ile Ala Phe 20 25 30 Gly
Leu Trp Phe Arg Phe Gly Gly Ala Ile Lys Glu Leu Ser Ser Glu 35 40
45 Asp Lys Ser Pro Glu Tyr Phe Tyr Val Gly Leu Tyr Val Leu Val Gly
50 55 60 Ala Gly Ala Leu Met Met Ala Val Gly Phe Phe Gly Cys Cys
Gly Ala 65 70 75 80 Met Arg Glu Ser Gln Cys Val Leu Gly Ser Phe Phe
Thr Cys Leu Leu 85 90 95 Val Ile Phe Ala Ala Glu Val Thr Thr Gly
Val Phe Ala Phe Ile Gly 100 105 110 Lys Gly Val Ala Ile Arg His Val
Gln Thr Met Tyr Glu Glu Ala Tyr 115 120 125 Asn Asp Tyr Leu Lys Asp
Arg Gly Lys Gly Asn Gly Thr Leu Ile Thr 130 135 140 Phe His Ser Thr
Phe Gln Cys Cys Gly Lys Glu Ser Ser Glu Gln Val 145 150 155 160 Gln
Pro Thr Cys Pro Lys Glu Leu Leu Gly His Lys Asn Cys Ile Asp 165 170
175 Glu Ile Glu Thr Ile Ile Ser Val Lys Leu Gln Leu Ile Gly Ile Val
180 185 190 Gly Ile Gly Ile Ala Gly Leu Thr Ile Phe Gly Met Ile Phe
Ser Met 195 200 205 Val Leu Cys Cys Ala Ile Arg Asn Ser Arg Asp Val
Ile 210 215 220 26 252 PRT Homo sapiens 26 Met Ala His Tyr Lys Thr
Glu Gln Asp Asp Trp Leu Ile Ile Tyr Leu 1 5 10 15 Lys Tyr Leu Leu
Phe Val Phe Asn Phe Phe Phe Trp Val Gly Gly Ala 20 25 30 Ala Val
Leu Ala Val Gly Ile Trp Thr Leu Val Glu Lys Ser Gly Tyr 35 40 45
Leu Ser Val Leu Ala Ser Ser Thr Phe Ala Ala Ser Ala Tyr Ile Leu 50
55 60 Ile Phe Ala Gly Val Leu Val Met Val Thr Gly Phe Leu Gly Phe
Gly 65 70 75 80 Ala Ile Leu Trp Glu Arg Lys Gly Cys Leu Ser Thr Tyr
Phe Cys Leu 85 90 95 Leu Leu Val Ile Phe Leu Val Glu Leu Val Ala
Gly Val Leu Ala His 100 105 110 Val Tyr Tyr Gln Arg Leu Ser Asp Glu
Leu Lys Gln His Leu Asn Arg 115 120 125 Thr Leu Ala Glu Asn Tyr Gly
Gln Pro Glu His Ala Asp His Ala Ser 130 135 140 Val Asp Arg Leu Gln
Gln Asp Phe Lys Cys Cys Gly Ser Asn Ser Ser 145 150 155 160 Ala Asp
Trp Gln His Ser Thr Tyr Ile Leu Leu Arg Glu Ala Glu Gly 165 170 175
Arg Gln Val Pro Asp Ser Cys Cys Lys Thr Val Val Ala Arg Cys Gly 180
185 190 Gln Arg Ala His Pro Ser Asn Ile Tyr Lys Val Glu Gly Gly Cys
Leu 195 200 205 Thr Lys Leu Glu Gln Phe Leu Ala Asp His Leu Leu Leu
Met Gly Ala 210 215 220 Val Gly Ile Gly Val Ala Cys Leu Gln Ile Cys
Gly Met Val Leu Thr 225 230 235 240 Cys Cys Leu His Gln Arg Leu Gln
Arg His Phe Tyr 245 250 27 197 PRT Homo sapiens 27 Met Cys Thr Gly
Lys Cys Ala Arg Cys Val Gly Leu Ser Leu Ile Thr 1 5 10 15 Leu Cys
Leu Val Cys Ile Val Ala Asn Ala Leu Leu Leu Val Pro Asn 20 25 30
Gly Glu Thr Ser Trp Thr Asn Thr Asn His Leu Ser Leu Gln Val Trp 35
40 45 Leu Met Gly Gly Phe Ile Gly Gly Gly Leu Met Val Leu Cys Pro
Gly 50 55 60 Ile Ala Ala Val Arg Ala Gly Gly Lys Gly Cys Cys Gly
Ala Gly Cys 65 70 75 80 Cys Gly Asn Arg Cys Arg Met Leu Arg Ser Val
Phe Ser Ser Ala Phe 85 90 95 Gly Val Leu Gly Ala Ile Tyr Cys Leu
Ser Val Ser Gly Ala Gly Leu 100 105 110 Arg Asn Gly Pro Arg Cys Leu
Met Asn Gly Glu Trp Gly Tyr His Phe 115 120 125 Glu Asp Thr Ala Gly
Ala Tyr Leu Leu Asn Arg Thr Leu Trp Asp Arg 130 135 140 Cys Glu Ala
Pro Pro Arg Val Val Pro Trp Asn Val Thr Leu Phe Ser 145 150 155 160
Leu Leu Val Ala Ala Ser Cys Leu Glu Ile Val Leu Cys Gly Ile Gln 165
170 175 Leu Val Asn Ala Thr Ile Gly Val Phe Cys Gly Asp Cys Arg Lys
Lys 180 185 190 Gln Asp Thr Pro His 195 28 201 PRT Homo sapiens 28
Met Gly Ser Arg Lys Cys Gly Gly Cys Leu Ser Cys Leu Leu Ile Pro 1 5
10 15 Leu Ala Leu Trp Ser Ile Ile Val Asn Ile Leu Leu Tyr Phe Pro
Asn 20 25 30 Gly Gln Thr Ser Tyr Ala Ser Ser Asn Lys Leu Thr Asn
Tyr Val Trp 35 40 45 Tyr Phe Glu Gly Ile Cys Phe Ser Gly Ile Met
Met Leu Ile Val Thr 50 55 60 Thr Val Leu Leu Val Leu Glu Asn Asn
Asn Asn Tyr Lys Cys Cys Gln 65 70 75 80 Ser Glu Asn Cys Ser Lys Lys
Tyr Val Thr Leu Leu Ser Ile Ile Phe 85 90 95 Ser Ser Leu Gly Ile
Ala Phe Ser Gly Tyr Cys Leu Val Ile Ser Ala 100 105 110 Leu Gly Leu
Val Gln Gly Pro Tyr Cys Arg Thr Leu Asp Gly Trp Glu 115 120 125 Tyr
Ala Phe Glu Gly Thr Ala Gly Arg Phe Leu Thr Asp Ser Ser Ile 130 135
140 Trp Ile Gln Cys Leu Glu Pro Ala His Val Val Glu Trp Asn Ile Ile
145 150 155 160 Leu Phe Ser Ile Leu Ile Thr Leu Ser Gly Leu Gln Val
Ile Ile Cys 165 170 175 Leu Ile Arg Val Val Met Gln Leu Ser Lys Ile
Leu Cys Gly Ser Tyr 180 185 190 Ser Val Ile Phe Gln Pro Gly Ile Ile
195 200 29 133 PRT Homo sapiens 29 Met Ser Pro Arg Leu Glu Val Pro
Cys Ser His Ala Leu Pro Gln Gly 1 5 10 15 Leu Ser Pro Gly Gln Val
Ile Ile Val Arg Gly Leu Val Leu Gln Glu 20 25 30 Pro Lys His Phe
Thr Val Ser Leu Arg Asp Gln Ala Ala His Ala Pro 35 40 45 Val Thr
Leu Arg Ala Ser Phe Ala Asp Arg Thr Leu Ala Trp Ile Ser 50 55 60
Arg Trp Gly Gln Lys Lys Leu Ile Ser Ala Pro Phe Leu Phe Tyr Pro 65
70 75 80 Gln Arg Phe Phe Glu Val Leu Leu Leu Phe Gln Glu Gly Gly
Leu Lys 85 90 95 Leu Ala Leu Asn Gly Gln Gly Leu Gly Ala Thr Ser
Met Asn Gln Gln 100 105 110 Ala Leu Glu Gln Leu Arg Glu Leu Arg Ile
Ser Gly Ser Val Gln Leu 115 120 125 Tyr Cys Val His Ser 130 30 70
PRT Homo sapiens 30 Met Arg Thr Ala Leu Leu Leu Leu Ala Ala Leu Ala
Val Ala Thr Gly 1 5 10 15 Pro Ala Leu Thr Leu Arg Cys His Val Cys
Thr Ser Ser Ser Asn Cys 20 25 30 Lys His Ser Val Val Cys Pro Ala
Ser Ser Arg Phe Cys Lys Thr Thr 35 40 45 Asn Thr Val Glu Pro Leu
Arg Ala Ser Pro Lys Val Trp Asp Gln Val 50 55 60 Gln Val Gly Met
Glu Cys 65 70 31 299 PRT Homo sapiens 31 Met Ala Gln Asn Leu Lys
Asp Leu Ala Gly Arg Leu Pro Ala Gly Pro 1 5 10 15 Arg Gly Met Gly
Thr Ala Leu Lys Leu Leu Leu Gly Ala Gly Ala Val 20 25 30 Ala Tyr
Gly Val Arg Glu Ser Val Phe Thr Val Glu Gly Gly His Arg 35 40 45
Ala Ile Phe Phe Asn Arg Ile Gly Gly Val Gln Gln Asp Thr Ile Leu 50
55 60 Ala Glu Gly Leu His Phe Arg Ile Pro Trp Phe Gln Tyr Pro Ile
Ile 65 70 75 80 Tyr Asp Ile Arg Ala Arg Pro Arg Lys Ile Ser Ser Pro
Thr Gly Ser 85 90 95 Lys Asp Leu Gln Met Val Asn Ile Ser Leu Arg
Val Leu Ser Arg Pro 100 105 110 Asn Ala Gln Glu Leu Pro Ser Met Tyr
Gln Arg Leu Gly Leu Asp Tyr 115 120 125 Glu Glu Arg Val Leu Pro Ser
Ile Val Asn Glu Val Leu Lys Ser Val 130 135 140 Val Ala Lys Phe Asn
Ala Ser Gln Leu Ile Thr Gln Arg Ala Gln Val 145 150 155 160 Ser Leu
Leu Ile Arg Arg Glu Leu Thr Glu Arg Ala Lys Asp Phe Ser 165 170 175
Leu Ile Leu Asp Asp Val Ala Ile Thr Glu Leu Ser Phe Ser Arg Glu 180
185 190 Tyr Thr Ala Ala Val Glu Ala Lys Gln Val Ala Gln Gln Glu Ala
Gln 195 200 205 Arg Ala Gln Phe Leu Val Glu Lys Ala Lys Gln Glu Gln
Arg Gln Lys 210 215 220 Ile Val Gln Ala Glu Gly Glu Ala Glu Ala Ala
Lys Met Leu Gly Glu 225 230 235 240 Ala Leu Ser Lys Asn Pro Gly Tyr
Ile Lys Leu Arg Lys Ile Arg Ala 245 250 255 Ala Gln Asn Ile Ser Lys
Thr Ile Ala Thr Ser Gln Asn Arg Ile Tyr 260 265 270 Leu Thr Ala Asp
Asn Leu Val Leu Asn Leu Gln Asp Glu Ser Phe Thr 275 280 285 Arg Gly
Ser Asp Ser Leu Ile Lys Gly Lys Lys 290 295 32 168 PRT Homo sapiens
32 Met Asp Cys Glu Ile Lys Gly Arg Pro Cys Cys Ile Gly Thr Lys Gly
1 5 10 15 Ser Cys Glu Ile Thr Thr Arg Glu Tyr Cys Glu Phe Met His
Gly Tyr 20 25 30 Phe His Glu Glu Ala Thr Leu Cys Ser Gln Val Arg
Arg Gly Arg Pro 35 40 45 Gly Val Val Glu Glu Arg Thr Leu Gly Met
Ala Ala Cys Trp Gly Arg 50 55 60 Gly Ser Arg Thr Pro Ser His Val
Gly Ala Ser Asp Ser Gly Cys Phe 65 70 75 80 Trp Gly Ala Glu His His
Met Pro Ile Pro Arg Cys Thr Val Leu Asp 85 90 95 Lys Val Cys Trp
Ala Ala Ala Phe Leu Asn Pro Glu Val Pro Asp Gln 100 105 110 Phe Tyr
Arg Ser Gly Cys Leu Phe Ser Tyr Met Leu Gly Lys Arg Ser 115 120 125
Ser Met Pro Pro Asn Pro Thr Pro Val Met Asp Thr Gln Ala Asp Pro 130
135 140 Trp Gly Lys Val Pro Gly Pro Gly Tyr Gly Arg Ser Asn Leu Pro
Lys 145 150 155 160 Thr Thr Ala Pro Glu Val Ser Gly 165 33 551 PRT
Homo sapiens 33 Met Leu Pro Leu Leu Leu Leu Pro Leu Leu Trp Gly Gly
Ser Leu Gln 1 5 10 15 Glu Lys Pro Val Tyr Glu Leu Gln Val Gln Lys
Ser Val Thr Val Gln 20 25 30 Glu Gly Leu Cys Val Leu Val Pro Cys
Ser Phe Ser Tyr Pro Trp Arg 35 40 45 Ser Trp Tyr Ser Ser Pro Pro
Leu Tyr Val Tyr Trp Phe Arg Asp Gly 50 55 60 Glu Ile Pro Tyr Tyr
Ala Glu Val Val Ala Thr Asn Asn Pro Asp Arg 65 70 75 80 Arg Val Lys
Pro Glu Thr Gln Gly Arg Phe Arg Leu Leu Gly Asp Val 85 90 95 Gln
Lys Lys Asn Cys Ser Leu Ser Ile Gly Asp Ala Arg Met Glu Asp 100 105
110 Thr Gly Ser Tyr Phe Phe Arg Val Glu Arg Gly Arg Asp Val Lys Tyr
115 120 125 Ser Tyr Gln Gln Asn Lys Leu Asn Leu Glu Val Thr Ala Leu
Ile Glu 130 135 140 Lys Pro Asp Ile His Phe Leu Glu Pro Leu Glu Ser
Gly Arg Pro Thr 145 150 155 160 Arg Leu Ser Cys Ser Leu Pro Gly Ser
Cys Glu Ala Gly Pro Pro Leu 165 170 175 Thr Phe Ser Trp Thr Gly Asn
Ala Leu Ser Pro Leu Asp Pro Glu Thr 180 185 190 Thr Arg Ser Ser Glu
Leu Thr Leu Thr Pro Arg Pro Glu Asp His Gly 195 200 205 Thr Asn Leu
Thr Cys Gln Met Lys Arg Gln Gly Ala Gln Val Thr Thr 210 215 220 Glu
Arg Thr Val Gln Leu Asn Val Ser Tyr Ala Pro Gln Thr Ile Thr 225 230
235 240 Ile Phe Arg Asn Gly Ile Ala Leu Glu Ile Leu Gln Asn Thr Ser
Tyr 245 250 255 Leu Pro Val Leu Glu Gly Gln Ala Leu Arg Leu Leu Cys
Asp Ala Pro 260 265 270 Ser Asn Pro Pro Ala His Leu Ser Trp Phe Gln
Gly Ser Pro Ala Leu 275 280 285 Asn Ala Thr Pro Ile Ser Asn Thr Gly
Ile Leu Glu Leu Arg Arg Val 290 295 300 Arg Ser Ala Glu Glu Gly Gly
Phe Thr Cys Arg Ala Gln His Pro Leu 305 310 315 320 Gly Phe Leu Gln
Ile Phe Leu Asn Leu Ser Val Tyr Ser Leu Pro Gln 325 330 335 Leu Leu
Gly Pro Ser Cys Ser Trp Glu Ala Glu Gly Leu His Cys Arg 340 345 350
Cys Ser Phe Arg Ala Arg Pro Ala Pro Ser Leu Cys Trp Arg Leu Glu 355
360 365 Glu Lys Pro Leu Glu Gly Asn Ser Ser Gln Gly Ser Phe Lys Val
Asn 370 375 380 Ser Ser Ser Ala Gly Pro Trp Ala Asn Ser Ser Leu Ile
Leu His Gly 385 390 395 400 Gly Leu Ser Ser Asp Leu Lys Val Ser Cys
Lys Ala Trp Asn Ile Tyr 405 410 415 Gly Ser Gln Ser Gly Ser Val Leu
Leu Leu Gln Gly Arg Ser Asn Leu 420 425 430 Gly Thr Gly Val Val Pro
Ala Ala Leu Gly Gly Ala Gly Val Met Ala 435 440 445 Leu Leu Cys Ile
Cys Leu Cys Leu Ile Phe Phe Leu Ile Val Lys Ala 450 455 460 Arg Arg
Lys Gln Ala Ala Gly Arg Pro Glu Lys Met Asp Asp Glu Asp 465 470 475
480 Pro Ile Met Gly Thr Ile Thr Ser Gly Ser Arg Lys Lys Pro Trp Pro
485 490 495 Asp Ser Pro Gly Asp Gln Ala Ser Pro Pro Gly Asp Ala Pro
Pro Leu 500 505 510 Glu Glu Gln Lys Glu Leu His Tyr Ala Ser Leu Ser
Phe Ser Glu Met 515 520 525 Lys Ser Arg Glu Pro Lys Asp Gln Glu Ala
Pro Ser Thr Thr Glu Tyr 530 535 540 Ser Glu Ile Lys Thr Ser Lys 545
550 34 219 PRT Rattus sp. 34 Met Gly Met Ser Ser Leu Lys Leu Leu
Lys Tyr Val Leu Phe Phe Phe 1 5 10 15 Asn Phe Leu Phe Trp Val Cys
Gly Cys Cys Ile Leu Gly Phe Gly Ile 20 25 30 His Leu Leu Val Gln
Asn Thr Tyr Gly Ile Leu Phe Arg Asn Leu Pro 35 40 45 Phe Leu Thr
Leu Gly Asn Val Leu Val Ile Val Gly Ser Ile Ile Met 50 55 60 Val
Val Ala Phe Leu Gly Cys Met Gly Ser Ile Lys Glu Asn Lys Cys 65 70
75 80 Leu Leu Met Ser Phe Phe Val Leu Leu Leu Leu Ile Leu Leu Ala
Glu 85 90 95 Val Thr Leu Ala Ile Leu Leu Phe Val Tyr Glu Lys Lys
Ile Asn Thr 100 105 110 Leu Val Ala Glu Gly Leu Asn Asp Ser Ile Gln
His Tyr His Ser Asp 115 120 125 Asn Ser Thr Arg Met Ala Trp Asp Phe
Ile Gln Ser Gln Leu Gln Cys 130 135 140 Cys Gly Val Asn Gly Ser Ser
Asp Trp Ile Ser Gly Pro Pro Ser Ser 145 150 155 160 Cys Pro Ser Gly
Ala Asp Val Gln Gly Cys Tyr Lys Lys Gly Gln Ala 165 170 175 Trp Phe
His Ser Asn Phe Leu Tyr Ile Gly Ile Val Thr Ile Cys Val 180 185 190
Cys Val Ile Gln Val Leu Gly Met Ser Phe Ala Leu Thr Leu Asn Cys 195
200 205 Gln Ile Asp Lys Thr Ser Gln Ala Leu Gly Leu 210 215 35 253
PRT Homo sapiens 35 Met Gly Glu Phe Asn Glu Lys Lys Thr Thr Cys Gly
Thr Val Cys Leu 1 5 10 15 Lys Tyr Leu Leu Phe Thr Tyr Asn Cys Cys
Phe Trp Leu Ala Gly Leu 20 25 30 Ala Val Met Ala Val Gly Ile Trp
Thr Leu Ala Leu Lys Ser Asp Tyr 35 40 45 Ile Ser Leu Leu Ala Ser
Gly Thr Tyr Leu Ala Thr Ala Tyr Ile Leu 50 55 60 Val Val Ala Gly
Thr Val Val Met Val Thr Gly Val Leu Gly Cys Cys 65 70 75 80 Ala Thr
Phe Lys Glu Arg Arg Asn Leu Leu Arg Leu Tyr Phe Ile Leu 85 90 95
Leu Leu Ile Ile Phe Leu Leu Glu Ile Ile Ala Gly Ile Leu Ala Tyr 100
105 110 Ala Tyr Tyr Gln
Gln Leu Asn Thr Glu Leu Lys Glu Asn Leu Lys Asp 115 120 125 Thr Met
Thr Lys Arg Tyr His Gln Pro Gly His Glu Ala Val Thr Ser 130 135 140
Ala Val Asp Gln Leu Gln Gln Glu Phe His Cys Cys Gly Ser Asn Asn 145
150 155 160 Ser Gln Asp Trp Arg Asp Ser Glu Trp Ile Arg Ser Gln Glu
Ala Gly 165 170 175 Gly Arg Val Val Pro Asp Ser Cys Cys Lys Thr Val
Val Ala Leu Cys 180 185 190 Gly Gln Arg Asp His Ala Ser Asn Ile Tyr
Lys Val Glu Gly Gly Cys 195 200 205 Ile Thr Lys Leu Glu Thr Phe Ile
Gln Glu His Leu Arg Val Ile Gly 210 215 220 Ala Val Gly Ile Gly Ile
Ala Cys Val Gln Val Phe Gly Met Ile Phe 225 230 235 240 Thr Cys Cys
Leu Tyr Arg Ser Leu Lys Leu Glu His Tyr 245 250 36 238 PRT Homo
sapiens 36 Met Ala Arg Ala Cys Leu Gln Ala Val Lys Tyr Leu Met Phe
Ala Phe 1 5 10 15 Asn Leu Leu Phe Trp Leu Gly Gly Cys Gly Val Leu
Gly Val Gly Ile 20 25 30 Trp Leu Ala Ala Thr Gln Gly Ser Phe Ala
Thr Leu Ser Ser Ser Phe 35 40 45 Pro Ser Leu Ser Ala Ala Asn Leu
Leu Ile Ile Thr Gly Ala Phe Val 50 55 60 Met Ala Ile Gly Phe Val
Gly Cys Leu Gly Ala Ile Lys Glu Asn Lys 65 70 75 80 Cys Leu Leu Leu
Thr Phe Phe Leu Leu Leu Leu Leu Val Phe Leu Leu 85 90 95 Glu Ala
Thr Ile Ala Ile Leu Phe Phe Ala Tyr Thr Asp Lys Ile Asp 100 105 110
Arg Tyr Ala Gln Gln Asp Leu Lys Lys Gly Leu His Leu Tyr Gly Thr 115
120 125 Gln Gly Asn Val Gly Leu Thr Asn Ala Trp Ser Ile Ile Gln Thr
Asp 130 135 140 Phe Arg Cys Cys Gly Val Ser Asn Tyr Thr Asp Trp Phe
Glu Val Tyr 145 150 155 160 Asn Ala Thr Arg Val Pro Asp Ser Cys Cys
Leu Glu Phe Ser Glu Ser 165 170 175 Cys Gly Leu His Ala Pro Gly Thr
Trp Trp Lys Ala Pro Cys Tyr Glu 180 185 190 Thr Val Lys Val Trp Leu
Gln Glu Asn Leu Leu Ala Val Gly Ile Phe 195 200 205 Gly Leu Cys Thr
Ala Leu Val Gln Ile Leu Gly Leu Thr Phe Ala Met 210 215 220 Thr Met
Tyr Cys Gln Val Val Lys Ala Asp Thr Tyr Cys Ala 225 230 235 37 244
PRT Homo sapiens 37 Met Glu Thr Lys Pro Val Ile Thr Cys Leu Lys Thr
Leu Leu Ile Ile 1 5 10 15 Tyr Ser Phe Val Phe Trp Ile Thr Gly Val
Ile Leu Leu Ala Val Gly 20 25 30 Val Trp Gly Lys Leu Thr Leu Gly
Thr Tyr Ile Ser Leu Ile Ala Glu 35 40 45 Asn Ser Thr Asn Ala Pro
Tyr Val Leu Ile Gly Thr Gly Thr Thr Ile 50 55 60 Val Val Phe Gly
Leu Phe Gly Cys Phe Ala Thr Cys Arg Gly Ser Pro 65 70 75 80 Trp Met
Leu Lys Leu Tyr Ala Met Phe Leu Ser Leu Val Phe Leu Ala 85 90 95
Glu Leu Val Ala Gly Ile Ser Gly Phe Val Phe Arg His Glu Ile Lys 100
105 110 Asp Thr Phe Leu Arg Thr Tyr Thr Asp Ala Met Gln Thr Tyr Asn
Gly 115 120 125 Asn Asp Glu Arg Ser Arg Ala Val Asp His Val Gln Arg
Ser Leu Ser 130 135 140 Cys Cys Gly Val Gln Asn Tyr Thr Asn Trp Ser
Thr Ser Pro Tyr Phe 145 150 155 160 Leu Glu His Gly Ile Pro Pro Ser
Cys Cys Met Asn Glu Thr Asp Cys 165 170 175 Asn Pro Gln Asp Leu His
Asn Leu Thr Val Ala Ala Thr Lys Val Asn 180 185 190 Gln Lys Gly Cys
Tyr Asp Leu Val Thr Ser Phe Met Glu Thr Asn Met 195 200 205 Gly Ile
Ile Ala Gly Val Ala Phe Gly Ile Ala Phe Ser Gln Leu Ile 210 215 220
Gly Met Leu Leu Ala Cys Cys Leu Ser Arg Phe Ile Thr Ala Asn Gln 225
230 235 240 Tyr Glu Met Val 38 297 PRT Homo sapiens 38 Met Thr Thr
Pro Arg Asn Ser Val Asn Gly Thr Phe Pro Ala Glu Pro 1 5 10 15 Met
Lys Gly Pro Ile Ala Met Gln Ser Gly Pro Lys Pro Leu Phe Arg 20 25
30 Arg Met Ser Ser Leu Val Gly Pro Thr Gln Ser Phe Phe Met Arg Glu
35 40 45 Ser Lys Thr Leu Gly Ala Val Gln Ile Met Asn Gly Leu Phe
His Ile 50 55 60 Ala Leu Gly Gly Leu Leu Met Ile Pro Ala Gly Ile
Tyr Ala Pro Ile 65 70 75 80 Cys Val Thr Val Trp Tyr Pro Leu Trp Gly
Gly Ile Met Tyr Ile Ile 85 90 95 Ser Gly Ser Leu Leu Ala Ala Thr
Glu Lys Asn Ser Arg Lys Cys Leu 100 105 110 Val Lys Gly Lys Met Ile
Met Asn Ser Leu Ser Leu Phe Ala Ala Ile 115 120 125 Ser Gly Met Ile
Leu Ser Ile Met Asp Ile Leu Asn Ile Lys Ile Ser 130 135 140 His Phe
Leu Lys Met Glu Ser Leu Asn Phe Ile Arg Ala His Thr Pro 145 150 155
160 Tyr Ile Asn Ile Tyr Asn Cys Glu Pro Ala Asn Pro Ser Glu Lys Asn
165 170 175 Ser Pro Ser Thr Gln Tyr Cys Tyr Ser Ile Gln Ser Leu Phe
Leu Gly 180 185 190 Ile Leu Ser Val Met Leu Ile Phe Ala Phe Phe Gln
Glu Leu Val Ile 195 200 205 Ala Gly Ile Val Glu Asn Glu Trp Lys Arg
Thr Cys Ser Arg Pro Lys 210 215 220 Ser Asn Ile Val Leu Leu Ser Ala
Glu Glu Lys Lys Glu Gln Thr Ile 225 230 235 240 Glu Ile Lys Glu Glu
Val Val Gly Leu Thr Glu Thr Ser Ser Gln Pro 245 250 255 Lys Asn Glu
Glu Asp Ile Glu Ile Ile Pro Ile Gln Glu Glu Glu Glu 260 265 270 Glu
Glu Thr Glu Thr Asn Phe Pro Glu Pro Pro Gln Asp Gln Glu Ser 275 280
285 Ser Pro Ile Glu Asn Asp Ser Ser Pro 290 295 39 228 PRT Homo
sapiens 39 Met Pro Val Lys Gly Gly Thr Lys Cys Ile Lys Tyr Leu Leu
Phe Gly 1 5 10 15 Phe Asn Phe Ile Phe Trp Leu Ala Gly Ile Ala Val
Leu Ala Ile Gly 20 25 30 Leu Trp Leu Arg Phe Asp Ser Gln Thr Lys
Ser Ile Phe Glu Gln Glu 35 40 45 Thr Asn Asn Asn Asn Ser Ser Phe
Tyr Thr Gly Val Tyr Ile Leu Ile 50 55 60 Gly Ala Gly Ala Leu Met
Met Leu Val Gly Phe Leu Gly Cys Cys Gly 65 70 75 80 Ala Val Gln Glu
Ser Gln Cys Met Leu Gly Leu Phe Phe Gly Phe Leu 85 90 95 Leu Val
Ile Phe Ala Ile Glu Ile Ala Ala Ala Ile Trp Gly Tyr Ser 100 105 110
His Lys Asp Glu Val Ile Lys Glu Val Gln Glu Phe Tyr Lys Asp Thr 115
120 125 Tyr Asn Lys Leu Lys Thr Lys Asp Glu Pro Gln Arg Glu Thr Leu
Lys 130 135 140 Ala Ile His Tyr Ala Leu Asn Cys Cys Gly Leu Ala Gly
Gly Val Glu 145 150 155 160 Gln Phe Ile Ser Asp Ile Cys Pro Lys Lys
Asp Val Leu Glu Thr Phe 165 170 175 Thr Val Lys Ser Cys Pro Asp Ala
Ile Lys Glu Val Phe Asp Asn Lys 180 185 190 Phe His Ile Ile Gly Ala
Val Gly Ile Gly Ile Ala Val Val Met Ile 195 200 205 Phe Gly Met Ile
Phe Ser Met Ile Leu Cys Cys Ala Ile Arg Arg Asn 210 215 220 Arg Glu
Met Val 225 40 197 PRT Homo sapiens 40 Met Cys Thr Gly Lys Cys Ala
Arg Cys Val Gly Leu Ser Leu Ile Thr 1 5 10 15 Leu Cys Phe Val Cys
Ile Val Ala Asn Ala Leu Leu Leu Val Pro Asn 20 25 30 Gly Glu Thr
Ser Trp Thr Asn Thr Asn His Leu Ser Leu Gln Val Trp 35 40 45 Leu
Met Gly Gly Phe Ile Gly Gly Gly Leu Met Val Leu Cys Pro Gly 50 55
60 Ile Ala Ala Val Arg Ala Gly Gly Lys Gly Cys Cys Gly Ala Gly Cys
65 70 75 80 Cys Gly Asn Arg Cys Arg Met Leu Arg Ser Val Phe Ser Ser
Ala Phe 85 90 95 Gly Val Leu Gly Ala Ile Tyr Cys Leu Ser Val Ser
Gly Ala Gly Leu 100 105 110 Arg Asn Gly Pro Arg Cys Leu Met Asn Gly
Glu Trp Gly Tyr His Phe 115 120 125 Glu Asp Thr Ala Gly Ala Tyr Leu
Leu Asn Arg Thr Leu Trp Asp Arg 130 135 140 Cys Glu Ala Pro Pro Arg
Val Val Pro Trp Asn Val Thr Leu Phe Ser 145 150 155 160 Leu Leu Val
Ala Ala Ser Cys Leu Glu Ile Val Leu Cys Gly Ile Gln 165 170 175 Leu
Val Asn Ala Thr Ile Gly Val Phe Cys Gly Asp Cys Arg Lys Lys 180 185
190 Gln Asp Thr Pro His 195 41 202 PRT Homo sapiens 41 Met Cys Tyr
Gly Lys Cys Ala Arg Cys Ile Gly His Ser Leu Val Gly 1 5 10 15 Leu
Ala Leu Leu Cys Ile Ala Ala Asn Ile Leu Leu Tyr Phe Pro Asn 20 25
30 Gly Glu Thr Lys Tyr Ala Ser Glu Asn His Leu Ser Arg Phe Val Trp
35 40 45 Phe Phe Ser Gly Ile Val Gly Gly Gly Leu Leu Met Leu Leu
Pro Ala 50 55 60 Phe Val Phe Ile Gly Leu Glu Gln Asp Asp Cys Cys
Gly Cys Cys Gly 65 70 75 80 His Glu Asn Cys Gly Lys Arg Cys Ala Met
Leu Ser Ser Val Leu Ala 85 90 95 Ala Leu Ile Gly Ile Ala Gly Ser
Gly Tyr Cys Val Ile Val Ala Ala 100 105 110 Leu Gly Leu Ala Glu Gly
Pro Leu Cys Leu Asp Ser Leu Gly Gln Trp 115 120 125 Asn Tyr Thr Phe
Ala Ser Thr Glu Gly Gln Tyr Leu Leu Asp Thr Ser 130 135 140 Thr Trp
Ser Glu Cys Thr Glu Pro Lys His Ile Val Glu Trp Asn Val 145 150 155
160 Ser Leu Phe Ser Ile Leu Leu Ala Leu Gly Gly Ile Glu Phe Ile Leu
165 170 175 Cys Leu Ile Gln Val Ile Asn Gly Val Leu Gly Gly Ile Cys
Gly Phe 180 185 190 Cys Cys Ser His Gln Gln Gln Tyr Asp Cys 195 200
42 145 PRT Rattus sp. 42 Met Ser Ser Phe Ser Thr Gln Thr Pro Tyr
Pro Asn Leu Ala Val Pro 1 5 10 15 Phe Phe Thr Ser Ile Pro Asn Gly
Leu Tyr Pro Ser Lys Ser Ile Val 20 25 30 Ile Ser Gly Val Val Leu
Ser Asp Ala Lys Arg Phe Gln Ile Asn Leu 35 40 45 Arg Cys Gly Gly
Asp Ile Ala Phe His Leu Asn Pro Arg Phe Asp Glu 50 55 60 Asn Ala
Val Val Arg Asn Thr Gln Ile Asn Asn Ser Trp Gly Pro Glu 65 70 75 80
Glu Arg Ser Leu Pro Gly Ser Met Pro Phe Ser Arg Gly Gln Arg Phe 85
90 95 Ser Val Trp Ile Leu Cys Glu Gly His Cys Phe Lys Val Ala Val
Asp 100 105 110 Gly Gln His Ile Cys Glu Tyr Ser His Arg Leu Met Asn
Leu Pro Asp 115 120 125 Ile Asn Thr Leu Glu Val Ala Gly Asp Ile Gln
Leu Thr His Val Glu 130 135 140 Thr 145 43 318 PRT Homo sapiens 43
Met Met Leu Ser Leu Asn Asn Leu Gln Asn Ile Ile Tyr Asn Pro Val 1 5
10 15 Ile Pro Phe Val Gly Thr Ile Pro Asp Gln Leu Asp Pro Gly Thr
Leu 20 25 30 Ile Val Ile Arg Gly His Val Pro Ser Asp Ala Asp Arg
Phe Gln Val 35 40 45 Asp Leu Gln Asn Gly Ser Ser Met Lys Pro Arg
Ala Asp Val Ala Phe 50 55 60 His Phe Asn Pro Arg Phe Lys Arg Ala
Gly Cys Ile Val Cys Asn Thr 65 70 75 80 Leu Ile Asn Glu Lys Trp Gly
Arg Glu Glu Ile Thr Tyr Asp Thr Pro 85 90 95 Phe Gln Lys Glu Lys
Lys Ser Phe Glu Ile Val Ile Met Val Leu Lys 100 105 110 Ala Lys Phe
Gln Val Ala Val Asn Gly Lys His Thr Leu Leu Tyr Gly 115 120 125 His
Arg Ile Gly Pro Glu Lys Ile Asp Thr Leu Gly Ile Tyr Gly Lys 130 135
140 Val Asn Ile His Ser Ile Gly Phe Ser Phe Ser Ser Asp Leu Gln Ser
145 150 155 160 Thr Gln Ala Ser Ser Leu Glu Leu Thr Glu Ile Ser Arg
Glu Asn Val 165 170 175 Pro Lys Ser Gly Thr Pro Gln Leu Arg Leu Pro
Phe Ala Ala Arg Leu 180 185 190 Asn Thr Pro Met Gly Pro Gly Arg Thr
Val Val Val Lys Gly Glu Val 195 200 205 Asn Ala Asn Ala Lys Ser Phe
Asn Val Asp Leu Leu Ala Gly Lys Ser 210 215 220 Lys Asp Ile Ala Leu
His Leu Asn Pro Arg Leu Asn Ile Lys Ala Phe 225 230 235 240 Val Arg
Asn Ser Phe Leu Gln Glu Ser Trp Gly Glu Glu Glu Arg Asn 245 250 255
Ile Thr Ser Phe Pro Phe Ser Pro Gly Met Tyr Phe Glu Met Ile Ile 260
265 270 Tyr Cys Asp Val Arg Glu Phe Lys Val Ala Val Asn Gly Val His
Ser 275 280 285 Leu Glu Tyr Lys His Arg Phe Lys Glu Leu Ser Ser Ile
Asp Thr Leu 290 295 300 Glu Ile Asn Gly Asp Ile His Leu Leu Glu Val
Arg Ser Trp 305 310 315 44 128 PRT Homo sapiens 44 Met Arg Thr Ala
Leu Leu Leu Leu Ala Ala Leu Ala Val Ala Thr Gly 1 5 10 15 Pro Ala
Leu Thr Leu Arg Cys His Val Cys Thr Ser Ser Ser Asn Cys 20 25 30
Lys His Ser Val Val Cys Pro Ala Ser Ser Arg Phe Cys Lys Thr Thr 35
40 45 Asn Thr Val Glu Pro Leu Arg Gly Asn Leu Val Lys Lys Asp Cys
Ala 50 55 60 Glu Ser Cys Thr Pro Ser Tyr Thr Leu Gln Gly Gln Val
Ser Ser Gly 65 70 75 80 Thr Ser Ser Thr Gln Cys Cys Gln Glu Asp Leu
Cys Asn Glu Lys Leu 85 90 95 His Asn Ala Ala Pro Thr Arg Thr Ala
Leu Ala His Ser Ala Leu Ser 100 105 110 Leu Gly Leu Ala Leu Ser Leu
Leu Ala Val Ile Leu Ala Pro Ser Leu 115 120 125 45 299 PRT Mus sp.
45 Met Ala Gln Asn Leu Lys Asp Leu Ala Gly Arg Leu Pro Ala Gly Pro
1 5 10 15 Arg Gly Met Gly Thr Ala Leu Lys Leu Leu Leu Gly Ala Gly
Ala Val 20 25 30 Ala Tyr Gly Val Arg Glu Ser Val Phe Thr Val Glu
Gly Gly His Arg 35 40 45 Ala Ile Phe Phe Asn Arg Ile Gly Gly Val
Gln Gln Asp Thr Ile Leu 50 55 60 Ala Glu Gly Leu His Phe Arg Ile
Pro Trp Phe Gln Tyr Pro Ile Ile 65 70 75 80 Tyr Asp Ile Arg Ala Arg
Pro Arg Lys Ile Ser Ser Pro Thr Gly Ser 85 90 95 Lys Asp Leu Gln
Met Val Asn Ile Ser Leu Arg Val Leu Ser Arg Pro 100 105 110 Asn Ala
Gln Glu Leu Pro Ser Met Tyr Gln Arg Leu Gly Leu Asp Tyr 115 120 125
Glu Glu Arg Val Leu Pro Ser Ile Val Asn Glu Val Leu Lys Ser Val 130
135 140 Val Ala Lys Phe Asn Ala Ser Gln Leu Ile Thr Gln Arg Ala Gln
Val 145 150 155 160 Ser Leu Leu Ile Arg Arg Glu Leu Thr Glu Arg Ala
Lys Asp Phe Ser 165 170 175 Leu Ile Leu Asp Asp Val Ala Ile Thr Glu
Leu Ser Phe Ser Arg Glu 180 185 190 Tyr Thr Ala Ala Val Glu Ala Lys
Gln Val Ala Gln Gln Glu Ala Gln 195 200 205 Arg Ala Gln Phe Leu Val
Glu Lys Ala Lys Gln Glu Gln Arg Gln Lys 210 215 220 Ile Val Gln Ala
Glu Gly Glu Ala Glu Ala Ala Lys Met Leu Gly Glu 225 230 235 240 Ala
Leu Ser Lys Asn Pro Gly Tyr Ile Lys Leu Arg Lys Ile Arg Ala 245 250
255 Ala Gln Asn Ile Ser Lys Thr Ile Ala Thr Ser Gln Asn Arg Ile Tyr
260 265 270 Leu Thr Ala Asp Asn Leu Val Leu Asn Leu Gln Asp Glu Ser
Phe Thr 275 280
285 Arg Gly Ser Asp Ser Leu Ile Lys Gly Lys Lys 290 295 46 80 PRT
Homo sapiens 46 Met Asp Cys Val Ile Thr Gly Arg Pro Cys Cys Ile Gly
Thr Lys Gly 1 5 10 15 Arg Cys Glu Ile Thr Ser Arg Glu Tyr Cys Asp
Phe Met Arg Gly Tyr 20 25 30 Phe His Glu Glu Ala Thr Leu Cys Ser
Gln Val His Cys Met Asp Asp 35 40 45 Val Cys Gly Leu Leu Pro Phe
Leu Asn Pro Glu Val Pro Asp Gln Phe 50 55 60 Tyr Arg Leu Trp Leu
Ser Leu Phe Leu His Ala Gly Ile Leu His Cys 65 70 75 80 47 1497 DNA
Homo sapiens 47 tttttttttt tttttttttt cacgttacaa tctcactata
ttcggaatct aaagtagtca 60 tccttagaga agtacagagg agaatggtgg
gtacagaggc cagggcgggg agtgtggacc 120 gattggatgg ggaaagggag
agtttggtca tcaggcatgc attttcagtt agacaagagg 180 aataagttct
gatgtcttgt tgcacagcat tgagggctgc taattctact ggaatcagcc 240
tcagactcct ttgctggagg ggtcgtgaac cctcaaacaa gcccgagcct ctgcattttc
300 ttacttgggg atcttgatct ctgagtactc attgttggtg gcttcttgac
ctggataggt 360 cctgaggctc ccccttatga aagctgaggg gtgcatactg
ggatctctct ttcctcccct 420 gagggagtgg gcagccaggc catggtgtcg
ggggttatca tctgcccagg actcatccag 480 ttaccctgag aggctgagcc
cctgaatggt gtttgcatcc ttcatgccta tgtctcccac 540 gtccgctgct
ggtcttgccg atttcttcct gcaggacctc actacaatga agatgacaca 600
gaaggagagg aagaccaggg ctgtggctcc agttcccccg accgccccca gcaacactcc
660 tgatacaggc ttcattttgc ctgtgtactc ctgttgcagg gagaggttca
gggaaacgtg 720 ctgggaaccc agagagttct gagctcgaca ggtgaattcc
ccttcatccc ccaggtgcac 780 ttgcagctcc agtaccagag ggtttgaggg
ctgtgagggg tacagggtca gactcctcca 840 ggtccagctc agcctggcag
ggggattgct gtcaacagca cagaccaagc gcagagactg 900 gccctctagg
actgaaagag atgagctgtt ccccagagct gtggatgctg tgccttctcc 960
ttggaagaca gtcacagtca agttctgagg agggtatgtc acgttcacag agagctggtc
1020 atatttataa ttccatttta tatttccttt ctccatacga aagaagtatc
tccccgcatc 1080 actcattctg gcatctctga tgctcagggt gcaatttttg
gtctgtgggt ccccaaggag 1140 gtggaatcgg tcccgagttt cctcctgcac
tgcccaagct gggttgtttg tggccactgg 1200 agccttccag cttatatcat
tccctgcccg gaaccagtag ccatgaactg ggtcagagtc 1260 agtctggctg
tccactgggt aggagaagga gcagcgcaca tggacacaca tgccctcttg 1320
cacggtcacg gaactctgca tcgtcagcga gtaatccttc cggttactct tctgtccttc
1380 caccctctcc ctcccccaga gcaggggcag cagcagcagc agcagcatat
ctggggttgg 1440 aggtgccagg gccgcgaggg aacgtctgtt cctcagggtt
cttctctcag gaactgc 1497 48 1849 DNA Homo sapiens 48 ttttttttta
caatcaaaat gcacatttat taaactgtac tgtgtgaggg aatgtctgac 60
caaagagcat tcccacaggc actcggggca ctggctggaa agcgctctca ggtgttcttc
120 tcagaaccgg agaagctgca catcacttgt atttgtaaaa tccttctcca
gtcttcttgc 180 cgaacttgtt ctctgctacc agcttattta aggatgggct
gggctgatgt aatgggttct 240 ctgcatccat ttcatgccac ccatccacga
tgaacttcgt agtatccagt ccgacataat 300 ctagaagctc aaatgggccc
atggggtaac cggctcctaa tttcatagca gtgtcaatgt 360 cttctttgga
tgcgtcacct tttaatcaaa agcagagatc caggatatca atctgggccg 420
gccatgcaat tgctctggct gggactgtag tgggggtggt gggagttcca gaaacctttc
480 tcagaagccc acagggaaag ccaggaggcc ctcgttcata cagcctgatt
gcttccatga 540 ggtatggaac caggaggcgg ttcacaataa acccaggagt
gtccttgcaa gaaacaggat 600 gctttcctag ggttttgcta aagtctacca
aagattcaaa tgtcttctgg ctggtcattg 660 gtgttttaat gacctccaca
agtttcatga caggcactgg gttgaagaaa tggaggccag 720 cgaatcggtc
ttgtctggtg gtggcattag ctatgcttgt aatctgcaag gaggaagtgt 780
tgctggcaaa gattgtatgt cttggtgatg tggcaggtgg ggctgcatgg agattcaggg
840 acagctcctc tcaggcagac cctcaggaac cttaaaagga tgcaggaagc
ccagcgtggt 900 ggctcacgct ggtaaaccca gcactttggg aggcctaggc
ggggggatcg ctttaggcca 960 gcagttcaag accagcctgc gcaacatagc
gagaccccca cctctgtatc actgcagctc 1020 ccagagagct cccactcagg
gtgaagctct gagtcttggt gatttttagg agtttatgga 1080 aacagccctt
ctggtggatg acgtttggag acacatcgcg tccgtcacag gacacacttc 1140
cgatggattt acagcaactc ctggggtagg tgtggcccgt tgtcatttcg aaggaagagc
1200 cagaaaaatc tgtgtagtta ttcaccccac agcactttag cttctccatg
accaagttcc 1260 actgtgtaga atagtcgtct ggctcgttgt aacctctgta
attcttcctc agggtcacga 1320 aggtgtgttc caaggccaca tctccaacaa
ttggaaagaa aagaaggacc actgtggcag 1380 ctgtaacttc catgatgagg
acaataacca ttgacaggat gcaaaacaag agcgtgcctc 1440 tgctctcttt
agtcgctcca taccacccgg cacagccaag cagtaccgtg atgcatccca 1500
tcaccaggca caggttgcca acgtgaagga ggtatgcgga ggacagcccg aggacattcg
1560 tcagagaggc ccctccacat ttaccaccaa tgcccaggcc aactaggatg
atgccagaca 1620 cagccacgaa gccattgagt aaagataaca gtttcttcaa
ggaagaatac ggagtgtgga 1680 tttcagccat gctgaacaga ctggggcacc
ccgaacattt aagttaacat cttcctgaac 1740 ctctgggcta ctgtttccca
agatgatagg gatctgagga aggggcgcca atgctgactt 1800 gctctgcccc
tctgtgtcct ggctccctga tggttgtcca ctcgtgccg 1849 49 741 DNA Homo
sapiens 49 gctttaggcc agcagttcaa gaccagcctg cgcaacatag cgagaccccc
acctctgtat 60 cactgcagct cccagagagc tcccactcag ggtgaagctc
tgagtcttgg tgatttttag 120 gagtttatgg aaacagccct tctggtggat
gacgtttgga gacacatcgc gtccgtcaca 180 ggacacactt ccgatggatt
tacagcaact cctggggtag gtgtggcccg ttgtcatttc 240 gaaggaagag
ccagaaaaat ctgtgtagtt attcacccca cagcacttta gcttctccat 300
gaccaagttc cactgtgtag aatagtcgtc tggctcgttg taacctctgt aattcttcct
360 cagggtcacg aaggtgtgtt ccaaggccac atctccaaca attggaaaga
aaagaaggac 420 cactgtggca gctgtaactt ccatgatgag gacaataacc
attgacagga tgcaaaacaa 480 gagcgtgcct ctgctctctt tagtcgctcc
ataccacccg gcacagccaa gcagtaccgt 540 gatgcatccc atcaccaggc
acaggttgcc aacgtgaagg aggtatgcgg aggacagccc 600 gaggacattc
gtcagagagg cccctccaca tttaccacca atgcccaggc caactaggat 660
gatgccagac acagccacga agccattgag taaagataac agtttcttca aggaagaata
720 cggagtgtgg atttcagcca t 741 50 1288 DNA Homo sapiens 50
tttttttttt ttttttttta acattgtaac aggtttatgc attttgaagt gccttctaca
60 catccaccca gaggctctgc tgatttcact tatgcccagg ctataaaatg
cctttctctc 120 atcccccagt agagcactgg gatcaccact aggcctaggg
ggcatatcaa gggtttaata 180 gactggggga atgggcaaca gaactggcta
ccttagaggc tctggaatgc cccccaccca 240 tccacccacc aatggaagga
aagtcaggca tcgcctaaaa ggagtggtcc ctatctagcc 300 ccaagtctgg
agcagaaagg gcaggtccat tctggcccaa gtgacattgt tagatcctgt 360
cccctccccc aatcactgct gcttgccagg gtgcctcttc acagttccca tgtggcagca
420 gtagtggcag aggcagaagt ggacttattg tagattgcag tacagataca
tggacacaat 480 tcatggcagc cagctcgagg cccccaattc cagctgccac
accacccacg gtgactgcat 540 tagttcggat gtcatacaaa agctgattga
agcaaccctc tactttttgg tcgtgagcct 600 tttgcttggt gcaggtttca
ttggctgtgt tggtgacgtt gtcattgcaa cagaatgggg 660 gaaaggcact
gttctctttg aagtagggtg agtcctcaaa atccgtatag ttggtgaagc 720
cacagcactt gagccctttc atggtggtgt tccacacttg agtgaagtct tcctgggaac
780 cataatcttt cttgatggca ggcactacca gcaacgtcag gaagtgctca
gccattgtgg 840 tgtacaccaa ggcgaccaca gcagctgcaa cctcagcaat
gaagatgagg aggaggatga 900 agaagaacgt cacgagggca cacttgctct
cagtcttagc accatagcag cccaggaaac 960 caagagcaaa gaccacaacg
ccggctgcga tgaggaagta gcccacgttg acaaactgca 1020 tggcactgga
cgacagtggc ccgaagatct tcagaaagga tgccccatcg attgacaccc 1080
agatgcccac tgccaacagg gctgcaccac acagaaagat gagcaaattg aagaggatca
1140 tcatggtctt aatgaagctg aagcactgca tggtggctcc tgttcagggc
tcttggcagt 1200 gagttctgaa agagggaact gctgaggctc cacaaaggac
aaaacagctc ccgggtgttg 1260 ccactgagtg ggcaggcaga gggacgcc 1288 51
1236 DNA Homo sapiens 51 tttttttttt ttttttttga atgctcagta
tctatttatt aaatgaatgc acaattgaaa 60 gagtgaagat tgaatgtcta
gatatttgtt cctctaaccc acaagctcac atgcagcaga 120 gcacacgaat
gggtgtgcca aggccccaga ggctggatgc ccacatatgt gctgatgcat 180
gtgccctagg gagtccctct tggttaacat ctgtttaaaa tctctgtctt cttcttgata
240 gagaaacagc atggtttcct agactgaatt gaagaatgaa ggtgagagta
caaacagtgg 300 gagagacgtt tcctcagctg tcagaacagg aacgacctgg
gttatggaag cccagaaagg 360 gaggaggact tcttttggtc ccagtgaaag
atgcttccag aatctgtagc cttacttatt 420 tgcttggatc tcactggaat
aacttggtgg tgaggtcacc ggttctgggg tgatcactgg 480 gtttgctgca
tagatgtttg gatagatgac actcacattg cttgattgac agcagaccaa 540
ctggcagcca aagtgggaag atgcgcatgc gatgccaaac tccaggaggc agaagaccag
600 cagcacgcca gaaatcgcca ttccagggtt cacaccccag gcgtaaggat
aatagtcggg 660 gtaggcatat gggtggggaa tacttagatc tgtgatgaag
agtatgactc caactgcaga 720 gcagattgca ctgacgatgt tcaagcccaa
actgccagac agcaggcaat aagaatatgg 780 ctgattttct gctgccacgg
agagagatcc tgaaatgata aaccacaagc ctccccagaa 840 gggaaagcct
ccgtagaatg aaatagacag gtattcccct acgagaaccg tcgccatgat 900
ggagccgagg ccgatgtgag ccaggccaat gatgatctgg atggccccca aggttttgcc
960 ttctttcaga gctttctgca caggctgccc attcacattc gacaccaaac
taggtgggtt 1020 cccaggaact aggtggactt gcggctggct gtttggatac
aggggcacgt gagacataat 1080 tcctggggtc acaggataac cattgtgggg
tgccaccacc aacacagaat tggccaccgg 1140 aactgctgaa gtcatcgaat
tcatgcttgc tctgccagca gccacgtatc tcttgatcct 1200 attccttttc
tttgctgtgg ggaccttgtt tttttt 1236 52 1115 DNA Homo sapiens 52
tttttttttt tttttttttt tacttcattt acaatttact agctcttcca gtgtttcaga
60 gggatacagg gtttcaacga tctaacatga atgggataga aggtggactt
agaacatagc 120 aaacatacat cttgattgaa tcagcccact gcgagcacgg
atcttgattg aatcagccta 180 ttggtgtagt tttaggtcta catacatctt
gattgaatca acctactggt gtagtttttt 240 ggtctatcag taagtagtgt
tgtcagttct ccagccaagc aactttctaa ttcacagggg 300 ggaccctaaa
tgtccttaaa ggtagagagg aataggccca cagatacatt gggttacact 360
atctcatact ggttatttgt tatggcacga gagaggcagt aggcgagaaa gattccaatc
420 agttggaagc aagcaactcc aaaggaaatt cctgcaacga ctcccatttc
tgactctata 480 atggtcatca cctttataaa acaaccttca ttgtttactt
tgtctgcatc tctctgtgga 540 gtacaatctt caagtttaca gcaactctta
ggaaatcctt tttctgagta ataattagta 600 tctgtccaat ctctataatc
ggtgacacca caacaatgca acgtattttg gatcttgtct 660 actgcatggc
ttctataatc tcctgtagag ttatactgct tcaaagcctt ctcataatta 720
ttcttaaagc tgttcttaat ctcatgtctg aaaacaaatc ctacgatggc agcgaccagt
780 tcgaccaaaa aaacgagagt cagaaacatt gcatacagtt ttagcatcca
tgcagaagct 840 cggcaggtag caaaacaacc aaaggtgccc aaaagaataa
tgacggtacc agtagcaatg 900 agcacgaagg ggacattggt ggccttctca
tttaaaagag aaaagtaatt ctccaggctc 960 accttgcccc aaatgccaac
tgcaagaagg ataacgccag tgatccagaa aataaaagtg 1020 tagattagca
gaacgctctt gaaacaagta atgactggtt tagtctgcag tctccgagac 1080
ggggacgcca tgactagccc gagaccctgc tcgtg 1115 53 1662 DNA Homo
sapiens 53 tttttttttt tttttttttg ttgggggctg atacagagtt tattgaatta
gatttttcta 60 tttacaactg agatcacatc tacatactat tttgctagtc
tacatgggta cattatttcc 120 aacaagctta agacttacca tgaatgggct
cattcataca aaaacacact cacactaatt 180 cttttaaaac agtagtgcat
acattatact cctcctataa agccaacttt gattaaaaac 240 cactagtttc
aaagctcagt ctctgatttt gaagatgaac caagatatac gccatatgat 300
cctacaatct attttagtca ttttgtacag ctgctatctt attggactac agtaaatatt
360 ttttaaaagg acaccaatga ggggcaccat ctggtgttaa ccttaaccag
aaagctggtt 420 tcctcctcct ccccccaaaa acctttggcc aagagttctc
cactgtgaag actgaaagga 480 cctggtgaca tttcggcatc agtcctgtta
ccacttggag gtaacagaag caggctcgtg 540 tcctccttta attctaccac
actacatgac tcgcaattgg ttctgaaatt agaacgttca 600 ccatcgtact
taaaatctta ggggcatgaa gagtcagcta gaacaaggaa aaagaaagtc 660
gcaggtagta ggtaagtagg tgggcacatg aaaagccaag ctgctctgtc caacaccagt
720 gtacatgtgc tttaactaaa tgaactccag aggccaacag cagcagacct
gctcaattca 780 ccttccaaat cagaacaaga ccaaaaagct caggcttgag
attgtcaact atgcataggt 840 tccgccagtg atgaagagct cgtaagcagg
atctctactc cttctgcaca acacgatgca 900 agcacacagc atgcccagca
gctgaatagc tgcaaatgcc agtgcggccc agatcacatg 960 catcatgatt
tcttgtagct tcttcacaac tagagcctca cacccctcag catagaggtc 1020
ggaagggtgg gccaggctgc cattacaatt gctggcagtc tctctgcagc agctaagagg
1080 gacactctgg tttttggttt ctttgaacca atctgtattt tcccagtctg
agtagttgtg 1140 aattccacaa caatgcagct gtctctgtac ataatcaata
gcccggctag cagcatcagg 1200 gttggttcca ttgtaggtct tatacacttt
ctgaatgctg cgatcaacct cattttccac 1260 ctttgctctg taaacatatc
ccaaaaccac tacaacaact tctgtgacaa aaaccaagag 1320 caggatgatg
acaaacgtgg caagtccaca gcgactttcc cggattgtgg cacagcagcc 1380
aattagccca atgatgaaaa gcagggctcc tacagctatg atcactacag cagggatgag
1440 cgtgtacaca tcttcaaaga agtggtcata gtcatcataa gtgatgaaga
cataggctcc 1500 cacatagcat aaaatgccag ctgcccccca gaagatgagg
ttgagaaaga ccagcacggt 1560 cttggaggag gtgatgccgc actggcccat
ggcgccggtg gcccgcgaag gcccggcccg 1620 gagagcgggg ctgcgctcac
cgagagagcg gcaatgctcg tg 1662 54 1345 DNA Homo sapiens 54
tttttttttt tttttttttt tgctgctcac actttattaa gatgcaccag gagccccacg
60 ggcccactat ggaatgtagg tgaggggtcc acggccccgc ctggagcacc
aggaccagtg 120 gggccacctc caggatgcca agacccggct cctccagcgc
caacctgttt tccaggaagc 180 tgggggccac aggctggctc ccgtgtgaac
actgctttgg agaatacgta aatataaaaa 240 gtgctggagg cccctcccac
ccctgccctg ggttccgcag ccagcacgtg gccacccctc 300 ccaggggggg
tccggaggcc ctgaagccac ctgagctagg gccttccaga aacagggttc 360
cgggggctcc aggcacctgt ccctccctcc ctcctcccag catggggcag aacaccgaag
420 ctggtggtgg gaaaagcagc tgtgggtgcg gccatctccc cgtgggcgtc
cttttggcag 480 agaagccggc ggtgggcggc ctacgcgcag taggtgtctg
ccttgaccac ttggcagtac 540 atggtcatgg cgaaggtcag gcccaggatc
tgcaccagcg ccgtgcacag cccaaagatg 600 cccacagcca gcaggttctc
ctgaagccac accttcaccg tctcgtagca cggcgccttc 660 caccaggtgc
cgggggcgtg cagcccacag ctctcactga actccaagca gcaggagtca 720
ggtacccgcg tggcgttgta cacctcgaac cagtcagtgt agttggagac gccacagcag
780 cggaagtcgg tctggatgat gctccaggcg ttggtgaggc ccacgttgcc
ctgcgtgccg 840 tacaggtgca agcctttctt caggtcttgc tgggcatacc
tgtcaatctt gtccgtgtag 900 gcgaagaaga ggatggcgat ggtggcctcc
agcaggaaca ccagcagcag cagcaggaag 960 aaagtgagca ggaggcactt
gttctccttg atggcaccca ggcagcccac gaagccgatg 1020 gccatgacaa
aggcgccggt gatgatgagc aggttggcag ccgacaggga cgggaaggaa 1080
gaagacagcg tggcgaagct cccctgtgtg gcggccagcc agatgccgac acccagcacg
1140 ccacagcctc ccagccagaa gaacaggttg aaggcgaaca tgaggtactt
gacggcctgg 1200 aggcaggcgc gcgccatgcc gcagcgcttc agttctgggc
tggccacagg aaagagacca 1260 ggcggtgctc agggtccctg aaggctgacc
agtgggcagg caggtggtgg gtgcgaccaa 1320 ggaagcccca agctctgcgc tcgtg
1345 55 734 DNA Homo sapiens 55 gaacatttgg tatgaaagct ttagattgca
attttcatgt agaagtagct tcatatcaca 60 tctcgtgagt ttcgtatcgc
acagcagagg accatgctga atatcatgcc aaagatcgtc 120 agacctgcaa
ttccaatacc gacaattcca atgagctgga gcttaacact gattatggtc 180
tcaatttcat cgatgcaatt cttgtgtcct agaagctcct ttgggcatgt aggttggacc
240 tgttcggagc tttcttttcc acagcactga aatgttgagt ggaaggtgat
gagtgtccca 300 ttgccttttc ccctgtcttt aaggtaatca ttgtaagcct
cttcatacat ggtctgaaca 360 tgtcggatag ctaccccctt gcctataaaa
gcaaatactc cagtggttac ttcagcagca 420 aatatcacca ggaggcaggt
aaaaaatgat ccaagcacac attgcgactc ccgcatggct 480 ccgcagcatc
cgaagaaccc cacggccatc atcagggccc cggctccaac cagaacatac 540
agccccacat agaaatactc tggggacttg tcctctgatg ataactcctt tatggcacct
600 ccgaaccgaa accatagtcc aaaagcaatg acggccgatc cagccagcca
gaagagcagg 660 ttgaagccaa gcagcaggta cttgatgcac cgcaggcccc
cgcggaagcg ccccatgctg 720 cggcccggcg gcgc 734 56 577 DNA Homo
sapiens 56 tttttttttt tttttttttt taaacattga aaattttatt ttcacagggg
atttgcataa 60 aaagaacatt atttttgttc tgtgtatata taagtatttt
tgtttcctta acttgtttct 120 gttgcccaca cacaactagg agaagatgct
tttctttatt ttggtttggc caaagatgct 180 aatggttaaa ttatgaagga
ctttgtttta cttatgttaa gtggtgaaaa ctgtagttct 240 taatctatga
agaattctct aggtggctat acaagaaaaa tacaaaaagt taggaaaaca 300
tgtaaacgta agtatgaggt atttcataga tacagtgccc atacaaattc tctttcccac
360 aattttcaac tgccagatct cttgctttag tcttttttcc ttatatttgg
agaaacagaa 420 gagtttgaca taaaagtccc tttgagggat gtgagggttg
cagtagttta cagcagggtc 480 agaaaatgaa agtaataaag catatttaca
tgttttgtat aggaccaaaa tatttcccct 540 aaaaaggtgt taaaagtttt
ttagtcccat aaacact 577 57 936 DNA Homo sapiens 57 58 738 DNA Homo
sapiens 58 gttttttttt tttttttttg agagcgcaaa gcagtttatt ctagcgagca
agggagtgag 60 cgtccaggaa ggagcaggtg taacccggcg gtcagtggag
cctcagtgag gtgtgtcctg 120 ttttttcctg caatcgccgc agaagacacc
aatggtcgcg ttcaccagct ggatcccaca 180 cagtactatc tccaggcagg
aggcggccac cagcagcgag aagagcgtca cattccaggg 240 gaccacgcga
gggggcgcct cgcaccgatc ccatagagtg cggttgagca agtaagctcc 300
cgcggtgtct tcgaagtggt agccccactc gccgttcatt aagcatctgg gtccatttcg
360 gagcccagct ccagacaccg agaggcagta gatggcacca agcaccccga
acgccgagga 420 gaagaccgag cgcagcatcc tgcagcggtt tccacagcac
ccagcaccac agcagccctt 480 gccccctgcc cgaacggctg caatccctgg
acacagtacc attaggcccc cgccaatgaa 540 gccgcccatg agccagactt
gcaagctgag atggttggtg ttggtccagg aggtctcccc 600 attaggtacc
agcaggaggg cgttggccac aatgcagacg aggcagaggg taatgaggga 660
gagccccaca cagcgggcac attttcccgt acacatggtg aggtgtcagg aaggacaggc
720 ggtgagtgaa agtaagct 738 59 1071 DNA Homo sapiens 59 tttgagttag
aattttgggg tgtttccaat tttccagaat attaaataat gttgcaatat 60
acatattgca taaaggtttc tctcatattc ttatttactt ccctagtaca gattcttgga
120 aatggatttg tttgatcaaa gggcagaagt atttctgaag tttctgatgc
atattactta 180 aaaaacatac taaatggaag ggtggtatac ttgcatgtgc
agtgaggact gcaaattttt 240 tacaaataaa caccaaatgc agaaagcacc
atcattttca atattgccct caagtctaac 300 acagttgata taatatttag
atagatggcc atgtcttgat aatggaaaac attttgtcct 360 tattcaaatg
attccaggct ggaagatcac tgaatagctt ccacacagta tcttggatag 420
ttgcatgact actctgatga ggcagatgat cacttgaagc ccactgaggg ttatgagaat
480 ggaaaataaa atgatgttcc actccacaac atgtgcaggt tccaggcact
gaatccatat 540 gctagaatct gtaaggaaac gtccagcagt gccttcaaaa
gcatactccc agccatcaag 600 ggtgcggcaa tatggccctt ggacaagacc
caaggcagag atgaccaggc agtatccaga 660 aaaagcaatt ccgagggaag
aaaagataat tgacagcagt gtcacatatt ttttgctgca 720 gttttcactc
tggcaacatt tatagttgtt attattctcc agtaccagaa gaactgttgt 780
tactataagc atcatgatgc ctgagaaaca gattccttca aaataccaca cgtagttggt
840 gagtttattg ctggatgcat aggaagtttg cccattcggg aaatacaata
atatgttcac 900 gattatactc caaagtgcaa gcggaatcag caaacaactt
aggcagcctc cacacttccg 960 agaccccatt ttgccctgct tagaaccact
tccagggtca gagcccttca cttcatattc 1020
atgaggagac ggggaattgg aatatacccg cagccgacaa tctctcgtgc c 1071 60
865 DNA Homo sapiens 60 tttttttttt tttttttttt tttttttttt tttttaaata
aataacctta tttatttatt 60 tgaccacaaa taaataaata ataaagaata
aatgaccaca ttctttaagc cattgtattt 120 tggagtatct ttgttagtct
acaacacttg cctctgtgag tgcagtccag gccctgaagg 180 cagatggagc
catatcccag gctcctggtg gaggaaggtg cagagttcga ggaacctttg 240
cactcttgtg ccttccctca ggcccaaagc tcctgtagac tcagtctcgt gaccccagag
300 gtgaaccagg ccctgatgtg ctggtgttca ggtggatggt ttaatgaggg
gaggagagtg 360 gggccctggg gagtggctga ctcttgtttt ctgcggtatt
tcctggaacc atccttcagg 420 agtggacaca gtagagctgg acacttccac
tgatccggag ctcccgcagc tgctccaggg 480 cctgctggtt catgctggtg
gcccccagcc cctgcccatt gagcgccagc ttcagccctc 540 cctcctggaa
caggagcagc acctcaaaga atctctgggg gtaaaagagg aagggggctg 600
agatcagttt cttctgcccc cagcgggaga tccaggccag agttctgtct gcgaaggagg
660 ccctgagtgt cacaggagca tgggcagcct ggtccctcag gctcacagta
aaatgcttcg 720 gctcttgcaa gaccagtccc cgtactatga tgacctgccc
aggcgagaga ccctggggaa 780 gagcatgtga gcagggcacc tccagcctgg
ggctcatcag caggaaagga tgtccagctg 840 ggtactctct gctgccctcc acaaa
865 61 441 DNA Homo sapiens 61 tttttttttt tttttttttt tactggttta
aatcatttat ctccatgtag agatttgagt 60 acaaaaataa acggcaacaa
aacagaagga gtgtgaaatc cggggatcca cagggcttct 120 gtcctccacc
ttccatgcag ctgggggctg catcctctgt ggggtggctt catcctctgt 180
ggggtctgtg gggcctgctc caagtcatca gcattccatg cccacctgga cctggtccca
240 gactttcggg gaagccctca gaggctccac tgtgttcgtg gtcttgcaga
agcgagagct 300 ggccgggcag accacagaat gcttgcagtt gctggagctg
gtgcacacgt ggcagcgcag 360 ggtaagggct ggccctgtag ccacagccag
ggctgcaagg agcagcaatg ctgtcctcat 420 ctctgatgtc gtctctcgtg c 441 62
1066 DNA Homo sapiens 62 tcaaaaactg gagaagcaga tccacttctt
gtgggggtgg agttcttggt gactaggctc 60 atttcttacc cttgatgagg
ctgtcacttc ccctggtgaa actttcatcc tgtaggttca 120 gcacaaggtt
gtcagctgtg agatagatac gattctgtga tgtggcgatc gtcttggaga 180
tattctgggc tgctcgaatc ttgcgaagtt tgatgtagcc agggttcttg ctcagtgctt
240 ctccaagcat cttggcagcc tcggcctcac cctcggcctg cacaattttc
tgccgctgtt 300 cctgctttgc tttttctacc aagaattggg cccgctgggc
ctcctgctgg gccacttgtt 360 tggcttctac agcagctgtg tactctcggc
taaagctcag ctctgtgatg gccacatcat 420 ccaggatgag gctgaagtcc
ttggccctct ctgtcagctc ccggcggatc aacagggata 480 cctgggcccg
ctgggtgatc agctgtgagg cattgaactt ggccaccaca ctcttgagca 540
cctcgttgac aatggacggc aacactcgtt cctcgtagtc cagccctagg cgctggtaca
600 tgctaggaag ctcctgagca ttgggtcgag acaacactcg cagggagata
ttcaccatct 660 gtaggtcttt ggagcctgta ggggaggaga tttttcgagg
tctggcccga atgtcataga 720 taatggggta ctggaaccaa gggatcctga
agtgaaggcc ctcggccagg atagtgtcct 780 gctgcactcc accgatccga
ttgaagaaga tggctctgtg cccgccttcc acggtgaaca 840 cagattcgcg
cacaccgtag gccacggcgc cggcccccag caacagcttc agggccgtgc 900
ccatgccccg gggcccggcg ggcagccgtc ccgccaagtc cttcaagttc tgggccatgt
960 ctgatcttga ggccggcggc actggaggtc agaagggggt gccggcccgc
ctctaccccg 1020 ctccggctta ggtactgcac ccttcacacg agggttcggg cccgct
1066 63 704 DNA Homo sapiens 63 gcgatagggt gccagccgga ccaccttcct
ccaggtcccc ttcagggatg tcatttgaaa 60 agacccacaa gacaccgagg
gcagtgcacc cacactgggg gagggacaca ccaggcagcg 120 gccttcatcc
agacacttca ggagcagtag tcttcggcag gttggaccga ccataccctg 180
gcccaggaac ctttccccag gggtccgcct gggtgtccat cacaggggtc gggttcgggg
240 gcattgagga cctcttaccc aacatgtagg aaaagagaca gccagacctg
taaaactgat 300 ctgggacctc agggttgagg aaggcagcag cccaacacac
cttgtccaaa acagtgcacc 360 tgggaatggg catatggtgc tcagcgcccc
agaagcagcc tgagtctgag gctccgacat 420 gggagggagt gcgtgagccc
cggccccagc aggctgccat gcccagcgtc ctctcctcca 480 ctactccagg
cctgcctcgc ctcacctggg agcagagtgt tgcttcctca tggaaatagc 540
cgtgcatgaa ctcacagtat tcccgggtgg tgatctcaca gctgcccttg gtgccgatgc
600 agcaggggcg gcccttgatc tcgcagtcca tgtgcaggaa gcctgtgtgg
ttgctcctgg 660 cctgctctgt gcagatcggc cacttagtga tgtcatctcg tgcc
704
* * * * *