JP2002000279A - Ys 68 gene participating in primary hemopoiesis - Google Patents

Abstract

PROBLEM TO BE SOLVED: To obtain a new protein participating in primary hemopoiesis and a gene encoding the protein and provide their production and use. SOLUTION: The new gene (YS 68) participating in primary hemopoiesis is successfully isolated from a mouse yolk sac-derived cDNA. Further, the isolation of the human gene correspondent to the gene is successfully carried out. From the expression characteristic, it is suggested that these genes participate in the primary hemopoiesis. The protein and the gene encoding the protein can be utilized as a tool for developing medicines against diseases such as hemopathy and the like.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

TECHNICAL FIELD The present invention relates to a novel protein involved in early hematopoiesis and its gene. These molecules can be used, for example, in the field of drug development.

[0002]

2. Description of the Related Art Hematopoiesis includes transient fetal hematopoiesis that functions only during the fetal period and definitive hematopoiesi that contributes to life-long hematopoiesis.
s). Fetal hematopoiesis occurs in the yolk sac around embryonic day 9, whereas adult hematopoiesis occurs in the AGM region (Aorta-G) around embryonic day 10.
A study by Medinsky, et al. (Medvinsky, A., and Dzierzak, E. (1996) Cell,
86, 897-906; Cumano, A. et al., Cell, 86, 907-91
6). In addition, various studies on the origin of blood cells suggest that adult hematopoiesis is derived from hemangioblasts, which are considered to be common progenitors of hematopoietic cells and vascular endothelial cells.

[0003] Hemangioblast, which is the source of adult hematopoiesis, is predominantly present in the AGM region.
der and colleagues have shown that the yolk sac also contains hemangioblasts that can contribute to adult hematopoiesis (Yoder, MC et al., (1997) Immunity, 7,
335-344). For this reason, it has become common to think that the surrounding environment is important for hemangioblast to differentiate into hematopoietic cells.

[0004] As described above, the origin of hematopoietic cells and the place of development are gradually being clarified from phenomenological studies, but on the other hand, the molecular mechanisms of hematopoiesis development are hardly understood. The isolation of new molecules involved in early hematopoiesis will be an important step for the development of unprecedented pharmaceuticals.

[0005]

SUMMARY OF THE INVENTION An object of the present invention is to provide a novel protein involved in early hematopoiesis, its gene, and its production and use.

[0006]

[Means for Solving the Problems] Hemangioblasts are used for mouse AGM (Aorta-Gonad-Mesonephro
s) Although its presence has been reported in the area, Yorder and Nishikawa et al. report that hemangioblasts are present in the yolk sac on embryonic day 9 but are no longer present on embryonic day 13 ( Yoder, MC et al., (1997) Immunity, 7, 335-344;
Nishikawa, SI etal., (1998) Immunity, 8, 761-76
9). The present inventors, for the purpose of searching for molecules involved in the early stage of hematopoiesis, from the yolk sac-derived cDNA of mouse embryonic day 9 suggesting the presence of hemangioblasts, hemangioblasts are not present. The gene was cloned by subtracting the cDNA derived from the yolk sac of the 13th embryo, and a new gene named "YS68" was successfully isolated. In addition, primers were designed based on the nucleotide sequence of the mouse gene, and human fetal liver Marathon-Ready
By performing 5'-RACE and 3'-RACE methods using cDNA as a template, the corresponding human gene was successfully isolated.

[0007] Human and mouse full-length cDNA sequences are determined,
As a result of comparison, the N-terminal region (human 1-1137, mouse 1-1137) had a very high homology of 87%, but the central region (human 1138-1683, mouse 1138-1679) had 57% homology. And C
The terminal region (human 1684-2266, mouse 1680-2243) showed a very low homology of 45%. There are multiple nuclear localization signals in the C-terminal region with low homology, while N
In the terminal region, there were two WD repeats known to be required for the interaction between proteins.

To investigate the role of “YS68” in hematopoiesis, the expression pattern of “YS68” in mouse hematopoietic tissues was examined.
As a result of analysis by RT-PCR, "YS68" showed an expression pattern correlated with the migration of hematopoietic tissues during the fetal period. Also, "YS68"
Was expressed in CD34-positive undifferentiated blood cells. This suggests that “YS68” has an important function for early hematopoiesis.

[0009] From experiments using a yeast system, human
It was revealed that a transcription activation region exists in the region from position 1080 to position 1630 of YS68. Therefore, it was suggested that YS68 functions as a transcription factor in cells. further,
From experiments using the two-hybrid system, the region from position 35 to position 604 of human YS68 was found to be protein kinase C inhibitor 1 (PKC
I), two ESTs, and CD27 binding protein (siva1) and its mutants.

[0010] The "YS68" protein of the present invention is useful as a tool for elucidating the mechanism of early hematopoiesis, and is also expected to be applied to drug development for various diseases involving the hematopoietic system.

The present invention relates to a novel "YS6" involved in early hematopoiesis.
8 ”Regarding proteins and their genes, and their production and use, more specifically, (1) the following (a)
To (d), (a) SEQ ID NO:
DNA encoding a protein consisting of the amino acid sequence of 12 or 14, (b) SEQ ID NO: 11 or 13
A DNA containing the coding region of the nucleotide sequence described in SEQ ID NO: 12 or 14;
Or, a plurality of amino acids are substituted, deleted, inserted, and / or
Or consisting of an added amino acid sequence, SEQ ID NO: 12
Or a DNA encoding a protein functionally equivalent to the protein consisting of the amino acid sequence of 14; (d) DNA consisting of the nucleotide sequence of SEQ ID NO: 11 or 13
That hybridizes under stringent conditions with DNA
DNA encoding a protein functionally equivalent to the protein consisting of the amino acid sequence of SEQ ID NO: 12 or 14, (2) a partial peptide of the protein consisting of the amino acid sequence of SEQ ID NO: 12 or 14 (3) a protein or peptide encoded by the DNA according to (1) or (2),
(4) A vector into which the DNA according to (1) or (2) has been inserted, (5) a host cell carrying the vector according to (4), and (6) a host cell according to (5). ,
(3) The method for producing a protein or peptide according to (3), comprising a step of recovering a protein expressed from the host cell or a culture supernatant thereof, (7) DNA comprising the nucleotide sequence of SEQ ID NO: 11 or 13. Or a polynucleotide comprising at least 15 nucleotides complementary to a complementary strand thereof, (8) a method for screening a compound that binds to a protein according to (3), wherein (a) a test sample for the protein or a partial peptide thereof; (B) detecting the binding activity between the protein or its partial peptide and a test sample, and (c) selecting a compound having an activity to bind to the protein or its partial peptide. A method, (9) a compound that binds to the protein according to (3), (10) an antibody,
(9) the compound according to (11), which can be isolated by the method according to (8), which binds to the protein according to (3), (12) the protein according to (3), and PKCI or siva1 A method of screening a compound that regulates the binding to PKCI or siva1 in the presence of a test sample, and a protein or a partial peptide thereof according to (3), (b) (13) (3) a method comprising the steps of: detecting the binding; and (c) selecting a compound having an activity of promoting or inhibiting these bindings as compared to the case where the binding is detected in the absence of the test sample. A) a compound that regulates the binding of the protein described in (3) to PKCI or siva1; (14) a compound that regulates the binding of the protein described in (3) to PKCI or siva1 which can be isolated by the method described in (13) Compounds You.

[0012]

BEST MODE FOR CARRYING OUT THE INVENTION The present invention provides a novel protein involved in early hematopoiesis and a DNA encoding the protein. Mouse “YS68” isolated by the present inventors
SEQ ID NO: 11 shows the nucleotide sequence of the full-length cDNA of SEQ ID NO: and the amino acid sequence of the protein encoded by the cDNA.
FIG. In addition, the nucleotide sequence of the full-length cDNA of human “YS68” isolated by the present inventors is shown in SEQ ID NO: 13,
The amino acid sequence of the protein encoded by the cDNA is shown in SEQ ID NO: 14.

Hematopoietic stem cells that contribute to life-long hematopoiesis can be differentiated from hemangioblasts, which are common mother cells of blood cells and blood vessels. Recent gene disruption experiments have reported several transcription factors that are considered important for early hematopoiesis. SCL disrupted mice no longer show hematopoiesis as well as angiogenesis (Porcher C, et al., (1996) C
ell. 86, 47-57, Visvader JE, et al., (1998) Genes D.
ev. 12, 473-479). In addition, AML-1 and c-Myb knockout mice have no abnormality in angiogenesis, but completely lose adult hematopoiesis (Okuda T, et al., (1996) Cell. 84,
321-330, LinHH, et al., (1996) Curr Top Microbiol
Immunol. 211, 79-87). However, each of these transcription factors interacts at the stage of early hematopoiesis,
Little is known about its involvement in cell fate decisions.

The mouse "YS6" identified by the present inventors.
The `` 8 '' gene is derived from the yolk sac from mouse embryonic day 9 mouse embryos suggesting the presence of hemangioblasts,
Isolated by subtracting NA. The isolated mouse "YS68" gene was predicted to encode a protein of 1265 amino acids, and its expression was high in the yolk sac on embryonic day 9 and showed a slowly decreasing expression pattern thereafter. In the AGM region, which is considered to be a site of hematopoietic stem cell development, expression was observed from embryonic day 10, and the embryo was strongly expressed in the liver on day 13 and in the thymus and spleen on embryonic day 16 instead. Expression at these sites was significantly reduced in adult mice. YS cloned by the present inventors
68 "could be newly added as one of the molecular members involved in early hematopoiesis due to its expression pattern during development.

[0015] "YS68" is expected to be a nuclear protein because its C-terminal region has a plurality of nuclear translocation signals, but it is strongly expressed not only in the nucleus but also around the nucleus in hepatocytes. Was observed (Example 6). Since the WD repeat required for protein binding was present on the N-terminal side and co-precipitated with multiple proteins by immunoprecipitation (Example 4), it was transferred to the nucleus by interaction with other proteins. Is suggested to be regulated.

[0016] The concept of the generation of blood cells from the vascular endothelium has existed for a relatively long time, but it has only been proven very recently. Jaffredo et al. Stain whole chicken blood vessels with fluorescent LDL,
Stained vascular endothelium was found to differentiate into blood cells (Jaffredo T, et al., (1998) Development. 125,
4575-4783). Also, Hara et al. Converted cells in the AGM region to PCLP-1 (P
Hemangioblasts were found to be enriched by sorting with odocalyxin-like Protein 1). In fact, PCLP-1 expression in the AGM region was localized to the vascular endothelium, suggesting that hemangioblasts are localized to the vascular endothelium (Hara T, et al., (1999). .
Immunity. 11, 567-578). As shown in Example 5, YS68
The expression site in the AGM region was exactly the same vascular endothelium as PCLP-1. This expression pattern is also the same for AML-1 and SCL, which are known to be important for early hematopoiesis. In addition, considering that the expression of YS68 in blood cells was strong in CD34-positive cells considered to be a more immature blood cell population (Example 6), YS68 functions in the process of differentiation from hemangioblast to blood cells. It is suggested that

The "YS68" protein of the present invention and the DNA encoding the protein are useful as factors and differentiation markers for controlling the development and differentiation of hematopoietic stem cells and hematopoietic functions, as well as diseases involving the protein of the present invention. It is thought that the application to diagnosis, prevention, and treatment of is possible. In current medical care, there is no means for artificially expanding hematopoietic stem cells. Hematopoietic stem cells are artificially expanded in vitro by forcing hemangioblasts, the origin of hematopoietic cells, to express YS68 with a viral vector, or by administering a compound or cytokine that can induce YS68 expression. May be able to.
Thus, it seems that YS68 may be applied to medical treatment as a new alternative to bone marrow transplantation.

Many human blood cell tumors, such as myeloid leukemia and lymphocytic leukemia, are often caused by abnormal transcription factors, and the human "YS68" gene of the present invention is also a causative gene for those diseases. There is a possibility. Therefore, in particular, human “YS68” may be applicable to gene therapy or genetic diagnosis for these diseases. In addition, drug development targeting the human YS68 gene and protein itself, or a molecule that regulates them, or a molecule or gene that is regulated by the human YS68 protein, is useful for treating or preventing the above diseases. Conceivable.

The present invention also includes proteins that are functionally equivalent to the “YS68” protein (SEQ ID NOs: 12, 14). Such proteins include, for example, "YS68"
Variants of the protein are included. In the present invention, “functionally equivalent” indicates that the target protein has at least one of the following functions. Functions that regulate the development and / or differentiation of hematopoietic cells Other proteins (eg, protein kinase C inhibit
or 1 (PKCI); Razin E. et al., J. Biol. Chem. vol.2
74, 34272-34276 (1999), CD27 binding protein (siva1
Or its variants); Yoon Y. et al., Oncogene. Vol. 1
8, 7174-7179 (1999)). Transcription activating ability. Whether a protein has a function of controlling the development and / or differentiation of hematopoietic cells is determined by, for example, hematopoiesis as described in Example 2. Expression characteristics in tissues can be used as an index. Whether or not the test protein has the function of interacting with another protein can be determined using, for example, immunoprecipitation as described in Example 4. Further, the interaction with other proteins can be determined using a yeast two-hybrid system (see Example 9). The transcription activity of the test protein can be determined, for example, by the experimental method described in Example 8.

As a method well known to those skilled in the art for preparing a protein functionally equivalent to a certain protein, a method for introducing a mutation into a protein is known.
For example, those skilled in the art will recognize site-directed mutagenesis (Hash
imoto-Gotoh, T. et al. (1995) Gene 152, 271-275, Z
oller, MJ, and Smith, M. (1983) Methods Enzymol. 10
0, 468-500, Kramer, W. et al. (1984) Nucleic Acids
Res. 12, 9441-9456, Kramer W, and Fritz HJ (1987)
Methods.Enzymol. 154, 350-367, Kunkel, TA (1985) Pr.
oc Natl Acad Sci US A. 82, 488-492, Kunkel (1988)
Methods Enzymol. 85, 2763-2766) and the like.
By appropriately introducing mutations into the amino acids of the “S68” protein (SEQ ID NOs: 12 and 14), a protein functionally equivalent to the protein can be prepared. Also,
Amino acid mutations can also occur in nature. Thus, a protein having an amino acid sequence in which one or more amino acids are mutated in the amino acid sequence of the “YS68” protein (SEQ ID NOs: 12, 14) and functionally equivalent to the protein is also included in the protein of the present invention. included. The number of amino acids to be mutated in such a mutant is usually
100 amino acids or less, preferably 50 amino acids or less, more preferably 20 amino acids or less, further preferably 10 amino acids or less, more preferably
It is considered to be within 5 amino acids.

The amino acid residue to be mutated is desirably mutated to another amino acid whose amino acid side chain properties are conserved. For example, the properties of amino acid side chains include hydrophobic amino acids (A, I, L, M, F, P, W, Y, V),
Hydrophilic amino acids (R, D, N, C, E, Q, G, H, K, S,
T), amino acids having aliphatic side chains (G, A, V, L, I,
P), amino acids having hydroxyl group-containing side chains (S, T, Y), amino acids having sulfur atom-containing side chains (C, M), amino acids having carboxylic acid and amide-containing side chains (D, N, E, Q) ),
Examples of amino acid having a base-containing side chain (R, K, H) and amino acid having an aromatic-containing side chain (H, F, Y, W) can be given. ).

It is already known that a protein having an amino acid sequence modified by deletion, addition and / or substitution of one or more amino acid residues to a certain amino acid sequence maintains its biological activity. (Mark, DF et al., Proc. Natl. Acad. Sci. US
A (1984) 81, 5662-5666, Zoller, MJ & Smith, M.
Nucleic Acids Research (1982) 10, 6487-6500, Wan
g, A. et al., Science 224, 1431-1433, Dalbadie-Mc
Farland, G. et al., Proc. Natl. Acad. Sci. USA (19
82) 79, 6409-6413).

A protein in which a plurality of amino acid residues are added to the amino acid sequence of the “YS68” protein includes “YS6
Fusion proteins including the 8 "protein are included. The fusion protein is a fusion of the “YS68” protein and another peptide or protein, and is included in the present invention. The fusion protein is prepared by ligating the DNA encoding the “YS68” protein (SEQ ID NOs: 12 and 14) and the DNA encoding the other peptide or protein so that their frames match, and introducing this into an expression vector. And
What is necessary is just to express in a host, The technique well-known to those skilled in the art can be used. Other peptides or proteins to be fused with the protein of the present invention are not particularly limited.

Other peptides to be fused with the protein of the present invention include, for example, FLAG (Hopp, TP et a
l., BioTechnology (1988) 6, 1204-1210), 6 Hiss
6 × His, 10 × His consisting of (histidine) residues, influenza agglutinin (HA), human c-myc fragment, VSV-GP fragment, p18HIV fragment, T7-tag, HSV-tag, E-tag , SV40T antigen fragment, lck tag, α-tubulin fragment, B-tag, Prote
A known peptide such as a fragment of in C can be used. Other proteins to be fused with the protein of the present invention include, for example, GST (glutathione-S-
Transferase), HA (influenza agglutinin),
Immunoglobulin constant region, β-galactosidase, MB
P (maltose binding protein) and the like. A fusion protein can be prepared by fusing a commercially available DNA encoding the peptide or protein with a DNA encoding the protein of the present invention, and expressing the fusion DNA prepared thereby.

Further, a protein in which a plurality of amino acid residues are added to the amino acid sequence of the “YS68” protein includes:
“A” from position 98 to 692 in the nucleotide sequence of SEQ ID NO: 15
The protein encoded by the nucleotide sequence up to “g” at position 2 (the amino acid sequence represented by “M
et Ala Ala Glu Arg Arg Cys Gly Ser ").

Other methods well known to those skilled in the art for preparing proteins functionally equivalent to a protein include hybridization techniques (Sambrook, J et al.).
l., Molecular Cloning 2nd ed., 9.47-9.58, Cold Spr
ing Harbor Lab. press, 1989). That is, those skilled in the art can isolate a highly homologous DNA based on the DNA sequence encoding the “YS68” protein (SEQ ID NOs: 11 and 13) or a part thereof,
Isolation of a protein functionally equivalent to the “YS68” protein from the DNA can usually be performed. The present invention includes proteins encoded by DNA that hybridizes under stringent conditions with DNA encoding the “YS68” protein and functionally equivalent to the “YS68” protein. When isolating a cDNA highly homologous to the DNA encoding the “YS68” protein, fetal hematopoietic tissues (eg, yolk sac in early development, AGM regions, liver, thymus, spleen, etc. during the middle and late stages of development) Tissue) is considered preferable.

Hybridization conditions for isolating a DNA encoding a protein functionally equivalent to the “YS68” protein can be appropriately selected by those skilled in the art. Hybridization conditions include, for example, low stringency conditions. The low stringent conditions are, for example, conditions of 42 ° C., 2 × SSC, 0.1% SDS, preferably 50 ° C., 2 × SSC, 0.1% SDS in washing after hybridization.
More preferred hybridization conditions include:
Highly stringent conditions may be mentioned. High stringency conditions include, for example, 65 ° C., 0.1 × SSC and 0.1% SDS
Is the condition. Under these conditions, it can be expected that DNA with higher homology can be obtained more efficiently as the temperature is increased. However, a plurality of factors such as temperature and salt concentration can be considered as factors affecting the stringency of hybridization, and those skilled in the art can realize similar stringency by appropriately selecting these factors. .

Also, instead of hybridization,
DNA encoding “YS68” protein (SEQ ID NO: 1)
1, 13) gene amplification using primers synthesized based on the sequence information, for example, polymerase chain reaction (PC
It is also possible to isolate using the R) method.

The DNA isolated by these hybridization techniques and gene amplification techniques encodes “YS6
Proteins that are functionally equivalent to the "8" protein are usually
It has high homology in amino acid sequence with the “YS68” protein (SEQ ID NOs: 12, 14). The protein of the present invention also includes a protein that is functionally equivalent to the “YS68” protein and has high homology to the amino acid sequence shown in SEQ ID NO: 12 or 14. High homology usually means 30% or more identity, preferably 50% or more identity, more preferably 70% or more identity, more preferably 90% or more identity (for example, 95% or more Identity). To determine protein homology, refer to the literature (Wilbur, WJ and Lipman, DJ Proc. Natl. Aca
d. Sci. USA (1983) 80, 726-730).

The protein of the present invention may vary in amino acid sequence, molecular weight, isoelectric point, presence / absence and form of sugar chains, etc., depending on the cell, host, or purification method that produces the protein as described below. However, the obtained protein is "YS68"
As long as it has a function equivalent to a protein, it is included in the present invention. For example, when the protein of the present invention is expressed in a prokaryotic cell, for example, Escherichia coli, a methionine residue is added to the N-terminal of the amino acid sequence of the original protein. The proteins of the present invention also include such proteins.

The protein of the present invention can be prepared as a recombinant protein or a natural protein by methods known to those skilled in the art. DNA that encodes the protein of the present invention if it is a recombinant protein
(For example, a DNA having the nucleotide sequence of SEQ ID NO: 11 or 13) was incorporated into an appropriate expression vector, and the resulting transformant was introduced into an appropriate host cell to collect a transformant, thereby obtaining an extract. Thereafter, ion exchange, reverse phase, chromatography such as gel filtration, or subjected to affinity chromatography in which the antibody against the protein of the present invention is immobilized on a column, or further purified by combining a plurality of these columns, prepared It is possible to

Further, the protein of the present invention is obtained by using glutathione
When expressed in host cells (eg, animal cells, E. coli, etc.) as a fusion protein with S-transferase protein or as a recombinant protein to which multiple histidines are added, the expressed recombinant protein is a glutathione column or a nickel column. Can be used for purification.

After purifying the fusion protein, if necessary, regions of the fusion protein other than the target protein may be
It is also possible to cut and remove with thrombin or Factor Xa.

In the case of a natural protein, an antibody that binds to the protein of the present invention described below binds to a method known to those skilled in the art, for example, an extract of a tissue or a cell expressing the protein of the present invention. It can be isolated by purification by the action of an affinity column. Antibodies may be polyclonal or monoclonal.

The present invention also includes a partial peptide of the protein of the present invention. The partial peptide of the present invention has an amino acid sequence of at least 7 amino acids or more, preferably 8 amino acids or more, more preferably 9 amino acids or more. The partial peptide can be used, for example, for preparing an antibody against the protein of the present invention, screening for a compound that binds to the protein of the present invention, and screening for an accelerator or inhibitor of the protein of the present invention. In addition, it can be an antagonist or a competitive inhibitor of the protein of the present invention.
The partial peptide of the present invention includes a partial peptide containing a transcription activation region of the protein of the present invention (for example, SEQ ID NO: 1).
4 including a partial peptide including positions 1080 to 1630) and a peptide including an interaction region of the protein of the present invention with other proteins (eg, a partial peptide including positions 35 to 604 of SEQ ID NO: 14). .

The partial peptide of the present invention can be produced by genetic engineering techniques, known peptide synthesis methods, or by cleaving the protein of the present invention with an appropriate peptidase. The peptide may be synthesized by, for example, either a solid phase synthesis method or a liquid phase synthesis method.

The DNA encoding the protein of the present invention is
As described above, the in vivo or in v
In addition to being used for production in itro, for example, application to gene therapy of diseases caused by abnormalities in the gene encoding the protein of the present invention or diseases treatable by the protein of the present invention can be considered. The DNA of the present invention may be in any form as long as it can encode the protein of the present invention. That is, cDNA synthesized from mRNA or genomic
It does not matter whether it is DNA or chemically synthesized DNA. In addition, DNAs having any base sequence based on the degeneracy of the genetic code are included as long as they can encode the protein of the present invention.

The DNA of the present invention can be prepared by a method known to those skilled in the art. For example, it is prepared by preparing a cDNA library from cells expressing the protein of the present invention and performing hybridization using a part of the sequence of the DNA of the present invention (for example, SEQ ID NOs: 11 and 13) as a probe. it can. cDNA libraries are described, for example, in the literature (Sambrook, J. et al., Molecular Cloning, Cold S.
Spring Harbor Laboratory Press (1989)), or a commercially available DNA library may be used. Further, after preparing RNA from cells expressing the protein of the present invention and synthesizing cDNA by reverse transcriptase, the sequence of the DNA of the present invention (for example, SEQ ID NO: 11,
It can also be prepared by synthesizing an oligo DNA based on 13), performing a PCR reaction using this as a primer, and amplifying a cDNA encoding the protein of the present invention.

By determining the nucleotide sequence of the obtained cDNA, the translation region encoded by the cDNA can be determined, and the amino acid sequence of the protein of the present invention can be obtained.
Genomic DNA can be isolated by screening a genomic DNA library using the obtained cDNA as a probe.

More specifically, the following may be performed. First, cells, tissues, and organs that express the protein of the present invention
MRNA is isolated from (eg, yolk sac in early development, AGM regions, fetal hematopoietic tissues such as liver, thymus, spleen, etc. from mid to late development). mRNA can be isolated by known methods, for example, guanidine ultracentrifugation (Chirgwin, JM et al.).
al., Biochemistry (1979) 18, 5294-5299), AGPC method
(Chomczynski, P. and Sacchi, N., Anal.Biochem. (19
87) Prepare total RNA according to 162, 156-159)
mRNA from total RNA using fication Kit (Pharmacia)
Is purified. Also, QuickPrep mRNA Purification Kit
(Pharmacia) to directly prepare mRNA.

From the obtained mRNA, cDNA was prepared using reverse transcriptase.
Are synthesized. For cDNA synthesis, AMV Reverse Transcriptas
e It can also be performed using a First-strand cDNA Synthesis Kit (Seikagaku Corporation) or the like. In addition, using the primers and the like described in this specification, 5'-Ampli FINDER RACE Kit
(Clontech) and polymerase chain reaction (polymerase
5'-RACE method (Frohman,
MA et al., Proc. Natl. Acad. Sci. USA (1988)
85, 8998-9002; Belyavsky, A. et al., Nucleic Aci
ds Res. (1989) 17, 2919-2932), cDNA can be synthesized and amplified.

A target DNA fragment is prepared from the obtained PCR product and ligated to a vector DNA. Further, a recombinant vector is prepared from this, introduced into E. coli, etc., and colonies are selected to prepare a desired recombinant vector. Aim
The DNA base sequence can be confirmed by a known method, for example, a dideoxynucleotide chain termination method.

In the DNA of the present invention, a nucleotide sequence having higher expression efficiency can be designed in consideration of the codon usage of the host used for expression (Grantham, R., et al.
etal., Nucelic Acids Research (1981) 9, r43-74
). The DNA of the present invention can be modified by a commercially available kit or a known method. Modifications include, for example, digestion with restriction enzymes, insertion of synthetic oligonucleotides or appropriate DNA fragments, addition of linkers, initiation codon (ATG) and / or termination codon (TAA, TGA, or TA
G) insertion and the like.

The DNA of the present invention is a DNA which hybridizes under stringent conditions with a DNA consisting of the nucleotide sequence shown in SEQ ID NO: 11 or 13, and which is functionally equivalent to the protein of the present invention. DNA that encodes Stringent conditions for hybridization can be appropriately selected by those skilled in the art, and specifically, the conditions described above can be used. Under these conditions, DNA with higher homology can be obtained as the temperature is increased. The DNA to be hybridized is preferably a DNA of natural origin, for example,
cDNA or chromosomal DNA.

The present invention also provides a vector into which the DNA of the present invention has been inserted. The vector of the present invention is useful for retaining the DNA of the present invention in a host cell or expressing the protein of the present invention.

For example, when E. coli is used as a host, the vector may be E. coli (for example, JM109, D
H5α, HB101, XL1Blue) for large-scale amplification and preparation, have an “ori” to be amplified in E. coli, and further select genes for transformed E. coli (for example, any drug (ampicillin, tetracycline, There is no particular limitation as long as it has a drug resistance gene which can be distinguished by kanamycin or chloramphenicol). Examples of vectors include M13-based vectors, pUC-based vectors, pBR322, pBluescript, pCR-Script, and the like. In addition, in the case of subcloning and excision of cDNA, in addition to the above vectors, for example, pGEM-T, pD
IRECT, pT7 and the like. When a vector is used for the purpose of producing the protein of the present invention, an expression vector is particularly useful. As an expression vector, for example, when the purpose is expression in E. coli, in addition to having the above-mentioned characteristics such that the vector is amplified in E. coli, the host was JM109, DH5α, HB101, E. coli such as XL1-Blue. In some cases, a promoter that can be efficiently expressed in E. coli, such as the lacZ promoter (Ward
Et al., Nature (1989) 341, 544-546; FASEB J. (1992) 6,
2422-2427), araB promoter (Better et al., Science
(1988) 240, 1041-1043), or have a T7 promoter. Such vectors include pGEX-5X-1 (Pharmacia), “QIAexpress system” (Qiagen), p
EGFP, or pET (in this case, the host is preferably BL21 expressing T7 RNA polymerase).

Further, the vector may contain a signal sequence for polypeptide secretion. As a signal sequence for protein secretion, a pelB signal sequence (Lei, SP
et al J. Bacteriol. (1987) 169, 4379) may be used. Introduction of a vector into a host cell can be performed using, for example, a calcium chloride method or an electroporation method.

In addition to Escherichia coli, for example, as a vector for producing the protein of the present invention, a mammalian expression vector (for example, pcDNA3 (manufactured by Invitrogen) or pEGF-BOS (Nucleic Acids. Res. 1990, 18 (17),
p5322), pEF, pCDM8), insect cell-derived expression vectors (eg, “Bac-to-BAC baculovairus expression system”
m "(manufactured by Gibco BRL), pBacPAK8), plant-derived expression vectors (eg, pMH1, pMH2), animal virus-derived expression vectors (eg, pHSV, pMV, pAdexLcw), retrovirus-derived expression vectors (eg, pZIPneo) ), Yeast-derived expression vectors (eg, “Pichia Expression Ki
t "(manufactured by Invitrogen), pNV11, SP-Q01), and an expression vector derived from Bacillus subtilis (for example, pPL608, pKTH50).

For the purpose of expression in animal cells such as CHO cells, COS cells, and NIH3T3 cells, promoters necessary for expression in cells, for example, SV40 promoter (Mulligan et al., Nature (1979) 277, 108), MMLV-LTR promoter, EF1α promoter (Mizushima et al., Nuclei
c Acids Res. (1990) 18, 5322), it is essential to have a CMV promoter, etc., so that it can be distinguished by a gene (for example, a drug (neomycin, G418, etc.)) for selecting cell transformation. Is more preferable. Vectors having such properties include, for example, pMAM, pDR2, pBK-RSV, p
BK-CMV, pOPRSV, pOP13 and the like.

When the gene is to be stably expressed and the gene copy number is to be amplified in a cell, the CHO cell lacking the nucleic acid synthesis pathway has a DHFR gene that complements it. A method of introducing a vector (for example, pCHOI) and amplifying it with methotrexate (MTX) can be mentioned. In the case of transient expression of a gene, a gene expressing the SV40 T antigen is transferred onto the chromosome. And a method of transforming a vector having an SV40 origin of replication (such as pcD) using the COS cell contained in the above. As the replication origin, those derived from polyoma virus, adenovirus, bovine papilloma virus (BPV) and the like can also be used. Furthermore, in order to amplify the gene copy number in the host cell system, the expression vector is used as a selection marker as an aminoglycoside transferase (APH) gene, thymidine kinase (TK) gene, Escherichia coli xanthine guanine phosphoribosyl transferase (Ecogp
t) gene, dihydrofolate reductase (dhfr) gene and the like.

On the other hand, as a method for expressing the DNA of the present invention in an animal body, the DNA of the present invention is incorporated into an appropriate vector, for example, a retrovirus method, a liposome method, or the like.
Examples thereof include a method of introducing into a living body by a cationic liposome method, an adenovirus method, and the like. This makes it possible to perform gene therapy for a disease caused by a mutation in the “YS68” gene of the present invention. As a vector to be used, for example, an adenovirus vector (for example, pAdexlcw) or a retrovirus vector (for example, pZIPn
eo) and the like, but are not limited thereto. General gene manipulation such as insertion of the DNA of the present invention into a vector can be performed according to a conventional method (Molecular Cl
oning, 5.61-5.63). Administration into a living body may be an ex vivo method or an in vivo method.

[0052] The present invention also relates to a host cell into which the vector of the present invention has been introduced. The host cell into which the vector of the present invention is introduced is not particularly limited, and for example, Escherichia coli and various animal cells can be used. The host cell of the present invention can be used, for example, as a production system for producing or expressing the protein of the present invention. Production systems for protein production include in vitro and in vivo production systems. Examples of the in vitro production system include a production system using eukaryotic cells and a production system using prokaryotic cells.

When eukaryotic cells are used, for example, animal cells, plant cells, and fungal cells can be used as hosts.
As animal cells, mammalian cells, for example, CHO (J. Exp.
Med. (1995) 108, 945), COS, 3T3, myeloma, BHK
(Baby hamster kidney), HeLa, Vero, amphibian cells,
For example, Xenopus oocytes (Valle, et al., Na
(1981) 291, 358-340), or insect cells such as Sf9, Sf21, and Tn5. As the CHO cells, particularly, dhfr-CH, which is a CHO cell deficient in the DHFR gene,
O (Proc. Natl. Acad. Sci. USA (1980) 77, 4216-422
0) and CHO K-1 (Proc. Natl. Acad. Sci. USA (1968) 6
0, 1275) can be suitably used. In the case of aiming for large-scale expression in animal cells, CHO cells are particularly preferable. Introduction of a vector into a host cell can be performed by, for example, a calcium phosphate method, a DEAE dextran method, a method using cationic ribosome DOTAP (manufactured by Boehringer Mannheim), an electroporation method, or a lipofection method.

As plant cells, for example, Nicotiana
Cells derived from tabacum (Nicotiana tabacum) are known as a protein production system, which may be callus cultured. As fungal cells, yeasts, for example, genus Saccharomyces, for example, Saccharomyces cerevisiae, and filamentous fungi, for example, genus Aspergillus, for example, Aspergillus niger are known. .

When using prokaryotic cells, there is a production system using bacterial cells. Bacterial cells include E. coli (E. col.
i), for example, JM109, DH5α, HB101 and the like, and Bacillus subtilis are also known.

The protein can be obtained by transforming these cells with the desired DNA and culturing the transformed cells in vitro. Culture can be performed according to a known method. For example, as a culture solution of animal cells, for example, DMEM, MEM, RPMI1640, IMDM can be used. At that time, a serum replacement solution such as fetal calf serum (FCS) may be used in combination, or serum-free culture may be performed. The pH during the culturing is preferably about 6-8. Culture is usually performed at about 30 to 40 ° C. for about 15 to 200 hours, and the medium is replaced, aerated, and agitated as necessary.

On the other hand, examples of a system for producing a protein in vivo include a production system using animals and a production system using plants. The target DNA is introduced into these animals or plants, and proteins are produced and recovered in the animals or plants. The “host” in the present invention includes these animals and plants.

When animals are used, there are production systems using mammals and insects. Goats, pigs, sheep, mice, and cows can be used as mammals (Vick
i Glaser, SPECTRUM Biotechnology Applications, 199
3). When a mammal is used, a transgenic animal can be used.

For example, the DNA of interest is prepared as a fusion gene with a gene encoding a protein that is specifically produced in milk, such as goat β-casein. Then
A DNA fragment containing the fusion gene is injected into a goat embryo, and the embryo is transplanted into a female goat. A target protein can be obtained from milk produced by a transgenic goat born from a goat that has received an embryo or a progeny thereof. Hormones may optionally be used in transgenic goats to increase the amount of milk containing proteins produced by the transgenic goats (Ebert, KM et al., Bio /
Technology (1994) 12, 699-702).

As insects, for example, silkworms can be used. When a silkworm is used, the target protein can be obtained from the body fluid of the silkworm by infecting the silkworm with a baculovirus into which DNA encoding the target protein has been inserted (Susumu, M. et al.,
Nature (1985) 315, 592-594).

Further, when using a plant, for example, tobacco can be used. When tobacco is used, DNA encoding the protein of interest is inserted into a plant expression vector, for example, pMON530, and this vector is inserted into Agrobacterium tumefaciens.
ciens). This bacterium is transferred to tobacco, for example, Nicotian Tabacum.
a tabacum) to obtain the desired polypeptide from the leaves of this tobacco (Julian K.-C. Maet al.,
Eur. J. Immunol. (1994) 24, 131-138).

The thus-obtained protein of the present invention is isolated from the inside or outside of a host cell (such as a medium),
It can be purified as a substantially pure and homogeneous protein. The separation and purification of the protein may be carried out by the separation and purification methods used in ordinary protein purification, and are not limited at all. For example, chromatographic columns, filters, ultrafiltration, salting out, solvent precipitation, solvent extraction, distillation, immunoprecipitation, SDS-polyacrylamide gel electrophoresis, isoelectric focusing, dialysis, recrystallization, etc. When combined, proteins can be separated and purified.

Examples of the chromatography include affinity chromatography, ion exchange chromatography, hydrophobic chromatography, gel filtration, reverse phase chromatography, adsorption chromatography, and the like (Strategies for Protein Purification and Chlorography).
aracterization: A Laboratory Course Manual. Ed Dan
iel R. Marshak et al., Cold Spring Harbor Laborato
ry Press, 1996). These chromatographys can be performed using liquid phase chromatography, for example, liquid phase chromatography such as HPLC and FPLC. The present invention
Highly purified proteins using these purification methods are also included.

The protein can be arbitrarily modified or partially removed by applying an appropriate protein modifying enzyme before or after the protein is purified. As the protein modifying enzyme, for example, trypsin, chymotrypsin, lysyl endopeptidase, protein kinase, glucosidase and the like are used.

The present invention also provides an antibody that binds to the protein of the present invention. The form of the antibody of the present invention is not particularly limited, and includes a monoclonal antibody in addition to a polyclonal antibody. Also included are antisera obtained by immunizing an immunized animal such as a rabbit with the protein of the present invention, polyclonal antibodies and monoclonal antibodies of all classes, as well as human antibodies and humanized antibodies obtained by genetic recombination.

The protein of the present invention used as a sensitizing antigen for obtaining an antibody is not limited to the animal species from which it is derived, but is preferably a protein derived from a mammal, for example, a human, a mouse or a rat, and particularly preferably a protein derived from a human. preferable. A human-derived protein can be obtained using the gene sequence or amino acid sequence disclosed herein.

In the present invention, the protein used as a sensitizing antigen may be a complete protein,
Further, it may be a partial peptide of a protein. Examples of the partial peptide of the protein include an amino group (N) terminal fragment and a carboxy (C) terminal fragment of the protein. As used herein, “antibody” refers to an antibody that reacts with the full length or fragment of a protein.

The gene encoding the protein of the present invention or a fragment thereof is inserted into a known expression vector system, and the vector is used to transform a host cell described in the present specification. Fragments may be obtained by a known method, and these may be used as a sensitizing antigen. Alternatively, a cell expressing the protein, a lysate thereof, or a chemically synthesized protein of the present invention may be used as the sensitizing antigen. It is preferable that the short peptide is appropriately bound to a carrier protein such as keyhole limpet hemocyanin, bovine serum albumin, and ovalbumin to form an antigen.

As a mammal immunized with a sensitizing antigen,
Although not particularly limited, it is preferable to select in consideration of compatibility with a parent cell used for cell fusion. Generally, rodents, lagomorphs, and primates are used. .

As rodent animals, for example, mice, rats, hamsters and the like are used. As an animal of the order Lagomorpha, for example, a rabbit is used. As a primate animal, for example, a monkey is used. As the monkeys, monkeys of the lower nose (old world monkeys), for example, cynomolgus monkeys, rhesus monkeys, baboons, chimpanzees, and the like are used.

Immunization of an animal with a sensitizing antigen is performed according to a known method. As a general method, a sensitizing antigen is injected intraperitoneally or subcutaneously into a mammal. Specifically, the sensitizing antigen is diluted to an appropriate amount with PBS (Phosphate-Buffered Saline) or physiological saline or the like, and the suspension is mixed with an appropriate amount of a normal adjuvant, such as Freund's complete adjuvant, if desired. After emulsification, it is administered to mammals. Further, thereafter, it is preferable that the sensitizing antigen mixed with an appropriate amount of Freund's incomplete adjuvant is administered several times every 4 to 21 days. In addition, a suitable carrier can be used at the time of immunization of the sensitizing antigen. Immunization is performed in this manner, and it is confirmed by a conventional method that the level of a desired antibody in serum is increased.

Here, in order to obtain a polyclonal antibody against the protein of the present invention, the blood of a mammal sensitized with the antigen is taken out after confirming that the desired antibody level in the serum has increased. The serum is separated from the blood by a known method. As the polyclonal antibody, serum containing the polyclonal antibody may be used, or if necessary, a fraction containing the polyclonal antibody may be further isolated from the serum and used. For example, by using an affinity column to which the protein of the present invention is coupled, a fraction that recognizes only the protein of the present invention is obtained, and this fraction is further purified by using a protein A or protein G column. And immunoglobulin G or M can be prepared.

To obtain a monoclonal antibody, after confirming that the desired antibody level is increased in the serum of a mammal sensitized with the above antigen, the immune cells may be removed from the mammal and subjected to cell fusion. . At this time, spleen cells are particularly preferable as the immune cells used for cell fusion. As the other parent cell fused with the immune cell,
Preferably, a mammalian myeloma cell, more preferably, a myeloma cell that has acquired characteristics for selecting fused cells by a drug is used.

Cell fusion of the above-mentioned immune cells and myeloma cells is basically performed by a known method, for example, the method of Milstein et al. (Galfre, G. and Milstein, C., Methods Enzymol.
(1981) 73, 3-46).

The hybridoma obtained by cell fusion can be used in a conventional selective culture medium, for example, a HAT culture medium (a culture medium containing hypoxanthine, aminopterin and thymidine).
Selected by culturing. Culturing in the HAT culture solution is performed for a period of time sufficient to kill cells (non-fused cells) other than the target hybridoma, usually for several days to several weeks. Next, a usual limiting dilution method is performed to screen and clone a hybridoma producing the desired antibody.

In addition to obtaining the above hybridoma by immunizing an animal other than human with an antigen, human lymphocytes such as EB
Human lymphocytes infected with the virus are sensitized in vitro with a protein, a protein-expressing cell or a lysate thereof, and the sensitized lymphocytes are fused with a myeloma cell having a permanent division ability derived from a human, such as U266, and bound to the protein. A hybridoma producing a desired human antibody having activity can also be obtained (Japanese Patent Application Laid-Open No. 63-17688).

Next, the obtained hybridoma was transplanted into the peritoneal cavity of a mouse, ascites was collected from the mouse, and the obtained monoclonal antibody was subjected to, for example, ammonium sulfate precipitation, protein
It can be prepared by purification using A, protein G column, DEAE ion exchange chromatography, an affinity column to which the protein of the present invention is coupled, and the like. The antibody of the present invention is used for purification and detection of the protein of the present invention, and is also a candidate for an agonist or antagonist of the protein of the present invention. It is also conceivable to apply this antibody to antibody therapy for diseases involving the protein of the present invention. When the obtained antibody is used for the purpose of administration to the human body (antibody therapy), a human antibody or a humanized antibody is preferable in order to reduce immunogenicity.

For example, a transgenic animal having a repertoire of human antibody genes is immunized with a protein serving as an antigen, a protein-expressing cell or a lysate thereof to obtain antibody-producing cells, and a hybridoma obtained by fusing the cells with myeloma cells is used. To obtain a human antibody against the protein (International Publication Nos. WO92-03918, WO93
-2227, WO94-02602, WO94-25585, WO96-33735 and WO9
6-34096).

In addition to producing antibodies using hybridomas, cells in which immune cells such as sensitized lymphocytes that produce antibodies are immortalized with oncogenes may be used.

The thus obtained monoclonal antibody can also be obtained as a recombinant antibody produced using a gene recombination technique (for example, Borrebaeck, C. et al.
AKand Larrick, JW, THERAPEUTIC MONOCLONAL AN
TIBODIES, Published in the United Kingdom by MACMIL
LAN PUBLISHERS LTD, 1990). Recombinant antibodies are produced by cloning DNA encoding the same from immunocytes such as hybridomas or sensitized lymphocytes producing the antibodies, incorporating the DNA into an appropriate vector, and introducing this into a host. The present invention includes this recombinant antibody.

Furthermore, the antibody of the present invention may be an antibody fragment or a modified antibody thereof as long as it binds to the protein of the present invention. For example, antibody fragments include Fab, F (ab ') 2, Fv
Alternatively, a single chain Fv (scFv) in which the Fvs of the H chain and the L chain are linked by an appropriate linker (Huston, JS et al., Proc.
Acad. Sci. USA (1988) 85, 5879-5883). Specifically, the antibody is converted to an enzyme, for example, papain,
An antibody fragment is generated by treatment with pepsin, or a gene encoding these antibody fragments is constructed, introduced into an expression vector, and then expressed in a suitable host cell (for example, Co, MS et al., J. Immunol. (1994) 152, 29
68-2976; Better, M. and Horwitz, AH, Methods En
zymol. (1989) 178, 476-496; Pluckthun, A. and Ske
rra, A., Methods Enzymol. (1989) 178, 497-515; La
moyi, E., Methods Enzymol. (1986) 121, 652-663; R
ousseaux, J. et al., Methods Enzymol. (1986) 121,
663-669; Bird, RE and Walker, BW, Trends Bi
otechnol. (1991) 9, 132-137).

As the modified antibody, an antibody conjugated with various molecules such as polyethylene glycol (PEG) can be used. The “antibody” of the present invention also includes these modified antibodies. Such a modified antibody can be obtained by subjecting the obtained antibody to chemical modification. These methods are already established in this field.

Further, the antibody of the present invention can be prepared by using a chimeric antibody comprising a variable region derived from a non-human antibody and a constant region derived from a human antibody or a CDR (complementarity determining region) derived from a non-human antibody, using known techniques. It can be obtained as a humanized antibody composed of human antibody-derived FR (framework region) and constant region.

The antibodies obtained as described above can be purified to homogeneity. The separation and purification of the antibody used in the present invention may be performed by the separation and purification methods used for ordinary proteins. For example, if appropriate selection and combination of chromatography columns such as affinity chromatography, filters, ultrafiltration, salting out, dialysis, SDS polyacrylamide gel electrophoresis, isoelectric focusing, etc., the antibody can be separated and purified. Is possible (Antibodie
s: A Laboratory Manual. Ed Harlow and DavidLane,
Cold Spring Harbor Laboratory, 1988). The concentration of the antibody obtained above was measured by measuring absorbance or enzyme-linked immunosorbent assay (Enzyme-
linked immunosorbent assay (ELISA) or the like.

The columns used for affinity chromatography include a protein A column and a protein G column.
Columns. For example, as a column using a protein A column, Hyper D, POROS, Sepharose FF
(Pharmacia) and the like.

Examples of chromatography other than affinity chromatography include ion exchange chromatography, hydrophobic chromatography, gel filtration, reverse phase chromatography, and adsorption chromatography (Strategies for Protein Purification).
and Characterization: A Laboratory Course Manua
l. Ed Daniel R. Marshak et al., Cold Spring Harbor
Laboratory Press, 1996). These chromatographys can be performed using liquid phase chromatography such as HPLC and FPLC.

[0087] Methods for measuring the antigen-binding activity of the antibody of the present invention include, for example, measurement of absorbance and enzyme-linked immunosorbent assay (ELIS).
A), EIA (enzyme-linked immunosorbent assay), RIA (radioimmunoassay) or fluorescent antibody method can be used. When ELISA is used, the protein of the present invention is added to a plate on which the antibody of the present invention is immobilized, and then a sample containing the target antibody, for example, a culture supernatant of antibody-producing cells or a purified antibody is added. An enzyme, for example, a secondary antibody recognizing an antibody labeled with alkaline phosphatase or the like is added, the plate is incubated, and after washing, an antigen substrate is measured by adding an enzyme substrate such as p-nitrophenyl phosphate to measure the antigen. Binding activity can be evaluated. As the protein, a fragment of the protein, for example, a fragment consisting of its C-terminus may be used. To evaluate the activity of the antibody of the present invention, BIAcore (Pha
rmacia) can be used.

By using these techniques, the antibody of the present invention is brought into contact with a sample which is expected to contain the protein of the present invention contained in the sample, and an immune complex of the antibody and the protein is detected. Or consisting of measuring,
The method for detecting or measuring the protein of the present invention can be carried out. The method for detecting or measuring the protein of the present invention comprises:
Since the protein can be specifically detected or measured, it is useful for various experiments using the protein.

The present invention also provides a polynucleotide comprising at least 15 nucleotides complementary to a DNA encoding the “YS68” protein (SEQ ID NOs: 11, 13) or a complementary strand thereof.

Here, the “complementary strand” refers to one strand of a double-stranded nucleic acid consisting of A: T, G: C base pairs with respect to the other strand. Further, `` complementary '' is not limited to a completely complementary sequence in at least 15 contiguous nucleotide regions, at least 70%, preferably at least 80%, more preferably 90%, more preferably 95% It is only necessary to have the above homology on the base sequence. The algorithm described in the present specification may be used as an algorithm for determining homology.

Such nucleic acids include probes and primers used for detection and amplification of the DNA encoding the protein of the present invention, nucleotides or nucleotide derivatives (eg, antisense oligonucleotides) for suppressing the expression of the protein of the present invention. And ribozymes or DNAs encoding them). Such a nucleic acid can also be used for producing a DNA chip.

When used as a primer, the region on the 3 ′ side is complementary, and a restriction enzyme recognition sequence, a tag or the like can be added to the 5 ′ side.

The antisense oligonucleotide includes, for example, an antisense oligonucleotide that hybridizes at any position in the nucleotide sequence of SEQ ID NO: 11 or 13. This antisense oligonucleotide is preferably an antisense oligonucleotide for at least 15 or more consecutive nucleotides in the nucleotide sequence of SEQ ID NO: 11 or 13. More preferably, it is an antisense oligonucleotide in which at least 15 or more consecutive nucleotides contain a translation initiation codon.

As the antisense oligonucleotide, derivatives and modifications thereof can be used.
Examples of the modified form include a modified lower alkyl phosphonate such as a methylphosphonate type or an ethylphosphonate type, a phosphorothioate modified form or a phosphoroamidate modified form.

The antisense oligonucleotide is a DNA
Or not only those whose nucleotides corresponding to the nucleotides constituting the predetermined region of the mRNA are all complementary sequences,
DNA or mRNA and oligonucleotide are SEQ ID NO: 1
As long as it can specifically hybridize to the nucleotide sequence shown in 1 or 13, a mismatch of one or more nucleotides may exist.

The antisense oligonucleotide derivative of the present invention acts on a cell producing the protein of the present invention,
By binding to DNA or mRNA encoding the protein, inhibiting its transcription or translation, or promoting mRNA degradation, and suppressing the expression of the protein of the present invention, consequently the present invention Has the effect of suppressing the action of the protein.

The antisense oligonucleotide derivative of the present invention can be mixed with a suitable base material which is inactive against them to prepare an external preparation such as a liniment or a poultice. If necessary, excipients, isotonic agents, solubilizing agents, stabilizing agents, preservatives, soothing agents, etc. are added to tablets, splinters, granules, capsules, liposome capsules, injections. , A liquid preparation, a nasal drop and the like, and a lyophilized preparation. These can be prepared according to a conventional method.

The antisense oligonucleotide derivative of the present invention is applied directly to an affected area of a patient, or is applied to a patient so as to be able to reach the affected area as a result of intravenous administration or the like. Further, an antisense encapsulating material that enhances durability and membrane permeability can be used. For example, liposomes, poly-L-lysine, lipid, cholesterol, lipofectin or derivatives thereof can be mentioned.

The dosage of the antisense oligonucleotide derivative of the present invention can be adjusted appropriately according to the patient's condition, and a preferred amount can be used. For example, 0.1 to 100 mg /
kg, preferably in the range of 0.1 to 50 mg / kg.

The antisense oligonucleotides of the invention inhibit the expression of the protein of the invention and are therefore useful in inhibiting the biological activity of the protein of the invention. Further, the expression inhibitor containing the antisense oligonucleotide of the present invention is useful in that it can suppress the biological activity of the protein of the present invention.

The protein of the present invention is useful for screening for a compound that binds to the protein. That is, the protein of the present invention is brought into contact with a test sample expected to contain a compound that binds to the protein, the binding activity between the protein and the test sample is detected, and the protein binds to the protein of the present invention. It is used in a method for screening a compound that binds to the protein of the present invention, which comprises selecting a compound having activity.

The protein of the present invention used for screening may be a recombinant protein or a protein of natural origin. It may be a partial peptide. It may also be in a form expressed on the cell surface or as a membrane fraction. The test sample is not particularly limited and includes, for example, a cell extract, a cell culture supernatant, a fermentation microorganism product, a marine organism extract, a plant extract, a purified or crude protein, a peptide, a non-peptide compound, Molecular compounds and natural compounds. The protein of the present invention to be brought into contact with a test sample may be, for example, a purified protein, a soluble protein, a form bound to a carrier, a fusion protein with another protein, or a form expressed on a cell membrane. The sample can be brought into contact with a test sample as a membrane fraction.

As a method for screening a protein that binds to the protein using the protein of the present invention, for example, many methods known to those skilled in the art can be used. Such screening can be performed, for example, by immunoprecipitation. Specifically, it can be performed as follows. The gene encoding the protein of the present invention is inserted into a vector for exogenous gene expression such as pSV2neo, pcDNA I, pCD8 or the like, whereby the gene is expressed in animal cells or the like. SV40 early promoter (Rigby In Williamson (e
d.), Genetic Engineering, Vol. 3. Academic Press, L
ondon, p.83-141 (1982)), EF-1 α promoter (Kim et al. Ge
ne 91 , p.217-223 (1990)), CAG promoter (Niwa et a
l. Gene 108 , p.193-200 (1991)), RSV LTR promoter
(Cullen Methods in Enzymology 152 , p.684-704 (198
7), SR α promoter (Takebe et al. Mol. Cell. Biol.
8 , p.466 (1988)), CMV immediate early promoter (S
eed and Aruffo Proc. Natl. Acad. Sci. USA 84 , p. 33
65-3369 (1987)), SV40 late promoter (Gheysen andFi
ers J. Mol. Appl. Genet. 1 , p.385-394 (1982)), Ad
enovirus late promoter (Kaufman et al. Mol. Cell.
Biol. 9 , p. 946 (1989)), any commonly used promoter such as the HSV TK promoter may be used.

In order to express a foreign gene by introducing the gene into an animal cell, an electroporation method is used.
(Chu, G. et al. Nucl. Acid Res. 15 , 1311-1326 (19
87)), calcium phosphate method (Chen, C and Okayama, H.
Mol. Cell. Biol. 7 , 2745-2752 (1987)), DEAE dextran method (Lopata, MA et al. Nucl. Acids Res. 1
2 , 5707-5717 (1984); Sussman, DJ and Milman, G.
Mol. Cell. Biol. 4 , 1642-1643 (1985)), Lipofectin method (Derijard, B. Cell 7 , 1025-1037 (1994); Lam
b, BT et al. Nature Genetics 5 , 22-30 (1993); R
abindran, SKet al. Science 259 , 230-234 (1993))
Etc., but any method may be used.

By introducing a recognition site (epitope) of the monoclonal antibody of which specificity is known to the N-terminus or C-terminus of the protein of the present invention, the protein of the present invention can be used as a fusion protein having the recognition site of the monoclonal antibody. Can be expressed. A commercially available epitope-antibody system can be used (Experimental Medicine 13 , 85-90 (1995)). Vectors capable of expressing a fusion protein with β-galactosidase, maltose binding protein, glutathione S-transferase, green fluorescent protein (GFP), and the like via a multicloning site are commercially available.

In order to minimize the properties of the protein of the present invention by changing it into a fusion protein, a method for preparing a fusion protein by introducing only a small epitope portion consisting of several to several tens of amino acids is also available. It has been reported. For example, polyhistidine (His-ta
g), influenza agglutinin HA, human c-myc, FLAG, Ves
icular stomatitis virus glycoprotein (VSV-GP),
Bind epitopes such as T7 gene10 protein (T7-tag), human herpes simplex virus glycoprotein (HSV-tag), E-tag (epitope on monoclonal phage) and monoclonal antibodies that recognize it to the protein of the present invention. It can be used as an epitope-antibody system for protein screening (Experimental Medicine 13 , 85-90 (19
95)).

In immunoprecipitation, these antibodies are added to a cell lysate prepared using an appropriate surfactant to form an immune complex. This immune complex comprises the protein of the present invention, a protein capable of binding thereto, and an antibody. In addition to using antibodies against the above epitopes, immunoprecipitation can be performed using antibodies against the protein of the present invention. Antibodies against the protein of the present invention include, for example, a gene encoding the protein of the present invention introduced into an appropriate Escherichia coli expression vector, expressed in Escherichia coli, and the expressed protein is purified. It can be prepared by immunizing goats, chickens and the like. Alternatively, it can be prepared by immunizing the above animal with the synthesized partial peptide of the protein of the present invention.

[0108] The immune complex is, for example, the antibody is a mouse IgG.
For antibodies, use Protein A Sepharose or Protein G Se
It can be sedimented using pharose. Further, when the protein of the present invention is prepared as a fusion protein with an epitope such as GST, glutathione
-Using a substance that specifically binds to these epitopes such as Sepharose 4B, an immune complex can be formed in the same manner as when an antibody against the protein of the present invention is used.

For general methods of immunoprecipitation, see, for example, the literature (Harlow, E. and Lane, D .: Antibodies, pp. 5).
11-552, Cold Spring Harbor Laboratory publication
s, New York (1988)) or in accordance therewith.

For analysis of immunoprecipitated proteins, use SDS
-PAGE is common, and by using a gel of an appropriate concentration, the bound protein can be analyzed by the molecular weight of the protein. In addition, at this time, it is generally difficult to detect the protein bound to the protein of the present invention by ordinary staining methods such as Coomassie staining and silver staining, so that ³⁵ S-methionine which is a radioisotope is used. and ³⁵ S- cysteine cells are cultured inclusive culture medium, labeled protein of the intracellular, it is possible to improve the detection sensitivity by detecting this. Once the molecular weight of the protein is known, the protein of interest can be purified directly from the SDS-polyacrylamide gel and its sequence determined.

The present inventors have detected a plurality of proteins that bind to the protein of the present invention by immunoprecipitation in Examples (Example 4).

As a method for isolating a protein that binds to the protein using the protein of the present invention, for example, a West Western blotting method (Skolnik,
EY et al., Cell (1991) 65, 83-90) can be used. That is, cells, tissues, and organs that are expected to express a binding protein that binds to the protein of the present invention (e.g., early developmental yolk sac, AGM region, liver, thymus, spleen, etc. during the middle to late stages of development) CDNA library using phage vector (λgt11, ZAP, etc.) from fetal hematopoietic tissue of, etc.), express this on LB-agarose, fix the protein expressed on filter, purify and label The protein of the present invention thus prepared is allowed to react with the above-mentioned filter, and plaque expressing the protein bound to the protein of the present invention may be detected by labeling. Examples of the method for labeling the protein of the present invention include a method using the binding property of biotin and avidin, and an antibody that specifically binds to the protein of the present invention or a peptide or polypeptide (eg, GST or the like) fused to the protein of the present invention. , A method using a radioisotope, a method using fluorescence, and the like.

As another embodiment of the screening method of the present invention, a two-hybrid system using cells (Fields, S., and Sternglanz, R., Trends. Genet. (1)
994) 10, 286-292, Dalton S, and Treisman R (1992)
Characterization of SAP-1, aprotein recruited by s
erum response factor to the c-fos serum responseel
ement.Cell 68, 597-612, "MATCHMARKER Two-Hybrid
System '', `` MammalianMATCHMAKER Two-Hybrid Assay Ki
t '', `` MATCHMAKER One-Hybrid System '' (all manufactured by Clontech), `` HybriZAP Two-Hybrid Vector Syste
m "(Stratagene). In the two-hybrid system, the protein of the present invention or a partial peptide thereof is fused with an SRF DNA binding region or a GAL4 DNA binding region and expressed in yeast cells to express a protein that binds to the protein of the present invention. From the cells expected to produce a cDNA library that is expressed in a form fused with the VP16 or GAL4 transcriptional activation region, and introduced into the yeast cells, the library-derived cDNA was detected from the detected positive clones. Isolate (when the protein that binds to the protein of the present invention is expressed in yeast cells, the binding of both activates the reporter gene, and a positive clone can be confirmed). Isolated
By introducing the cDNA into E. coli and expressing it, the cDNA
Can be obtained. As a result, it is possible to prepare a protein that binds to the protein of the present invention or a gene thereof. Reporter genes used in the 2-hybrid system include:
For example, in addition to the HIS3 gene, Ade2 gene, LacZ gene, CA
T gene, luciferase gene, PAI-1 (Plasminogena
ctivator inhibitor type1) gene etc.
Not limited to these.

The screening for a compound that binds to the protein of the present invention can also be performed by using affinity chromatography. For example, the protein of the present invention is immobilized on a carrier of an affinity column, and a test sample which is expected to express a protein that binds to the protein of the present invention is applied thereto. In this case, examples of the test sample include a cell extract, a cell lysate, and the like. After applying the test sample, the column can be washed to prepare a protein bound to the protein of the present invention.

The obtained protein is analyzed for its amino acid sequence, an oligo DNA is synthesized based on the amino acid sequence, and a cDNA library is screened using the DNA as a probe to obtain a DNA encoding the protein. it can.

In the present invention, a biosensor utilizing the surface plasmon resonance phenomenon can be used as a means for detecting or measuring the bound compound. A biosensor utilizing the surface plasmon resonance phenomenon enables real-time observation of the interaction between the protein of the present invention and the test compound as a surface plasmon resonance signal using a trace amount of protein and without labeling. (Eg, BIAcore, manufactured by Pharmacia). Therefore, BI
By using a biosensor such as Acore, it is possible to evaluate the binding between the protein of the present invention and a test compound.

The method for isolating not only proteins but also compounds (including agonists and antagonists) that bind to the protein of the present invention includes, for example, immobilizing the protein of the present invention, a synthetic compound, and a natural product bank. Or a method of screening a molecule that binds to the protein of the present invention by using a random phage peptide display library, or a high-throughput screening method using combinatorial chemistry technology (Wrighton NC; Farrell FX; ChangR; Kas
hyap AK; Barbone FP; Mulcahy LS; Johnson DL; Barret
t RW; JolliffeLK; Dower WJ., Small peptides as pot
ent mimetics of the protein hormoneerythropoietin,
Science (UNITED STATES) Jul 26 1996, 273 p458-6
4, Verdine GL., The combinatorial chemistry of nat
ure.Nature (ENGLAND) Nov 7 1996, 384 p11-13, Hoga
n JC Jr., Directed combinatorial chemistry.Nature
(ENGLAND) Nov 7 1996, 384 p17-9) are known to those skilled in the art.

Further, the present invention relates to the protein of the present invention and PKCI
(Razin E. et al., J. Biol. Chem. Vol. 274, 34272-34
276 (1999)) or siva1 (Yoon Y. et al., Oncogene.
vol. 18, 7174-7179 (1999)). This compound screening method comprises the steps of: (a) contacting the protein of the present invention or a partial peptide thereof with PKCI or siva1 in the presence of a test sample; (b) detecting the binding thereof.
(C) a step of selecting a compound having an activity of promoting or inhibiting the binding thereof as compared with the case where the compound is detected in the absence of the test sample.

Each protein (or partial peptide thereof) used in these screenings is not particularly limited as long as it has a binding ability or a complex-forming ability. Further, the protein may be a protein having a natural amino acid sequence or a mutant. Further, it may be a naturally-derived protein or a recombinant protein. Further, it may be a fusion protein with another peptide. As the partial peptide of the protein of the present invention, a partial peptide containing positions 35 to 604 of SEQ ID NO: 14 can be suitably used. Also, PKCI or siva
For 1, a human-derived protein or a protein derived from another animal can be used. Mutants of siva1 used in the screening of the present invention include those described in the literature (Yoon Y. et al., Oncoge
ne. vol. 18, 7174-7179 (1999)) mutant of siva1 (siva1 having several tens of amino acids inserted and siva1 alte
Probably an rnative splice variant. The variant is designated herein as "siva3"). The test sample is not particularly limited, for example,
Cell culture supernatant, fermentation microorganism product, marine organism extract, plant extract, prokaryotic cell extract, eukaryotic single cell extract or animal cell extract or their library, purified or crude protein, peptide, Examples include peptidic compounds, synthetic low molecular weight compounds, and natural compounds. Proteins or peptides used in the screening of the present invention, for example,
The protein may be any of a purified protein, a soluble protein, a form bound to a carrier, a fusion protein with another protein, a form expressed on a cell membrane, and a form as a membrane fraction.

Such a screening can be performed, for example, using a two-hybrid method using yeast or animal cells. Specifically, a DNA encoding the protein of the present invention and a DNA encoding PKCI or siva1 were GAL4
Can be fused with any of the DNA-binding sequence and the sequence encoding the transcription activation domain, and co-expressed in a cell with a GAL4 luciferase reporter vector or the like, and the luciferase activity can be used as an index to screen for a promoter or inhibitor. .

[0121] Screening can also be performed using, for example, immunoprecipitation. In the presence of the test sample, the protein of the present invention is incubated with PKCI or siva1, and the complex is recovered using an antibody against one protein or peptide, or an antibody against a tag fused to the protein, and then the other. By detecting the protein using an antibody against the protein or the like, the protein of the present invention and PK
Binding to CI or siva1 can be evaluated. If the other protein is labeled with another tag or the like, detection can be facilitated. In addition, one protein is bound to a support, the other protein is bound, and a test sample is applied thereto. The effect of the test sample can be examined by detecting whether the bound protein is dissociated. Screening can also be performed using ELISA.

Screening using surface plasmon resonance as described in Screening for a protein binding to the protein of the present invention, or high-throughput screening using combinatorial chemistry can also be performed.

The compound isolated by the screening of the present invention becomes a candidate for a drug for promoting or inhibiting the activity of the protein of the present invention, and a disease caused by abnormal expression or function of the protein of the present invention or the present invention. Application to the treatment of diseases that can be treated by controlling the activity of the protein is considered. Substances obtained by using the screening method of the present invention and having a part of the structure of the compound having the activity of binding to the protein of the present invention, which are converted by addition, deletion and / or substitution, also bind to the protein of the present invention. Included in the compound.

The protein of the present invention or a compound which can be isolated by the screening of the present invention may be obtained from humans or mammals such as mice, rats, guinea pigs, rabbits, chickens, cats, dogs, sheep, pigs, cows, monkeys, baboons and chimpanzees. When used as a medicine, besides directly administering the protein or the isolated compound itself to the patient,
It is also possible to formulate and administer by a known pharmaceutical method. For example, tablets coated with sugar as needed,
It can be used orally as capsules, elixirs, microcapsules or parenterally in the form of injections of sterile solutions or suspensions in water or other pharmaceutically acceptable liquids. For example, a pharmacologically acceptable carrier or medium, specifically, sterile water or physiological saline, vegetable oil, emulsifier, suspending agent, surfactant, stabilizer, flavor,
It is conceivable to formulate the compound by appropriately combining it with excipients, vehicles, preservatives, binders and the like, and mixing it in a unit dose form required for generally accepted pharmaceutical practice. The amount of the active ingredient in these preparations is such that an appropriate dose in the specified range can be obtained.

Examples of additives that can be mixed with tablets and capsules include binders such as gelatin, corn starch, tragacanth gum, gum arabic, excipients such as crystalline cellulose, corn starch, gelatin,
Swelling agents such as alginic acid, lubricants such as magnesium stearate, sweeteners such as sucrose, lactose or saccharin, and flavoring agents such as peppermint, reddish oil or cherry are used. When the unit dosage form is a capsule, the above-mentioned materials may further contain a liquid carrier such as an oil or fat. Sterile compositions for injection can be formulated using vehicles such as distilled water for injection, according to standard formulation practice.

Examples of the aqueous solution for injection include physiological saline, isotonic solution containing glucose and other adjuvants, for example,
Sorbitol, D-mannose, D-mannitol, sodium chloride, and a suitable solubilizing agent such as an alcohol, specifically ethanol, polyalcohol such as propylene glycol, polyethylene glycol, a nonionic surfactant such as polysorbate 80 (TM), HC
You may use together with O-50.

The oily liquid includes sesame oil and soybean oil, and may be used in combination with benzyl benzoate or benzyl alcohol as a solubilizer. Also, buffers, such as phosphate buffers, sodium acetate buffers, soothing agents, such as
Procaine hydrochloride, stabilizers such as benzyl alcohol,
You may mix with phenol and an antioxidant. The prepared injection is usually filled into an appropriate ampoule.

The administration to a patient can be carried out, for example, by intraarterial injection, intravenous injection, subcutaneous injection and the like, or intranasally, transbronchially, intramuscularly, transdermally, or orally by a person skilled in the art. It can be done by a method. The dose varies depending on the weight and age of the patient, the administration method, and the like, but those skilled in the art can appropriately select an appropriate dose. In addition, the compound
As long as it can be encoded by DNA, it is also conceivable to integrate the DNA into a vector for gene therapy and perform gene therapy. The dose and administration method vary depending on the patient's weight, age, symptoms, and the like, but can be appropriately selected by those skilled in the art.

The dose of the protein of the present invention, the compound that binds to the protein of the present invention, or the compound that modulates the activity of the protein of the present invention varies depending on the condition. (Assuming a weight of 60 kg)
In about 0.1 to 100 mg per day, preferably about
It is believed to be between 1.0 and 50 mg, more preferably between about 1.0 and 20 mg.

In the case of parenteral administration, the single dose varies depending on the subject of administration, the target organ, the condition and the method of administration. It may be convenient to administer about 0.01 to 30 mg, preferably about 0.1 to 20 mg, more preferably about 0.1 to 10 mg per day by intravenous injection. In the case of other animals, the amount converted to 60 kg of body weight,
Alternatively, an amount converted per body surface area can be administered.

[0131]

EXAMPLES Hereinafter, the present invention will be described specifically with reference to Examples, but the present invention is not limited to these Examples.

[Example 1] Isolation of YS68 gene In order to obtain a molecule that is specifically expressed in hemangioblast,
From the yolk sac-derived cDNA on day 9 of the embryo to the cDNA of the yolk sac on the day 14 of the embryo
An experiment to subtract A was performed. Poly A RNA was purified from mouse yolk sac of embryonic day 9 and mouse yolk sac of embryonic day 14, and subtracted using PCR-Select cDNA Subtraction Kit (Clontech). The obtained cDNA fragment is pGEM
The DNA was subcloned into a -T vector (Promega), and a cDNA highly expressed in the yolk sac on the 9th day of the embryo was selected by dot blotting and sequenced. Clone # 68 was a novel gene fragment that was not registered in the database, so primers were further designed from the sequence of this gene fragment,
e Using the 15-day Embryo Marathon-Ready cDNA (Clontech) as a template, a full-length cDNA was isolated using the 5'-RACE method. Mouse YS68 encodes 1265 amino acids, but it is expected that a further upstream sequence exists.

The obtained YS68 had no characteristic motif in its amino acid sequence. However, the presence of multiple nuclear localization signals was confirmed. From this, YS68 was expected to be a protein that functions in the nucleus.
Therefore, in order to confirm this, a vector expressing a protein in which a Flag tag has been added to the mouse YS68 protein (1265 amino acids) [pEFBOSE-Flag (Nakashima et al. (1997) FE
BS Let. 403, 79-82)] was transfected into COS7 cells. 24 hours later, cells are fixed with 4% formalin and 0.1% Tr
Processed with iton-X 100. And anti-Flag antibody, followed by FITC
It was reacted with labeled anti-mouse IgG and observed with a fluorescence microscope.
As a result, YS68 was strongly expressed in the nucleus as expected (FIG. 1). The cell nucleus is where DNA transcription takes place, and therefore YS68 is expected to be a transcription factor involved in DNA transcription.

For the human YS68 gene, primers were designed based on the gene sequence of mouse YS68, and 5′-RACE and 3′-
It was isolated by the RACE method. That is, based on the mouse YS68 gene sequence, an EST fragment considered to be a human YS68 homolog was searched from the EST database. Primers are designed based on this EST fragment, and human fetal liver Marathon-Rea
Using the dy cDNA (CLONTECH) as a template, the 5'-region and 3'-region cDNA were converted to 5'-R according to the attached protocol.
It was isolated by the ACE and 3'-RACE methods. Isolated cDNA
Is shown in SEQ ID NO: 11, and the amino acid sequence of the protein encoded by the cDNA is shown in SEQ ID NO: 12.
FIG. 8 shows a comparison of the amino acid sequences of human and mouse YS68.

Example 2 Expression Pattern Analysis of YS68 The distribution of YS68 expression in tissues was analyzed by Northern blotting. ISOGEN from fetal or adult mouse tissues
(Wako Pure Chemical Industries) was used to prepare total RNA. these
After electrophoresis at 25 μg / lane and blotting on a nylon membrane, it was hybridized with a YS68 cDNA fragment labeled with ³² P. Hybridization is ExpressHyb
Perform at 68 ° C for 2 hours in a solution (Clontech)
Then, after washing several times at room temperature with 2 × SSC and 0.1% SDS,
After washing several times with SSC and 0.1% SDS at 65 ° C., autoradiography was performed.

The expression of YS68 was highest in testis of adult tissues, and then in kidney and lung. YS in hematopoietic tissue
Looking at the expression of 68, it was very strongly expressed in the liver, spleen, and thymus, which function as hematopoietic tissues during the fetal period. However, expression in these tissues in adults was rapidly reduced or eliminated (Figure 2).

Therefore, the expression pattern in tissues known to be involved in early hematopoiesis was examined in further detail.
From studies to date, it is known that the hematopoietic field shifts as follows during the embryonic period of the mouse. First, fetal hematopoiesis begins in the yolk sac on day 8 of the embryo, and later, adult hematopoiesis begins in the AGM region on day 10.5 of the embryo. Blood cells generated by AGM immediately migrate to the liver formed around 11.5 days of fetal life, where they differentiate and proliferate until immediately after birth. On the other hand, hematopoiesis will also be performed in the thymus and spleen around 16.5 days of embryo. After birth, the place of hematopoiesis turns into bone marrow. Based on this fact, the expression pattern of YS68 in these tissues was analyzed in detail by RT-PCR. Total RNA was extracted from each tissue of mouse embryo or adult mouse at each stage.
μg of total RNA to SUPERSCRIPT II preamplification
Reverse transcription was performed on cDNA using a system (manufactured by Gibco). Using this as a template, a primer specific to YS68 (68-3: 5'-
CACCCGTGAAGAAACAAATAGGCA-3 '/ SEQ ID NO: 3, 68.4: 5'
-CCTTTGGTACATGAGCTTCTATTT-5 '/ SEQ ID NO: 4) or PCR reaction with primers specific to G3PDH (94 ° C for 30 seconds,
25 cycles of 30 seconds at 62 ° C. and 30 seconds at 72 ° C.), electrophoresed on a 1% agarose gel, and stained with ethidium bromide.

First, in the yolk sac, as the development progresses, YS68
Expression gradually decreased (Fig. 3). YS68 expression was unexpectedly low in the AGM region of embryonic day 10.5 at which adult hematopoiesis begins (FIG. 5A). On the other hand, in the liver, thymus, and spleen, which are known to function as hematopoietic fields during the fetal period, YS68 expression was extremely high in correlation with the period of functioning as a hematopoietic organ (FIG. 4).

Next, the expression distribution of YS68 in mouse embryos was
Analyzed by in situ hybridization. YS
545 bp cDNA (positions 898 to 1443) of 68 5 'region
Using the vector created by inserting into criptII as a template, T7R
In vitro transcription was performed using NA polymerase or T3 RNA polymerase (Boehringer Mannheim) to synthesize sense or antisense ³⁵ S-labeled RNA, respectively. The mouse fetus was removed, frozen, and sectioned using a cryostat. After fixation with 4% paraformaldehyde / PBT and acetylation, hybridization was carried out at 55 ° C. overnight with the above RNA probe. this
After treatment with RNase A, the cells were washed several times and subjected to autoradiography.

YS68 was most strongly expressed in the liver at 11.5 days of embryonic age (FIG. 6). On embryonic day 14.5, it was strongly expressed mainly in the liver and the developing thymus, and was also observed in the lung and neural tube (FIG. 7).

These results indicate that the expression of YS68 is restricted to tissues in which hematopoiesis is actively performed in a time-specific manner, indicating that YS68 is a molecule involved in early hematopoiesis. It is strongly suggested that Expression was low in the AGM region at embryonic day 10.5, which is thought to be the site of hematopoietic cell development, but the absolute number of hematopoietic cells was AG
This may be because the whole M area is not so high. In fact, Suda et al.'S group used TEK as a marker for hemangioblasts and showed that the proportion of hemangioblasts in the AGM region at embryonic day 10.5 was less than 5% (Hamaguchi, I. et. al., (1999) Blood, 93, 154
9-1556). On the other hand, when the AGM region of embryonic day 10.5 is dispersed and cultured in a petri dish, the appearance of blood cells is observed in about 5 days of culture (Mukouyama, Y. et al., (1998) Immunit
y, 8, 105-114). Interestingly, AGM cultured for 5 days
In derived cells, the expression of YS68 was increased (FIG. 4B).
From these results, it is expected that the expression of YS68 will increase in cells that have acquired hematopoietic activity or in immature blood cells.

[Example 3] Full-length cloning of mouse and human YS68 Mouse 15-day Embryo Marathon-Rea was prepared using primers designed from the YS68 gene sequence obtained so far.
dy cDNA and human fetal liver Marathon Ready
Using the cDNA (CLONTECH) as a template, 5'-race method was performed to clone the 5 'upstream region of mouse and human YS68 gene. By repeating this 5'-RACE, the sequences of human and mouse full-length cDNAs were determined.

As a result, it was predicted that human YS68 encoded 2266 amino acids and mouse YS68 encoded 2243 amino acids (FIG. 9). As a result of comparing the amino acid sequences of human and mouse, it is interesting that the N-terminal region (human 1-1137, mouse 1-11
37) had a very high homology of 87%, but 57% in the central region (human 1138-1683, mouse 1138-1679) and the C-terminal region (human 1684-2266, mouse 1680-2243) Then 4
It showed a very low homology of 5%. Several nuclear localization signals were present in the C-terminal region with low homology. On the other hand, in the highly homologous N-terminal region, there were two WD repeats known to be necessary for the interaction between proteins. The very high homology in this region in humans and mice suggests that this region may be important in the function of YS68.

[Example 4] Protein binding to YS68 Since a protein binding site (WD repeat) in the N-terminal region exists, it was considered that YS68 may be bound to any protein in vivo. Therefore, cell culture cells from fetal liver
Lysate was prepared, immunoprecipitated with an anti-YS68 antibody, and subjected to SDS polyacrylamide gel electrophoresis to examine whether a protein co-precipitating with YS68 was present. Specifically, cultured mouse liver cells at 14.5 days of embryonic embryo were lysed in lysis buffer (lysate).
sis buffer; 0.5% NP-40, 10mM Tris-HCl pH7.6, 150mM
NaCl, 5mM EDTA, 2mM Na ₃ VO ₄ , 1mM phenylmethylsulfonyl fluorid
e), 5 μg / ml aprotinin). After incubating overnight at 4 degrees with anti-YS68 antibody, protein G was added and incubated for another hour. The immunoprecipitate was electrophoresed on an SDS polyacrylamide gel,
Silver staining was performed.

As a result, it was confirmed that a plurality of molecules co-precipitated with YS68 were present in fetal liver cells (FIG. 10). This suggested that YS68 functions by binding to several types of proteins in cells.

Example 5 Expression Site of YS68 in Tissue In order to analyze the expression site of YS68 in more detail, rabbits were immunized with YS68 protein to prepare a polyclonal antibody against YS68. In preparing a YS68 polyclonal antibody, a protein encoding the 1208-1582 amino acid region of mouse YS68 was expressed in Escherichia coli, purified according to a standard method, and used as an antigen. A rabbit (New Zealand white, 2.5 kg, female) was immunized four times with 200 ug every 10 days, and the whole blood was collected to obtain an antiserum. In addition, an affinity column with immobilized antigen was prepared, and anti-YS68
The polyclonal antibody was purified.

Using this antibody, the expression site in the AGM region of a fetal 11.5 day fetus was examined by immunostaining. For tissue staining, a section was first prepared from a frozen mouse embryo using a cryostat (Lyca). This was fixed with 4% formaldehyde and treated with methanol. After treatment with 0.3% hydrogen peroxide solution,
Perform blocking with 3% BSA and incubate primary antibody at 4 ° C overnight.
After reacting with a secondary antibody (HRP-labeled anti-rabbit IgG) at room temperature for 1 hour, washing with PBS three times, the substrate (Metal Enhanced DAB Su
bstrate kit, PIERCE) was added for color development.

As a result, the blood cells present in the endothelium were stained by staining with the erythrocyte marker TER119 used as a control (FIGS. 11A and 11B), whereas the vascular endothelium was stained specifically by staining with YS68. (FIGS. 11C, D, E). Interestingly, YS68 was also strongly stained in the blood cells emerging from the endothelium (FIG. 11E, arrow). YS68 was also strongly expressed on the vascular endothelium of the umbilical artery (FIG. 11F). Embryo 9.
Even in the yolk sac blood vessels on day 5, TER119 specifically stained the blood cells in the blood vessels, whereas YS68 was strongly expressed in the vascular endothelium rather than the blood cells (Fig. 11G, H).

[Example 6] Expression of YS68 in cells [0149] Liver was surgically excised from a fetus (embryo 14.5 days), finely chopped with tweezers, and subjected to cell disociation buffe.
Incubated in r (GIBCO) at 37 degrees for 30 minutes. This was further treated with 0.1% collagenase at 37 ° C. for 1 hour, and then the cells were loosened by pipetting. After washing the cells several times with PBS, the cells were suspended in DMEM containing 10% FCS and cultured in a 10 cm dish.

To examine the localization of endogenous YS68 in cells, cultured hepatic cells were stained with YS68. For cell staining, first fix cells in 4% formalin and
Treated with 1% Triton-X 100. Then, it was reacted with the primary antibody and subsequently with the secondary antibody, and the color was developed as in Example 5.

As a result, it was found that YS68 had a plurality of nuclear translocation signals, but was strongly expressed not only in the nucleus but also in some cells around the nucleus (FIG. 12). Next, expression in blood cells was similarly analyzed. It was found that the expression level of YS68 in blood cells separated from fetal liver varied depending on the cell type (FIG. 13B).

Thus, the blood cell population was sorted by CD34 which is a marker of immature blood cells, and the expression of YS68 in CD34-positive cells was examined. For collection of CD34-positive cells, first, fetal liver (embryonic day 14.5) was incubated at 37 ° C. for 30 minutes with a dissociation buffer, and then separated by pipetting in PBS.
After passing through a nylon mesh filter (FALCON), the suspension was suspended in a sample buffer (0.5% BSA, 2 mM EDTA in PBS). This was reacted with biotin-labeled anti-CD34 antibody (Pharmingen), followed by FITC-labeled streptavidin at 4 degrees, and then incubated with anti-FITC microbeads, MACS
Using a (Magnetic Cell Sorting) column, CD34-positive cells were eluted according to the attached protocol. The cells were centrifuged at 400 rpm for 5 minutes on a slide glass and fixed on the slide glass. Cell staining was performed as described above.

As a result, blood cells enriched with CD34 (FIG. 12D)
YS6 compared to blood cells that passed through the CD34 column (Figure 12C).
The expression of 8 was found to be high. This suggested that YS68 was mainly expressed in CD34-positive and more undifferentiated blood cells in blood cells.

[Example 7] Localization of each domain of YS68 in cells Using cDNA prepared from mouse fetal liver as a template, mouse YS
CDNAs encoding the 68 N-terminal region (amino acids 5-1148) and the C-terminal region (amino acids 981-2243) were amplified by PCR.
This is inserted downstream of the Flag region of the animal cell expression vector pEFBOSE-F, and expresses the N-terminal region of YS68 in pEFBOSE-F-YS68.
(5-1148), and pEFBOSE-FY expressing the C-terminal region of YS68
S68 (981-2243) was produced. This was transfected into COS-7 cells using Lipofectamine 2000 (GIBCO).
Four hours later, the cells were fixed with methanol. Then, in order to examine the localization of each expressed YS68 in the cells, the cells were anti-fl
The antibody was reacted with an ag antibody, followed by a peroxidase-labeled anti-mouse IgG, and finally a substrate was added for color development.

[0155] YS68 has a plurality of nuclear localization signals in the C-terminal region (Fig. 9). From this, localization in the nucleus was predicted. In contrast, endogenous YS68 is not only nuclear but also nuclear. (Fig. 12). Therefore, a construct lacking the N-terminal region or the C-terminal region of YS68 was prepared, expressed in COS cells, and examined for its localization. As a result, YS68 lacking the C-terminal region was strongly localized in the cytoplasm, and YS68 lacking the N-terminal region was strongly localized in the nucleus (FIG. 14). These results suggested that the N-terminal region may inhibit the transfer of YS68 to the nucleus. Since there are two WD repeats required for protein interaction in the N-terminal region, it was speculated that the translocation to the nucleus might be inhibited by binding of this region to any molecule.

[Example 8] Analysis of transcription activation domain of YS68 From the experiments thus far, localization of YS68 was recognized in the nucleus and cytoplasm, and YS68 from which the N-terminal region was deleted was strongly expressed in the nucleus. It turned out to be localized. This suggested that YS68 may function as a transcription factor in cells. Therefore, the transcription activation domain of YS68 was analyzed using a yeast system.

First, PCR was performed using human fetal liver cDNA (CLONTECH) as a template, and YS68 derived from human was divided into three major regions (amino acids, 35-604, 583-1134, 1080-1630;
A gene fragment encoding FIG. 15A) was obtained. Each of these gene fragments was inserted into pODB80, and a fusion protein expression vector with the GAL4-DNA binding domain (pODB-35-604, pODB-583-113)
4, pODB-1080-1630). After transforming each of these vectors into an adenine- and histidine-requiring yeast yeast strain AH109 according to a standard method, each colony was wound on a plate lacking adenine and histidine and adding X-?-Gal. One week later, the colonies that grew were observed. The adenine, histidine and lacZ genes are located downstream of the GAL4 promoter, so if the expressed protein has transcriptional activity, adenine and histidine will be deleted and a blue colony will be formed on the plate supplemented with X-?-Gal. be able to.

As a result, as shown in FIG.
It was confirmed that only the yeast expressing the 1630 region formed a blue colony and grew. From this, YS68
It was revealed that a transcription activation region exists in 1080-1630.

Example 9 Identification of YS68 Binding Protein As shown in FIG. 10, YS68 was expected to bind to multiple proteins in cells. Thus, yeast two-hybrid screening was performed to identify molecules that bind to YS68.

Yeast 2-hybrid screening includes YS
The two N-terminal regions of 68 (35-604, 583-1134) were
(PODB-35-604, pODB-583-1134) was used. Yeast transformed with these vectors (AH109
Strain) has a human fetal liver with a GAL4 activation domain
A matchmaker cDNA library (CLONTECH) was introduced, and these were wrapped around adenine, histidine, tryptophan, and leucine deletion plates. One week later, colonies that had grown on the plate were collected. From the growing colonies, cDNA fragments were increased by PCR, and their nucleotide sequences were determined. To further confirm this reproducibility, these gene fragments were again inserted downstream of the GAL4 activation domain of pACT2, and introduced into a yeast strain transformed with pODB-35-604 and pODB-583-1134. After growing this on a plate containing adenine and histidine, this colony was wound on a plate lacking adenine and histidine and added with X-?-Gal, and it was confirmed that the colony could be grown on this plate.

As a result, protein kinase C inhibitor 1
(PKCI), two ESTs, and CD27 binding protein (siv
a1, siva3) was found to bind to the region 35-604.
In addition, these proteins do not bind to the 583-1134 region,
It was found that it specifically binds to the 04 region (Fig. 1
6).

[0162]

According to the present invention, a novel protein "YS68" predicted to be involved in early hematopoiesis and a gene encoding the protein are provided. The gene can be used as a marker for hematopoietic cells involved in early hematopoiesis or as a factor for controlling hematopoiesis. In addition, it can be used as a tool for the purification and cloning of new factors involved in hematopoiesis, and also for drug development for various diseases caused by abnormal expression of the gene of the present invention due to abnormal regulation of expression in vivo. It is. The “YS68” gene of the present invention is also considered to be involved in hematological tumors. Therefore, drug development for tumors utilizing the protein of the present invention is expected. By designing a drug or the like targeting the gene of the present invention, it is considered that a drug with a new mechanism of action can be developed. In particular, human-derived proteins and genes are more suitable for drug development and the like than those derived from other organisms.

[0163]

[Sequence List] SEQUENCE LISTING <110> CHUGAI RESEARCH INSTITUTE FOR MOLECULER MEDICINE, INC. <120> YS68, a novel gene involved in early hematopoiesis <130> C2-112DP2 <140> <141> <150> JP 11-288738 <151> 1999-10-08 <150> JP 11-288739 <151> 1999-10-08 <150> JP 2000-123721 <151> 2000-04-19 <150> PCT / JP00 / 05756 <151> 2000-8-25 <160> 15 <170> PatentIn Ver. 2.0 <210> 1 <211> 4115 <212> DNA <213> Mus musculus <220> <221> CDS <222> (1) .. (3817) <400> 1 t cag att ctg aag aat aat ctc atg agt gat cgt gac cct cga ttg cgg 49 Gln Ile Leu Lys Asn Asn Leu Met Ser Asp Arg Asp Pro Arg Leu Arg 1 5 10 15 gaa aga tcg gtg act cga aat tct ata tta gac cag tat ggg aaa atc 97 Glu Arg Ser Val Thr Arg Asn Ser Ile Leu Asp Gln Tyr Gly Lys Ile 20 25 30 cta cct aga gtc cag aga aag tta gct gtt gag cga gct aag cct tac 145 Leu Pro Arg Val Gln Arg Lys Leu Ala Val Glu Arg Ala Lys Pro Tyr 35 40 45 cac ctg tcg aca tcc tca gtt ttt cat gaa gtt tct aga ccc aaa ccg 193 His Leu Ser Thr Ser Ser Val Phe His Glu Val Ser Arg Pro Lys Pro 50 55 60 tta tcg gca ttt cca aag aaa gct ata act gga aca gtg tta acc cga 241 Leu Ser Ala Phe Pro Lys Lys Ala Ile Thr Gly Thr Val Leu Thr Arg 65 70 75 80 tct acg ttc atc agc aat gtt tta tct aaa att gga gag gtg tgg gca 289 Ser Thr Phe Ile Ser Asn Val Leu Ser Lys Ile Gly Glu Val Trp Ala 85 90 95 agt cat gag cct aga aat ggc gtc tca ctt ttt aac agt cct aaa aca 337 Ser His Glu Pro Arg Asn Gly Val Ser Leu Phe Asn Ser Pro Lys Thr 100 105 110 gaa cag cca tct cct gta gta cac tct ttc cca cac cca gag ctt cct 385 Glu Gln Pro Ser Pro Val Val His Ser Phe Pro His Pro Glu Leu Pro 115 120 125 gag gcg ttt gtt gga act cca att tca aat aca tcc cag aga att tct 433 Glu Ala Phe Val Gly Thr Pro Ile Ser Asn Thr Ser Gln Arg Ile Ser 130 135 140 aga tta ctg gat ttg gtt gtc cat cct gta ccc cag cct tct cag tgt 481 Arg Leu Leu Asp Leu Val Val His Pro Val Pro Gln Pro Ser Gln Cys 145 150 155 160 ttg gag ttt att caa caa agt ccc aca aga tct cct ttg tgt ctg ctg 529 Leu Glu Phe Ile Gln Gln Ser Pro Thr Arg Ser Pro Leu Cys Leu Leu 165 170 175 tcc agt tcg tta cca tta agt tca cag ttt aaa agg cca cat cag aat 577 Ser Ser Ser Leu Pro Leu Ser Ser Gln Phe Lys Arg Pro His Gln Asn 180 185 190 acc tcc agg cct tca gag ttg ctt tta ctt gag act cct ctc ata gtt 625 Thr Ser Arg Pro Ser Glu Leu Leu Leu Leu Glu Thr Pro Leu Ile Val 195 200 205 aag aaa gct aaa tct ttg gct ctg tca gcc acg tct tct gga ttt gcc 673 Lys Lys Ala Lys Ser Leu Ala Leu Ser Ala Thr Ser S er Gly Phe Ala 210 215 220 gag ttt act cct cca tcc atc ctt agg tct ggt ttt cga aca aca cct 721 Glu Phe Thr Pro Pro Ser Ile Leu Arg Ser Gly Phe Arg Thr Thr Pro 225 230 235 240 tta gca tct ccc tct ttg tca cct gga aga tct ctc act ccg cct ttc 769 Leu Ala Ser Pro Ser Leu Ser Pro Gly Arg Ser Leu Thr Pro Pro Phe 245 250 255 aga gtt aaa gaa aca agg att tca ttc atg gaa gaa ggc atg aat aca 817 Arg Val Lys Glu Thr Arg Ile Ser Phe Met Glu Glu Gly Met Asn Thr 260 265 270 cac tgg act gat aga gct aca gat gac cga aat aca aaa gcg ttt gtt 865 His Trp Thr Asp Arg Ala Thr Asp Asp Arg Asn Thr Lys Ala Phe Val 275 280 285 agc aca tct ttc cat aaa tgt gga ctt cca gca gaa act gag tgg atg 913 Ser Thr Ser Phe His Lys Cys Gly Leu Pro Ala Glu Thr Glu Trp Met 290 295 300 aag acc agt gat aag aat aca tat ttt cct ctg gat gtc cct gca aag 961 Lys Thr Ser Asp Lys Asn Thr Tyr Phe Pro Leu Asp Val Pro Ala Lys 305 310 315 320 ggc cct cag aaa gtg gtg gca gag tca ctg gct acc cat tca gga agg 1009 Gly Pro Gln Lys Val Val A la Glu Ser Leu Ala Thr His Ser Gly Arg 325 330 335 ctg gag aaa ctg gat gtg agc aaa gaa gac agc aca gct tcc acc agg 1057 Leu Glu Lys Leu Asp Val Ser Lys Glu Asp Ser Thr Ala Ser Thr Arg 340 345 350 tca gac cag acc tcc tta gag tat cat gac gca cca tca cca gaa gac 1105 Ser Asp Gln Thr Ser Leu Glu Tyr His Asp Ala Pro Ser Pro Glu Asp 355 360 365 ttg gaa ggt gct gtt ttt gtg tct ccc aag cca gca tct tcc tcc act 1153 Leu Glu Gly Ala Val Phe Val Ser Pro Lys Pro Ala Ser Ser Ser Thr 370 375 380 gaa cta act act aat tca act cta caa aca gag agg gat aat gat aaa 1201 Glu Leu Thr Thr Asn Ser Thr Leu Gln Thr Glu Arg Asp Asn Asp Lys 385 390 395 400 gat gcg ttt aag tca gaa ggt act cct tca ccc gtg aag aaa caa ata 1249 Asp Ala Phe Lys Ser Glu Gly Gly Thr Pro Ser Pro Val Lys Lys Gln Ile 405 410 415 ggc acg gga gac gct gca gtg gaa gca ttt tca gaa ctg agt cgc tta 1297 Gly Thr Gly Asp Ala Ala Val Glu Ala Phe Ser Glu Leu Ser Arg Leu 420 425 430 gac cct gtt gaa aga gct gaa gct tct ttt ggt gtg tcg tca gtc tgt Asp Pro Val Glu Arg Ala Glu Ala Ser Phe Gly Val Ser Ser Val Cys 435 440 445 gaa ggg gaa acc tcc act tca aac tcc aag acg tca gtt ctg gat gga 1393 Glu Gly Glu Thr Ser Thr Ser Asn Ser Lys Thr Ser Val Leu Asp Gly 450 455 460 atc gtg cct att gag agc cga acc tcc ata ctt aca gca gac cac aaa 1441 Ile Val Pro Ile Glu Ser Arg Thr Ser Ile Leu Thr Ala Asp His Lys 465 470 475 480 480 gag tct gtg gcc aac acg gtt gca gat gtt gaa agc tct ggg tcc acc 1489 Glu Ser Val Ala Asn Thr Val Ala Asp Val Glu Ser Ser Gly Ser Thr 485 490 495 agc tcc aag tgc ccg gtt acc tct gaa cgc agc ctc ggc caa aaa cta 1537 Ser Ser Lys Cys Pro Val Thr Ser Glu Arg Ser Leu Gly Gln Lys Leu 500 505 510 aca tta aac tta aaa gaa gat gaa ata gaa gct cat gta cca aag gag 1585 Thr Leu Asn Leu Lys Glu Asp Glu Ile Glu Ala His Val Pro Lys Glu 515 520 525 aac gtt ggt tta cca gaa gaa agc cct cga att tct gct gct cct tct 1633 Asn Val Gly Leu Pro Glu Glu Ser Pro Arg Ile Ser Ala Ala Pro Ser 530 535 540 gat act cac gag att cat cta att gga tgt gaa aat ctt gaa gtt caa 1681 Asp Thr His Glu Ile His Leu Ile Gly Cys Glu Asn Leu Glu Val Gln 545 550 555 560 aat tca gaa gag gag gcc aag aat ctt tca ttt gat gag ttg tat ccc 1729 Asn Ser Glu Glu Glu Ala Ly Asn Leu Ser Phe Asp Glu Leu Tyr Pro 565 570 575 tta ggg gca gag aaa ctt gag tat aat ctc agt act att gag cag cag 1777 Leu Gly Ala Glu Lys Leu Glu Tyr Asn Leu Ser Thr Ile Glu Gln Gln 580 585 590 ttt tgt gac ttg cct gat gac aaa gac tct gct gaa tgt gat gct gct 1825 Phe Cys Asp Leu Pro Asp Asp Lys Asp Ser Ala Glu Cys Asp Ala Ala 595 600 605 gaa gta gac ggg gaa ctt ttt gtg gcc cag agc aact acc 1873 Glu Val Asp Gly Glu Leu Phe Val Ala Gln Ser Asn Phe Thr Leu Ile 610 615 620 tta gaa ggt gaa gaa gga gaa gct gag gca agc gac tct gca gca cct 1921 Leu Glu Gly Glu Glu Gly Glu Ala Glu Ala Ser Ala Ala Pro 625 630 635 640 aat atg tta ccg aaa tcg acc aag gaa aaa cct gtg tgc tac agg gaa 1969 Asn Met Leu Pro Lys Ser Thr Lys Glu Lys Pro Val Cys Tyr Arg Glu 645 650 655 cc ccc cat aat c ag gag cgc gtt aca gat ttg cca tct gct gtg act gct 2017 Pro His Asn Gln Glu Arg Val Thr Asp Leu Pro Ser Ala Val Thr Ala 660 665 670 gac caa gaa tcc cac aag gta gag act tta ccg tat gtg cct gaa ccg 2065 Asp Gln Glu Ser His Lys Val Glu Thr Leu Pro Tyr Val Pro Glu Pro 675 680 685 gtt aaa gtg gca att gca gaa aat ctg ttg gat gta att aaa gac acc 2113 Val Lys Val Ala Ile Ala Glu Asn Leu Leu Asp Val Ile Lys Asp Thr 690 695 700 aga agt aag gaa gca act ccc gtg gca gca ggt gag gct ggt gat gag 2161 Arg Ser Lys Glu Ala Thr Pro Val Ala Ala Gly Glu Ala Gly Asp Glu 705 710 710 715 720 gac gga gca gtg ata gtc tca aag gct gca cat tcg tcc agg ctg aca 2209 Asp Gly Ala Val Ile Val Ser Lys Ala Ala His Ser Ser Arg Leu Thr 725 730 735 aac tct aca ccg aag act gtt aag gaa cca cgt gca gag act gta aat 2257 Asn Ser Thr Pro Lys Thr Val Lys Glu Pro Arg Ala Glu Thr Val Asn 740 745 750 acc agc cag agt gat gac atg gtt tct tct aga act ctc aca aga agg 2305 Thr Ser Gln Ser Asp Asp Met Val Ser Ser Arg Thr Leu Thr Arg Arg 755 760 765 cag cat gcc cta agc ctg aat gtc aca tca gaa caa gag cct tca gca 2353 Gln His Ala Leu Ser Leu Asn Val Thr Ser Glu Gln Glu Pro Ser Ala 770 775 780 gtt gcc act cct aag aag aga act aga aaa att aaa gaa act cct gag 2401 Val Ala Thr Pro Lys Lys Arg Thr Arg Lys Ile Lys Glu Thr Pro Glu 785 790 795 800 tct tct gaa agg acc tgt tct gac cta aaa gta gca cct gag aac caa 2449 Ser Ser Glu Arg Thr Cys Ser Asp Leu Lys Val Ala Pro Glu Asn Gln 805 810 815 ctg aca gct cag aat cct ccc gct cct agg aga aga aag aag aag gac 2497 Leu Thr Ala Gln Asn Pro Pro Ala Pro Arg Arg Arg Lys Lys Lys Asp 820 825 830 gtt agc caa ggc aca ctg cca agt tct ggt gct gtg gag ccg gag ccg 2545 Val Ser Gln Gly Thr Leu Pro Ser Ser Gly Ala Val Glu Pro Glu Pro 835 840 845 gaa cct cag ggt acg ccg gga aga ctg agg ctg aga acg cag cag 2593 Glu Pro Gln Gly Thr Pro Gly Arg Leu Arg Leu Arg Thr Gln Pro Pro 850 855 860 gag cca gca gct gaa gaa act cct tct aga aca aa aaa gtc agg ctt tca 2641 Glu Pro Ala Ala Glu Glu Thr Pro Ser Ar g Thr Lys Val Arg Leu Ser 865 870 875 880 tct gtt aga aag gga acc cct aga aga ctt aag aag tct gta gaa aat 2689 Ser Val Arg Lys Gly Thr Pro Arg Arg Leu Lys Lys Ser Val Glu Asn 885 890 895 895 ggg caa agt ata gaa att cta gat gat ctc aaa ggg agt gag gca gca 2737 Gly Gln Ser Ile Glu Ile Leu Asp Asp Leu Lys Gly Ser Glu Ala Ala 900 905 910 agt cat gac ggg act gtc aca gag ctg agg aat gcc aat 785 gaat Ser His Asp Gly Thr Val Thr Glu Leu Arg Asn Ala Asn Leu Glu Asp 915 920 925 act cag aat atg gag tat aaa caa gat gaa cac agt gac cag caa ccg 2833 Thr Gln Asn Met Glu Tyr Lys Gln Asp Glu His Ser Asp Gln Gln Pro 930 935 940 cct cta aaa cga aag agg gtc aga gag aga gaa gtt agt gtg tca agt 2881 Pro Leu Lys Arg Lys Arg Val Arg Glu Arg Glu Val Ser Val Ser Ser 945 950 955 960 gtg aca gaa gag cca aag ctt gac tca tcc cag ttg cct ctt cag aca 2929 Val Thr Glu Glu Pro Lys Leu Asp Ser Ser Gln Leu Pro Leu Gln Thr 965 970 975 gga ctc gat gta cct gcc acc cct agg aaa cgt ggt aga ccc agg aag 2977 Gly Leu A sp Val Pro Ala Thr Pro Arg Lys Arg Gly Arg Pro Arg Lys 980 985 990 gta gtt ccc tta gaa gct gac ggt ggc aca act ggt aag gaa cag aca 3025 Val Val Pro Leu Glu Ala Asp Gly Gly Thr Thr Gly Lys Glu Gln Thr 995 1000 1005 agt cct cag aag aaa gat gtt ccg gtt gtc cgg aga tct aca cgg aac 3073 Ser Pro Gln Lys Lys Asp Val Pro Val Val Arg Arg Ser Thr Arg Asn 1010 1015 1020 acc cca gct aga aat gtg agt act tta naa aaa tca gtt tta gtg cca 3121 Thr Pro Ala Arg Asn Val Ser Thr Leu Xaa Lys Ser Val Leu Val Pro 1025 1030 1035 1040 aat aag aa ga gct gct cta gtg gtg aca tct aag agg aga cct aca aag 3169 Asn Lys Glu Ala Ala Leu Val Val Thr Ser Lys Arg Arg Pro Thr Lys 1045 1050 1055 aag tct gca gag gaa agc tca aaa gat cca tca gcg gca gtc tca gac 3217 Lys Ser Ala Glu Glu Ser Ser Lys Asp Pro Ser Ala Ala Val Ser Asp 1060 1065 1070 tgg gcg ggt gga gca gcc cac aca gag tcc gct gac cga agg gac gga 3265 Trp Ala Gly Gly Ala Ala His Thr Glu Ser Ala Asp Arg Arg Asp Gly 1075 1080 1085 ctg ctt gcc gcc gct gct ctc acg cc a tct gcc cag ggc aca agg act 3313 Leu Leu Ala Ala Ala Ala Leu Thr Pro Ser Ala Gln Gly Thr Arg Thr 1090 1095 1100 agg tct aga agg acc atg ttg ttg acg gac att tct gaa ccc aaa act 3361 Arg Ser Arg Arg Thrg Met Leu Leu Thr Asp Ile Ser Glu Pro Lys Thr 1105 1110 1115 1120 gag cct tta ttt cct cct cct tca gtg aag gtt cca aag aaa aaa tca 3409 Glu Pro Leu Phe Pro Pro Ser Val Lys Val Pro Lys Lys Lys Ser 1125 1130 1135 aaa gct gag aac atg gag gcc gca gcc cag ctg aaa gaa ttg gtg tca 3457 Lys Ala Glu Asn Met Glu Ala Ala Ala Gln Leu Lys Glu Leu Val Ser 1140 1145 1150 gat tta tct tct cag ttt gtt gtt tcc cct cct aga acc agg 3505 Asp Leu Ser Ser Gln Phe Val Val Ser Pro Pro Ala Leu Arg Thr Arg 1155 1160 1165 cag aaa agt ata tcc aat act tcc aag ctt cta ggt gaa ctg gag agt 3553 Gln Lys Ser Ile Ser Asn Thr Ser Lys Leu Leu Gly Glu Leu Glu Ser 1170 1175 1180 gac cct aaa cca tta gag atc ata gaa caa aaa cca aaa aga agc agg 3601 Asp Pro Lys Pro Leu Glu Ile Ile Glu Gln Lys Pro Lys Arg Ser Arg 11 85 1190 1195 1200 act gtg aag aca aga gca agc aga aac aca gga aaa gga agt tct tgg 3649 Thr Val Lys Thr Arg Ala Ser Arg Asn Thr Gly Lys Gly Ser Ser Trp 1205 1210 1215 tca cct cct cct gta gaa att aag ctg gtt tct ccc ttg gcg agt cca 3697 Ser Pro Pro Pro Val Glu Ile Lys Leu Val Ser Pro Leu Ala Ser Pro 1220 1225 1230 gtg gat gaa ata aag acc ggc aag cca aga aaa act gca gaa ata gca 3745 Val Asp Glu Ile Lys Thr Gly Lys Pro Arg Lys Thr Ala Glu Ile Ala 1235 1240 1245 gga aaa act ctt gga agg ggc aga aag aag cca tct tct ttt cca aag 3793 Gly Lys Thr Leu Gly Arg Gly Arg Lys Lys Lys Pro Ser Ser Phe Pro Lys 1250 1255 1260 caa att tta cgc agg aaa atg ctg taatttttag cccaagattt taacacgcac 3847 Gln Ile Leu Arg Arg Lys Met Leu 1265 1270 ctgtttgtaa aagtcaacag tatttgtgtg gattattaaa gtcaccaatt tggatgaaaa 3907 tactttatat aaattgtaca attttgtaag cagtaaatga gtaactccac atggagtgca 3967 gttcttgtag tgcaggcgtt ttatacgact tgatgcgttt atatcaatgt aaatatgact 4027 tatcattggg aggttaaata aactactgta aagtaaaaaa aaaaaaaaaa a aaaaaaaaa 4087 aaaaaaaaaa aaaaaaaaaa aaaaaaaa 4115 <210> 2 <211> 1272 <212> PRT <213> Mus musculus <400> 2 Gln Ile Leu Lys Asn Asn Leu Met Ser Asp Arg Asp Pro Arg Leu Arg 1 5 10 15 Glu Arg Ser Val Thr Arg Asn Ser Ile Leu Asp Gln Tyr Gly Lys Ile 20 25 30 Leu Pro Arg Val Gln Arg Lys Leu Ala Val Glu Arg Ala Lys Pro Tyr 35 40 45 His Leu Ser Thr Ser Ser Val Phe His Glu Val Ser Arg Pro Lys Pro 50 55 60 Leu Ser Ala Phe Pro Lys Lys Ala Ile Thr Gly Thr Val Leu Thr Arg 65 70 75 80 Ser Thr Phe Ile Ser Asn Val Leu Ser Lys Ile Gly Glu Val Trp Ala 85 90 95 Ser His Glu Pro Arg Asn Gly Val Ser Leu Phe Asn Ser Pro Lys Thr 100 105 110 Glu Gln Pro Ser Pro Val Val His Ser Phe Pro His Pro Glu Leu Pro 115 120 125 Glu Ala Phe Val Gly Thr Pro Ile Ser Asn Thr Ser Gln Arg Ile Ser 130 135 140 Arg Leu Leu Asp Leu Val Val His Pro Val Pro Gln Pro Ser Gln Cys 145 150 155 160 Leu Glu Phe Ile Gln Gln Ser Pro Thr Arg Ser Pro Leu Cys Leu Leu 165 170 175 Ser Ser Ser Leu Pro Leu Ser Ser Gln Phe Lys Arg Pro His Gln Asn 180 185 190 Thr Ser Arg Pro Ser Glu Leu Leu Leu Leu Glu Thr Pro Leu Ile Val 195 200 205 L ys Lys Ala Lys Ser Leu Ala Leu Ser Ala Thr Ser Ser Gly Phe Ala 210 215 220 Glu Phe Thr Pro Pro Ser Ile Leu Arg Ser Gly Phe Arg Thr Thr Pro 225 230 235 240 Leu Ala Ser Pro Ser Leu Ser Pro Gly Arg Ser Leu Thr Pro Pro Phe 245 250 255 Arg Val Lys Glu Thr Arg Ile Ser Phe Met Glu Glu Gly Met Asn Thr 260 265 270 His Trp Thr Asp Arg Ala Thr Asp Asp Arg Asn Thr Lys Ala Phe Val 275 280 285 Ser Thr Ser Phe His Lys Cys Gly Leu Pro Ala Glu Thr Glu Trp Met 290 295 300 Lys Thr Ser Asp Lys Asn Thr Tyr Phe Pro Leu Asp Val Pro Ala Lys 305 310 315 320 Gly Pro Gln Lys Val Val Ala Glu Ser Leu Ala Thr His Ser Gly Arg 325 330 335 Leu Glu Lys Leu Asp Val Ser Lys Glu Asp Ser Thr Ala Ser Thr Arg 340 345 350 Ser Asp Gln Thr Ser Leu Glu Tyr His Asp Ala Pro Ser Pro Glu Asp 355 360 365 Leu Glu Gly Ala Val Phe Val Ser Pro Lys Pro Ala Ser Ser Ser Thr 370 375 380 Glu Leu Thr Thr Asn Ser Thr Leu Gln Thr Glu Arg Asp Asn Asp Lys 385 390 395 400 Asp Ala Phe Lys Ser Glu Gly Thr Pro Ser Pro Val Lys Lys Gln Ile 4 05 410 415 Gly Thr Gly Asp Ala Ala Val Glu Ala Phe Ser Glu Leu Ser Arg Leu 420 425 430 Asp Pro Val Glu Arg Ala Glu Ala Ser Phe Gly Val Ser Ser Val Cys 435 440 445 Glu Gly Glu Thr Ser Thr Ser Asn Ser Lys Thr Ser Val Leu Asp Gly 450 455 460 Ile Val Pro Ile Glu Ser Arg Thr Ser Ile Leu Thr Ala Asp His Lys 465 470 475 480 Glu Ser Val Ala Asn Thr Val Ala Asp Val Glu Ser Ser Gly Ser Thr 485 490 495 Ser Ser Lys Cys Pro Val Thr Ser Glu Arg Ser Leu Gly Gln Lys Leu 500 505 510 Thr Leu Asn Leu Lys Glu Asp Glu Ile Glu Ala His Val Pro Lys Glu 515 520 525 Asn Val Gly Leu Pro Glu Glu Ser Pro Arg Ile Ser Ala Ala Pro Ser 530 535 540 Asp Thr His Glu Ile His Leu Ile Gly Cys Glu Asn Leu Glu Val Gln 545 550 555 560 Asn Ser Glu Glu Glu Ala Lys Asn Leu Ser Phe Asp Glu Leu Tyr Pro 565 570 575 Leu Gly Ala Glu Lys Leu Glu Tyr Asn Leu Ser Thr Ile Glu Gln Gln 580 585 590 Phe Cys Asp Leu Pro Asp Asp Lys Asp Ser Ala Glu Cys Asp Ala Ala 595 600 605 Glu Val Asp Gly Glu Leu Phe Val Ala Gln Ser Asn Phe Thr Leu Ile 610 615 620 Leu Glu Gly Glu Glu Gly Glu Ala Glu Ala Ser Asp Ser Ala Ala Pro 625 630 635 640 Asn Met Leu Pro Lys Ser Thr Lys Glu Lys Pro Val Cys Tyr Arg Glu 645 650 655 Pro His Asn Gln Glu Arg Val Thr Asp Leu Pro Ser Ala Val Thr Ala 660 665 670 Asp Gln Glu Ser His Lys Val Glu Thr Leu Pro Tyr Val Pro Glu Pro 675 680 685 Val Lys Val Ala Ile Ala Glu Asn Leu Leu Asp Val Ile Lys Asp Thr 690 695 700 Arg Ser Lys Glu Ala Thr Pro Val Ala Ala Gly Glu Ala Gly Asp Glu 705 710 710 720 Asp Gly Ala Val Ile Val Ser Lys Ala Ala His Ser Ser Arg Leu Thr 725 730 735 Asn Ser Thr Pro Lys Thr Val Lys Glu Pro Arg Ala Glu Thr Val Asn 740 745 750 Thr Ser Gln Ser Asp Asp Met Val Ser Ser Arg Thr Leu Thr Arg Arg 755 760 765 Gln His Ala Leu Ser Leu Asn Val Thr Ser Glu Gln Glu Pro Ser Ala 770 775 780 val Ala Thr Pro Lys Lys Arg Thr Arg Lys Ile Lys Glu Thr Pro Glu 785 790 795 800 Ser Ser Glu Arg Thr Cys Ser Asp Leu Lys Val Ala Pro Glu Asn Gln 805 810 815 Leu Thr Ala Gln Asn Pro Pro Ala Pro Arg Ar g Arg Lys Lys Lys Asp 820 825 830 Val Ser Gln Gly Thr Leu Pro Ser Ser Gly Ala Val Glu Pro Glu Pro 835 840 845 Glu Pro Gln Gly Thr Pro Gly Arg Leu Arg Leu Arg Thr Gln Pro Pro 850 855 860 Glu Pro Ala Ala Glu Glu Thr Pro Ser Arg Thr Lys Val Arg Leu Ser 865 870 875 880 Ser Val Arg Lys Gly Thr Pro Arg Arg Leu Lys Lys Ser Val Glu Asn 885 890 895 Gly Gln Ser Ile Glu Ile Leu Asp Asp Leu Lys Gly Ser Glu Ala Ala 900 905 910 Ser His Asp Gly Thr Val Thr Glu Leu Arg Asn Ala Asn Leu Glu Asp 915 920 925 Thr Gln Asn Met Glu Tyr Lys Gln Asp Glu His Ser Asp Gln Gln Pro 930 935 940 Pro Leu Lys Arg Lys Arg Val Arg Glu Arg Glu Val Ser Val Ser Ser 945 950 955 960 Val Thr Glu Glu Pro Lys Leu Asp Ser Ser Gln Leu Pro Leu Gln Thr 965 970 975 Gly Leu Asp Val Pro Ala Thr Pro Arg Lys Arg Gly Arg Pro Arg Lys 980 985 990 Val Val Pro Leu Glu Ala Asp Gly Gly Thr Thr Gly Lys Glu Gln Thr 995 1000 1005 Ser Pro Gln Lys Lys Asp Val Pro Val Val Arg Arg Ser Thr Arg Asn 1010 1015 1020 Thr Pro Ala Arg Asn Val Ser Thr Leu Xaa Lys Ser Val Leu Val Pro 1025 1030 1035 1040 Asn Lys Glu Ala Ala Leu Val Val Thr Ser Lys Arg Arg Pro Thr Lys 1045 1050 1055 Lys Ser Ala Glu Glu Ser Ser Lys Asp Pro Ser Ala Ala Val Ser Asp 1060 1065 1070 Trp Ala Gly Gly Ala Ala His Thr Glu Ser Ala Asp Arg Arg Asp Gly 1075 1080 1085 Leu Leu Ala Ala Ala Ala Leu Thr Pro Ser Ala Gln Gly Thr Arg Thr 1090 1095 1100 Arg Ser Arg Arg Thr Met Leu Leu Thr Asp Ile Ser Glu Pro Lys Thr 1105 1110 1115 1120 Glu Pro Leu Phe Pro Pro Pro Ser Val Lys Val Pro Lys Lys Lys Ser 1125 1130 1135 Lys Ala Glu Asn Met Glu Ala Ala Ala Gln Leu Lys Glu Leu Val Ser 1140 1145 1150 Asp Leu Ser Ser Gln Phe Val Val Ser Pro Pro Ala Leu Arg Thr Arg 1155 1160 1165 Gln Lys Ser Ile Ser Asn Thr Ser Lys Leu Leu Gly Glu Leu Glu Ser 1170 1175 1180 Asp Pro Lys Pro Leu Glu Ile Ile Glu Gln Lys Pro Lys Arg Ser Arg 1185 1190 1195 1200 Thr Val Lys Thr Arg Ala Ser Arg Asn Thr Gly Lys Gly Ser Ser Trp 1205 1210 1215 Ser Pro Pro Pro Val Glu Ile Lys Leu Val Ser Pro Leu A la Ser Pro 1220 1225 1230 Val Asp Glu Ile Lys Thr Gly Lys Pro Arg Lys Thr Ala Glu Ile Ala 1235 1240 1245 Gly Lys Thr Leu Gly Arg Gly Arg Lys Lys Pro Ser Ser Phe Pro Lys 1250 1255 1260 Gln Ile Leu Arg Arg Lys Met Leu 1265 1270 <210> 3 <211> 24 <212> DNA <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Artificially Synthesized Primer Sequence <400> 3 cacccgtgaa gaaacaaata ggca 24 <210> 4 <211> 24 <212> DNA <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Artificially Synthesized Primer Sequence <400> 4 cctttggtac atgagcttct attt 24 <210> 5 <211> 4883 <212> DNA <213> Homo sapiens <220> <221> CDS <222> (1) .. (4590) <400> 5 gag aag ttg tgg aaa cga gat gaa gga ggc aca gga aaa tat cct cct 48 Glu Lys Leu Trp Lys Arg Asp Glu Gly Gly Thr Gly Lys Tyr Pro Pro 1 5 10 15 gct agt ctg cat gca gta ctt gat atg tac cta tta gac ggc gtt act 96 Ala Ser Leu His Ala Val Leu Asp Met Tyr Leu Leu Asp Gly Val Thr 20 25 30 gaa gca gcc aaa cac tct att acc att tat ttg cta ctt gat att atg 144 Glu Ala Ala Lys His Ser Ile Thr Ile Tyr Leu Leu Leu Asp Ile Met 35 40 45 tat tcc ttt ccc aac aaa aca gac act ccc att gaa tct ttc cca act 192 Tyr Ser Phe Pro Asn Lys Thr Asp Thr Pro Ile Glu Ser Phe Pro Thr 50 55 60 gta ttt gcc att tct tgg ggc caa gtt aaa ctt att cag ggg ttt tgg 240 Val Phe Ala Ile Ser Trp Gly Gln Val Lys Leu Ile Gln Gly Phe Trp 65 70 75 80 ttg ata gat cat aat gac tat gag agt ggt ttg gat ctt ttg ttt cat 288 Leu Ile Asp His Asn Asp Tyr Glu Ser Gly Leu Asp Leu Leu Phe His 85 90 95 cca gct act gca aaa cct ttg tca tgg caa cat tca aag att att cag 336 Pro Ala Thr Ala Lys Pro Leu Ser Trp Gln His Ser Lys Ile Ile Gln 100 10 5 110 gca ttc atg agt cag ggc gag cac aga caa gcc ctc aga tat att cag 384 Ala Phe Met Ser Gln Gly Glu His Arg Gln Ala Leu Arg Tyr Ile Gln 115 120 125 aca atg aag cca aca gtg tcc agt ggt aac gat gtt atc ctt cac ctc 432 Thr Met Lys Pro Thr Val Ser Ser Gly Asn Asp Val Ile Leu His Leu 130 135 140 act gtt ttg ctt ttt aat agg tgt atg gtt gaa gcc tgg aat ttt ttg 480 Thr Val Leu Leu Phe Asn Arg Cys Met Val Glu Ala Trp Asn Phe Leu 145 150 155 160 cgg caa cat tgc aat agg ttg aat ata gag gag tta ctg aag cac atg 528 Arg Gln His Cys Asn Arg Leu Asn Ile Glu Glu Leu Leu Lys His Met 165 170 175c tat gaagt tgt cag gaa atg ggc ttg atg gaa gat tta ctg aag tta 576 Tyr Glu Val Cys Gln Glu Met Gly Leu Met Glu Asp Leu Leu Lys Leu 180 185 190 cca ttt aca gac act gag cag gaa tgt tta gtg aaa ttt 624 Pro Phe Thr Asp Thr Glu Gln Glu Cys Leu Val Lys Phe Leu Gln Ser 195 200 205 agt gcc agc gtt cag aat cat gaa ttc ctt tta gtg cac cat ttg cag 672 Ser Ala Ser Val Gln Asn His Glu Phe Leu Leu Val His His Leu Gln 210 215 220 cgt gcc aat tat gtg cct gcc ttg aag ctg aac caa act ctg aag att 720 Arg Ala Asn Tyr Val Pro Ala Leu Lys Leu Asn Gln Thr Leu Lys Ile 225 230 235 240 aat gtt atg aat gat cgt gat cct cgt ttg cgg gag aga tca ctg gct 768 Asn Val Met Asn Asp Arg Asp Pro Arg Leu Arg Glu Arg Ser Leu Ala 245 250 255 cga aat tct ata tta gac cag tat gga aaa atc ctt cct aga gtc cat 816 Arg Asn Ser Ile Leu Asp Gln Tyr Gly Lys Ile Leu Pro Arg Val His 260 265 270 cga aaa tta gcc att gaa cga gct aag cct tat cat ctg tca aca tca 864 Arg Lys Leu Ala Ile Glu Arg Ala Lys Pro Tyr His Leu Ser Thr Ser 275 280 285 tca gtt ttt cga tta gtt tct aga ccc aaa cca tta tca gca gtt cca 912 Ser Val Phe Arg Leu Val Ser Arg Pro Lys Pro Leu Ser Ala Val Pro 290 295 300 aag caa gtt gta aca gga act gtg ttg aca aga tct gtt ttc atc aac 960 Lys Gln Val Val Thr Gly Thr Val Leu Thr Arg Ser Val Phe Ile Asn 305 310 315 320 aat gtg tta tct aaa att gga gaa gtt tgg gca agc aaa gaa cct ata 1008 Asn Val Leu Ser Lys Ile Gly Glu Val Trp Ala Ser Lys Glu Pro Ile 325 330 335 aat agc acc aca cct ttc aat agt tct aaa ata gaa gaa cca tct cct 1056 Asn Ser Thr Thr Thr Pro Phe Asn Ser Ser Lys Ile Glu Glu Pro Ser Pro 340 345 350 350 ata gtg tat tcg ctc cca gct cca gag ctg cct gag gca ttt ttt gga 1104 Ile Val Tyr Ser Leu Pro Ala Pro Glu Leu Pro Glu Ala Phe Phe Gly 355 360 365 aca cca att tca aaa gca tca caa aaa att tct aga ctg cta gatg 1152 Thr Pro Ile Ser Lys Ala Ser Gln Lys Ile Ser Arg Leu Leu Asp Leu 370 375 380 gtt gtt cag cct gtc ccc cgg cct tct cag tgt tcg gag ttt att cag 1200 Val Val Gln Pro Val Pro Arg Pro Ser Gln Cys Ser Glu Phe Ile Gln 385 390 395 400 400 caa agc tcc atg aaa tct cct ttg tac cta gta tcc cgt tca ctg ccc 1248 Gln Ser Ser Met Lys Ser Pro Leu Tyr Leu Val Ser Arg Ser Leu Pro 405 410 415 tca agt tcg caa tta aaa gga tcg cct cag gcc atc tcc agg gct tca 1296 Ser Ser Ser Gln Leu Lys Gly Ser Pro Gln Ala Ile Ser Arg Ala Ser 420 425 430 gaa tta cat ttg ctt gaa act cct ctt gta gtt aag aaa gct aaa agt 1344 Gl u Leu His Leu Leu Glu Thr Pro Leu Val Val Lys Lys Ala Lys Ser 435 440 445 ttg gcc atg tca gtt act act tct gga ttt tct gag ttc act cct cag 1392 Leu Ala Met Ser Val Thr Thr Ser Gly Phe Ser Glu Phe Thr Pro Gln 450 455 460 tcc atc ctg agg tct act cct cga tca aca cct tta gca tct ccc tct 1440 Ser Ile Leu Arg Ser Thr Pro Arg Ser Thr Pro Leu Ala Ser Pro Ser 465 470 475 475 480 cca tca cct gga agg tct cct caa cga ctt aaa gaa act aga att tca 1488 Pro Ser Pro Gly Arg Ser Pro Gln Arg Leu Lys Glu Thr Arg Ile Ser 485 490 495 ttt gtg gaa gaa gat gtc cac cca aaa tgg att cct ggg gct gca gat 1536 Phe Val Glu Glu Asp Val His Pro Lys Trp Ile Pro Gly Ala Ala Asp 500 505 510 gat agc aaa tta gaa gta ttt act aca cct aaa aaa tgt gca gtt cca 1584 Asp Ser Lys Leu Glu Val Phe Thr Thr Pro Lys Lys Cys Ala Val Pro 515 520 525 gtg gaa act gaa tgg ccg aag agc aaa gat agg acc aca tct ttt ttc 1632 Val Glu Thr Glu Trp Pro Lys Ser Lys Asp Arg Thr Thr Ser Phe Phe 530 535 540 ctg aac agc cct gaa aag gag cat caa gaa atg g at gag ggg tca caa 1680 Leu Asn Ser Pro Glu Lys Glu His Gln Glu Met Asp Glu Gly Ser Gln 545 550 555 560 agt tta gag aaa ctg gat gtg agc aaa gga aac agc agt gtt tca atc 1728 Ser Leu Glu Lys Leu Asp Val Ser Lys Gly Asn Ser Ser Val Ser Ile 565 570 575 aca tcc gat gag act acc tta gag tat cag gat gca ccg tca ccg gaa 1776 Thr Ser Asp Glu Thr Thr Leu Glu Tyr Gln Asp Ala Pro Ser Pro Glu 580 585 585 590 gac ctt gaa gag act gtt ttc acg gcc tct aag ccc aaa agc tct tcc 1824 Asp Leu Glu Glu Thr Val Phe Thr Ala Ser Lys Pro Lys Ser Ser Ser 595 600 605 act gca cta act act aat gta act gaa caa act gaa aag gat gga gat 1872 Thr Ala Leu Thr Thr Asn Val Thr Glu Gln Thr Glu Lys Asp Gly Asp 610 615 620 aaa gat gta ttt gca tca gaa gta act cct tca gac cta cag aaa caa 1920 Lys Asp Val Phe Ala Ser Glu Val Thr Pro Ser Asp Leu Gln Lys Gln 625 630 635 640 atg ggc aat tta gaa gat gca gaa aca aag gat ctc tta gtt gca gca 1968 Met Gly Asn Leu Glu Asp Ala Glu Thr Lys Asp Leu Leu Val Ala Ala 645 650 655 gag gca ttt tca gaa ttg aat cac tta agc ccg gtt caa gga act gaa 2016 Glu Ala Phe Ser Glu Leu Asn His Leu Ser Pro Val Gln Gly Thr Glu 660 665 670 gct tct ctt tgt gca cca tca gtc tat gaa ggg aaa atc ttc acc cag 2064 Ser Leu Cys Ala Pro Ser Val Tyr Glu Gly Lys Ile Phe Thr Gln 675 680 685 aag tcc aag gta cca gtg ttg gac gaa gga tta aca tct gtt gaa acc 2112 Lys Ser Lys Val Pro Val Leu Asp Glu Gly Leu Thr Ser Val Glu Thr 690 695 700 tac acc cct gca att aga gca aat gac aat aaa tct atg gct gat gtc 2160 Tyr Thr Pro Ala Ile Arg Ala Asn Asp Asn Lys Ser Met Ala Asp Val 705 710 715 715 720 ctt ggt gat ggt gga aac tcc tcg ctc act atc tct gaa ggt cct att 2208 Leu Gly Asp Gly Gly Asn Ser Ser Leu Thr Ile Ser Glu Gly Pro Ile 725 730 735 gtc tct gag cgc agg ctt aac cag gaa gta gcg ctg aac tta aaa gaa 2256 Val Ser Glu Arg Arg Asn Gln Glu Val Ala Leu Asn Leu Lys Glu 740 745 750 gat cat gaa gta gaa gtt ggt gta cta aaa gaa agt gtt gac tta cca 2304 Asp His Glu Val Glu Val Gly Val Leu Lys Glu Ser Val Asp Leu Pro 75 5 760 765 gaa gaa aag ctt cca att tct gac agc cct cct gat act caa gaa att 2352 Glu Glu Lys Leu Pro Ile Ser Asp Ser Pro Pro Asp Thr Gln Glu Ile 770 775 780 cat gtg att gaa caa gaa aag ctt gaga gct gat tca gga gaa gag 2400 His Val Ile Glu Gln Glu Lys Leu Glu Ala Gln Asp Ser Gly Glu Glu 785 790 795 800 gct agg aat ctt tca ttt aat gag tta tat ccc tct gga aca ctt aag 2448 Ala Arg Asn Leu Ser Phe Asn Glu Leu Tyr Pro Ser Gly Thr Leu Lys 805 810 815 ctt cag tac aat ttt gat act att gac caa cag ttt tgt gac tta gct 2496 Leu Gln Tyr Asn Phe Asp Thr Ile Asp Gln Gln Phe Cys Asp Leu Ala 820 825 830 gat aac aaa gac act gct gaa tgt gac att gct gaa gta gat ggg gaa 2544 Asp Asn Lys Asp Thr Ala Glu Cys Asp Ile Ala Glu Val Asp Gly Glu 835 840 845 ctt ttt gtg gct caa agc aac ttt acc ttg ata gtgga g 2592 Leu Phe Val Ala Gln Ser Asn Phe Thr Leu Ile Leu Glu Gly Glu Glu 850 855 860 gga gaa gtt gag cca ggt gat ttt gca tca tct gat gtg tta cct aaa 2640 Gly Glu Val Glu Pro Gly Asp Phe Ala Ser S er Asp Val Leu Pro Lys 865 870 875 880 gca gct aac aca gca act gaa gaa aaa ctt gta tgc agt ggg gaa aat 2688 Ala Ala Asn Thr Ala Thr Glu Glu Lys Leu Val Cys Ser Gly Glu Asn 885 890 890 895 gat aat cat gga caa att gca aat ttg cca tct gcc gta act agt gac 2736 Asp Asn His Gly Gln Ile Ala Asn Leu Pro Ser Ala Val Thr Ser Asp 900 905 910 caa aag tcc caa aaa gta gac act tta cca tat gtg cct gaa cct att 2784 Gln Lys Ser Gln Lys Val Asp Thr Leu Pro Tyr Val Pro Glu Pro Ile 915 920 925 aaa gta gca att gca gaa aat tta cta gat gta att aaa gac aca aga 2832 Lys Val Ala Ile Ala Glu Asn Leu Leu Asp Val Ile Lys Asp Thr Arg 930 935 940 agt aaa gaa att act tca gat aca atg gaa cag tcc att cat gaa aca 2880 Ser Lys Glu Ile Thr Ser Asp Thr Met Glu Gln Ser Ile His Glu Thr 945 950 955 960 ata cct tta gtg agc caa aac ata atg tgt ccc act aaa ttg gtc aaa 2928 Ile Pro Leu Val Ser Gln Asn Ile Met Cys Pro Thr Lys Leu Val Lys 965 970 975 tct gca ttt aag act gct cag gaa aca agc aca atg act atg aat gtc 2976 Ser Ala Phe Lys Thr Ala Gln Glu Thr Ser Thr Met Thr Met Asn Val 980 985 990 agc cag gtt gat gac gtg gtt tcc tcc aaa act cgt acg aga ggt caa 3024 Ser Gln Val Asp Asp Val Val Ser Ser Lys Thr Arg Thr Arg Gly Gln 995 1000 1005 cgt atc caa aac gtg aat gtc aaa tca gca caa cag gaa gca tca gca 3072 Arg Ile Gln Asn Val Asn Val Lys Ser Ala Gln Gln Glu Ala Ser Ala 1010 1015 1020 gat gtt gct act cct aag atg cca ggg cag tca gt agg aag aaa act 3120 Asp Val Ala Thr Pro Lys Met Pro Gly Gln Ser Val Arg Lys Lys Thr 1025 1030 1035 1040 agg aag gca aaa gaa att tct gaa gct tct gaa aac atc tat tct gat 3168 Arg Lys Ala Lys Glu Ile Ser Glu Ala Ser Glu Asn Ile Tyr Ser Asp 1045 1050 1055 gtc aga gga cta ttt cag aac cag caa ata cct caa aat tct gtt acg 3216 Val Arg Gly Leu Phe Gln Asn Gln Gln Ile Pro Gln Asn Ser Val Thr 1060 1065 1070 cct agg aga gga agg aga aag aaa gaa gtt aat cag gac ata cta gaa 3264 Pro Arg Arg Gly Arg Arg Lys Lys Glu Val Asn Gln Asp Ile Leu Glu 1075 1080 1085 aac acc agt tct gtg gaa caa gaa tta cag atc act aca ggt agg gaa 3312 Asn Thr Ser Ser Val Glu Gln Glu Leu Gln Ile Thr Thr Gly Arg Glu 1090 1095 1100 tca aaa aga tta aaa aaa tca tct cag ctg ttg gaa cca gca gtt gaa gaa 3360 Ser Lys Arg Leu Lys Ser Ser Gln Leu Leu Glu Pro Ala Val Glu Glu 1105 1110 1115 1120 act act aaa aaa gaa gtt aag gtt tca tct gtt aca aaa agg act cct 3408 Thr Thr Lys Lys Glu Val Lys Val Ser Ser Val Thr Lys Arg Thr Pro 1125 1130 1135 aga aga att aaa aga tct gta gaa aat cag gaa agt gtt gaa att ata 3456 Arg Arg Ile Lys Arg Ser Val Glu Asn Gln Glu Ser Val Glu Ile Ile 1140 1145 1150 aat gat cta aaa gtt agt acg gta aca agt cct agc aga atc aga 3504 Asn Asp Leu Lys Val Ser Thr Val Thr Thr Ser Pro Ser Arg Met Ile Arg 1155 1160 1165 aaa ttg aga agt act aat tta gat gct tct gaa aat aca gga aat aag 3552 Lys Leu Arg Ser Thr Asn Leu Asp Ala Ser Glu Asn Thr Gly Asn Lys 1170 1175 1180 caa gat gat aaa tcc agt gac aag cag ctg cgt att aaa cat gtt aga 3600 Gln Asp Asp Lys Ser Ser Asp Lys Gln Leu Arg Ile Lys His Val Arg 1185 1190 1195 1200 agg gtc aga ggg aga gaa gtt agt cca tca gat gtg aga gaa gac tcc 3648 Arg Val Arg Gly Arg Glu Val Ser Pro Ser Asp Val Arg Glu Asp Ser 1205 1210 1215 aac ctt gag tca tct cag ttg act gtt caa gca gaa ttt gat atg tct 3696 Asn Leu Glu Ser Ser Gln Leu Thr Val Gln Ala Glu Phe Asp Met Ser 1220 1225 1230 gcc ata cct aga aaa cgt ggt aga cca aga aaa atc aat cca tct gaa 3744 Ala Ile Pro Arg Lys Arg Gly Arg Pro Arg Lys Ile Asn Pro Ser Glu 1235 1240 1245 gat gta gga tct aag gct gtt aag gaa gag aga agc ccc aag aag aaa 3792 Asp Val Gly Ser Lys Ala Val Lys Glu Glu Arg Ser Pro Lys Lys Lys 1250 1255 1260 gaa gct ccc agc att aga agg aga tct aca aga aat acc cca gct aaa 3840 Glu Ala Pro Ser Ile Arg Arg Arg Ser Thr Arg Asn Thr Pro Ala Lys 1265 1270 1275 1280 agt gaa aat gtt gat gtt gga aaa cca gct tta gga aaa tcc att tta 3888 Ser Glu Asn Val Asp Val Gly Lys Pro Ala Leu Gly Lys Ser Ile Leu 1285 1290 1295 gtg cca aac gag gaa ctt tcg atg gtg atg ag agc tct aag aaa aaa ctt 3936 Val Pro Asn Glu Glu Leu Ser Met Val Met Ser Ser Lys Lys Lys Leu 1300 1305 1310 aca aaa aag act gaa agt caa agc caa aaa cgt tca ttg cac tca gta 3984 Thr Lys Lys Thr Glu Ser Gln Ser Gln Lys Arg Ser Leu His Ser Val 1315 1320 1325 tca gaa gaa cgc aca gat gaa atg aca cat aaa gaa aca aat gag cag 4032 Ser Glu Glu Arg Thr Asp Glu Met Thr His Lys Glu Thr Asn Glu Gln 1330 1335 1340 gaa gaa aga ttg ctc gcc aca gct tcctt ccc tcc cgc agc 4080 Glu Glu Arg Leu Leu Ala Thr Ala Ser Phe Thr Lys Ser Ser Arg Ser 1345 1350 1355 1360 agc agg act cgg tct agc aag gcc atc ttg ttg ccg gac ctt tct gaa 4128 Ser Arg Thr Arg Ser Ser Lys Ala Ile Leu Leu Pro Asp Leu Ser Glu 1365 1370 1375 cca aac aat gag cct tta ttt tct cca gcg tca gaa gtt cca agg aaa 4176 Pro Asn Asn Glu Pro Leu Phe Ser Pro Ala Ser Glu Val Pro Arg Lys 1380 1385 1390 gca aaa gct aaa aaa ata gag gtt cct gca cag ctg aaa gaa tta gtt 4224 Ala Lys Ala Lys Lys Ile Glu Val Pro Ala Gln Leu Lys Glu Leu Val 1395 1400 1405 tcg gat tta tct tct cag ttt gtc atc t ca cct cct gct tta agg agc 4272 Ser Asp Leu Ser Ser Gln Phe Val Ile Ser Pro Pro Ala Leu Arg Ser 1410 1415 1420 aga caa aaa aac aca tcc aat aag aac aag ctt gaa gat gaa ctg aaa 4320 Arg Gln Lys Asn Thr Ser Asn Lys Asn Lys Leu Glu Asp Glu Leu Lys 1425 1430 1435 1440 gat gat gca caa tca gta gaa act ctg gga aag cca aaa gcg aaa cga 4368 Asp Asp Ala Gln Ser Val Glu Thr Leu Gly Lys Pro Lys Ala Lys Arg 1445 1455 atc agg acg tca aaa aca aaa caa gca agc aaa aac aca gaa aaa gaa 4416 Ile Arg Thr Ser Lys Thr Lys Gln Ala Ser Lys Asn Thr Glu Lys Glu 1460 1465 1470 agt gct tgg tca ctt cct ccc ata gaa att cgg ccgt ccc ttg 4464 Ser Ala Trp Ser Leu Pro Pro Ile Glu Ile Arg Leu Ile Ser Pro Leu 1475 1480 1485 gct agc cca gct gac gga gtc aag agc aaa cca aga aaa act aca gaa 4512 Ala Ser Pro Ala Asp Gly Val Lys Ser Lys Pro Arg Lys Thr Thr Glu 1490 1495 1500 gtg aca gga aca ggt ctt gga agg aac aga aag aaa ctg tct tcc tat 4560 Val Thr Gly Thr Gly Leu Gly Arg Asn Arg Lys Lys Leu Ser Ser Tyr 1505 1510 1515 1520 cca aag caa att tta cgc aga aaa atg ctg taatttcttg ggaagatttt 4610 Pro Lys Gln Ile Leu Arg Arg Lys Met Leu 1525 1530 aatgtacacc tatttgtaaa gtcatcagaa tagtgtggat tattaaatat ctagtttgga 4670 agaaaataat ttatataaat tattgtaaat ttttatgtaa acagaaggtc ttcaataagt 4730 aaagtaactc catatggagt gattgtttca gtccaggcaa tttttctatt ttatattaag 4790 acttcataca tttatatatg taaatatggc ttattaatgg aatgttaaat aaaatgtata 4850 cttctcaaaa aaaaaaaaaa aaaaaaaaaa aaa 4883 <210> 6 <211> 1530 <212> PRT <213> Homo sapiens <400> 6 Glu Lys Leu Trp Lys Arg Asp Glu Gly Gly Thr Gly Lys Tyr Pro Pro 1 5 10 15 Ala Ser Leu His Ala Val Leu Asp Met Tyr Leu Leu Asp Gly Val Thr 20 25 30 Glu Ala Ala Lys His Ser Ile Thr Ile Tyr Leu Leu Leu Asp Ile Met 35 40 45 Tyr Ser Phe Pro Asn Lys Thr Asp Thr Pro Ile Glu Ser Phe Pro Thr 50 55 60 Val Phe Ala Ile Ser Trp Gly Gln Val Lys Leu Ile Gln Gly Phe Trp 65 70 75 80 Leu Ile Asp His Asn Asp Tyr Glu Ser Gly Leu Asp Leu Leu Phe His 85 90 95 Pro Ala Thr Ala Lys Pro Leu Ser Trp Gln His Ser Lys Ile Ile Gln 100 105 110 Ala Phe Met Ser Gln Gly Glu His Arg Gln Ala Leu Arg Tyr Ile Gln 115 120 125 Thr Met Lys Pro Thr Val Ser Ser Gly Asn Asp Val Ile Leu His Leu 130 135 140 Thr Val Leu Leu Phe Asn Arg Cys Met Val Glu Ala Trp Asn Phe Leu 145 150 155 160 Arg Gln His Cys Asn Arg Leu Asn Ile Glu Glu Leu Leu Lys His Met 165 170 175 Tyr Glu Val Cys Gln Glu Met Gly Leu Met Glu Asp Leu Leu Lys Leu 180 185 190 Pro Phe Thr Asp Thr Glu Gln Glu Cys Leu Val Lys Phe Leu Gln Ser 195 200 205 Se r Ala Ser Val Gln Asn His Glu Phe Leu Leu Val His His Leu Gln 210 215 220 Arg Ala Asn Tyr Val Pro Ala Leu Lys Leu Asn Gln Thr Leu Lys Ile 225 230 235 240 Asn Val Met Asn Asp Arg Asp Pro Arg Leu Arg Glu Arg Ser Leu Ala 245 250 255 Arg Asn Ser Ile Leu Asp Gln Tyr Gly Lys Ile Leu Pro Arg Val His 260 265 270 Arg Lys Leu Ala Ile Glu Arg Ala Lys Pro Tyr His Leu Ser Thr Ser 275 280 285 Ser Val Phe Arg Leu Val Ser Arg Pro Lys Pro Leu Ser Ala Val Pro 290 295 300 Lys Gln Val Val Thr Gly Thr Val Leu Thr Arg Ser Val Phe Ile Asn 305 310 315 320 Asn Val Leu Ser Lys Ile Gly Glu Val Trp Ala Ser Lys Glu Pro Ile 325 330 335 Asn Ser Thr Thr Pro Phe Asn Ser Ser Lys Ile Glu Glu Pro Ser Pro 340 345 345 350 Ile Val Tyr Ser Leu Pro Ala Pro Glu Leu Pro Glu Ala Phe Phe Gly 355 360 365 Thr Pro Ile Ser Lys Ala Ser Gln Lys Ile Ser Arg Leu Leu Asp Leu 370 375 380 Val Val Gln Pro Val Pro Arg Pro Ser Gln Cys Ser Glu Phe Ile Gln 385 390 395 400 Gln Ser Ser Met Lys Ser Pro Leu Tyr Leu Val Ser Arg Ser Leu Pro 40 5 410 415 Ser Ser Ser Gln Leu Lys Gly Ser Pro Gln Ala Ile Ser Arg Ala Ser 420 425 430 Glu Leu His Leu Leu Glu Thr Pro Leu Val Val Lys Lys Ala Lys Ser 435 440 445 Leu Ala Met Ser Val Thr Thr Ser Gly Phe Ser Glu Phe Thr Pro Gln 450 455 460 Ser Ile Leu Arg Ser Thr Pro Arg Ser Thr Pro Leu Ala Ser Pro Ser 465 470 475 480 480 Pro Ser Pro Gly Arg Ser Pro Gln Arg Leu Lys Glu Thr Arg Ile Ser 485 490 495 Phe Val Glu Glu Asp Val His Pro Lys Trp Ile Pro Gly Ala Ala Asp 500 505 510 Asp Ser Lys Leu Glu Val Phe Thr Thr Pro Lys Lys Cys Ala Val Pro 515 520 525 Val Glu Thr Glu Trp Pro Lys Ser Lys Asp Arg Thr Thr Ser Phe Phe 530 535 540 Leu Asn Ser Pro Glu Lys Glu His Gln Glu Met Asp Glu Gly Ser Gln 545 550 555 560 Ser Leu Glu Lys Leu Asp Val Ser Lys Gly Asn Ser Ser Val Ser Ile 565 570 575 575 Thr Ser Asp Glu Thr Thr Leu Glu Tyr Gln Asp Ala Pro Ser Pro Glu 580 585 590 Asp Leu Glu Glu Thr Val Phe Thr Ala Ser Lys Pro Lys Ser Ser Ser 595 600 605 Thr Ala Leu Thr Thr Asn Val Thr Glu Gln Thr Glu Lys Asp G ly Asp 610 615 620 Lys Asp Val Phe Ala Ser Glu Val Thr Pro Ser Asp Leu Gln Lys Gln 625 630 635 640 Met Gly Asn Leu Glu Asp Ala Glu Thr Lys Asp Leu Leu Val Ala Ala 645 650 655 Glu Ala Phe Ser Glu Leu Asn His Leu Ser Pro Val Gln Gly Thr Glu 660 665 670 Ala Ser Leu Cys Ala Pro Ser Val Tyr Glu Gly Lys Ile Phe Thr Gln 675 680 685 Lys Ser Lys Val Pro Val Leu Asp Glu Gly Leu Thr Ser Val Glu Thr 690 695 700 Tyr Thr Pro Ala Ile Arg Ala Asn Asp Asn Lys Ser Met Ala Asp Val 705 710 715 720 Leu Gly Asp Gly Gly Asn Ser Ser Leu Thr Ile Ser Glu Gly Pro Ile 725 730 735 Val Ser Glu Arg Arg Leu Asn Gln Glu Val Ala Leu Asn Leu Lys Glu 740 745 750 Asp His Glu Val Glu Val Gly Val Leu Lys Glu Ser Val Asp Leu Pro 755 760 765 Glu Glu Lys Leu Pro Ile Ser Asp Ser Pro Pro Asp Thr Gln Glu Ile 770 775 780 His Val Ile Glu Gln Glu Lys Leu Glu Ala Gln Asp Ser Gly Glu Glu 785 790 795 800 Ala Arg Asn Leu Ser Phe Asn Glu Leu Tyr Pro Ser Gly Thr Leu Lys 805 810 815 Leu Gln Tyr Asn Phe Asp Thr Ile Asp Gln Gln Phe Cys Asp Leu Ala 820 825 830 Asp Asn Lys Asp Thr Ala Glu Cys Asp Ile Ala Glu Val Asp Gly Glu 835 840 845 Leu Phe Val Ala Gln Ser Asn Phe Thr Leu Ile Leu Glu Gly Glu Glu 850 855 860 Gly Glu Val Glu Pro Gly Asp Phe Ala Ser Ser Asp Val Leu Pro Lys 865 870 875 880 Ala Ala Asn Thr Ala Thr Glu Glu Lys Leu Val Cys Ser Gly Glu Asn 885 890 895 Asp Asn His Gly Gln Ile Ala Asn Leu Pro Ser Ala Val Thr Ser Asp 900 905 910 Gln Lys Ser Gln Lys Val Asp Thr Leu Pro Tyr Val Pro Glu Pro Ile 915 920 925 Lys Val Ala Ile Ala Glu Asn Leu Leu Asp Val Ile Lys Asp Thr Arg 930 935 940 Ser Lys Glu Ile Thr Ser Asp Thr Met Glu Gln Ser Ile His Glu Thr 945 950 955 960 Ile Pro Leu Val Ser Gln Asn Ile Met Cys Pro Thr Lys Leu Val Lys 965 970 975 Ser Ala Phe Lys Thr Ala Gln Glu Thr Ser Thr Met Thr Met Asn Val 980 985 990 Ser Gln Val Asp Asp Val Val Ser Ser Lys Thr Arg Thr Arg Gly Gln 995 1000 1005 Arg Ile Gln Asn Val Asn Val Lys Ser Ala Gln Gln Glu Ala Ser Ala 1010 1015 1020 Asp Val Ala Thr Pro Lys Met Pr o Gly Gln Ser Val Arg Lys Lys Thr 1025 1030 1035 1040 Arg Lys Ala Lys Glu Ile Ser Glu Ala Ser Glu Asn Ile Tyr Ser Asp 1045 1050 1055 Val Arg Gly Leu Phe Gln Asn Gln Gln Ile Pro Gln Asn Ser Val Thr 1060 1065 1070 Pro Arg Arg Gly Arg Arg Lys Lys Glu Val Asn Gln Asp Ile Leu Glu 1075 1080 1085 Asn Thr Ser Ser Val Glu Gln Glu Leu Gln Ile Thr Thr Gly Arg Glu 1090 1095 1100 Ser Lys Arg Leu Lys Ser Ser Gln Leu Leu Glu Pro Ala Val Glu Glu 1105 1110 1115 1120 Thr Thr Lys Lys Glu Val Lys Val Ser Ser Val Thr Lys Arg Thr Pro 1125 1130 1135 Arg Arg Ile Lys Arg Ser Val Glu Asn Gln Glu Ser Val Glu Ile Ile 1140 1145 1150 Asn Asp Leu Lys Val Ser Thr Val Thr Ser Pro Ser Arg Met Ile Arg 1155 1160 1165 Lys Leu Arg Ser Thr Asn Leu Asp Ala Ser Glu Asn Thr Gly Asn Lys 1170 1175 1180 Gln Asp Asp Lys Ser Ser Asp Lys Gln Leu Arg Ile Lys His Val Arg 1185 1190 1195 1200 Arg Val Arg Gly Arg Glu Val Ser Pro Ser Asp Val Arg Glu Asp Ser 1205 1210 1215 Asn Leu Glu Ser Ser Gln Leu Thr Val Gln Ala Glu Phe Asp Met Ser 1220 1225 1230 Ala Ile Pro Arg Lys Arg Gly Arg Pro Arg Lys Ile Asn Pro Ser Glu 1235 1240 1245 Asp Val Gly Ser Lys Ala Val Lys Glu Glu Arg Ser Pro Lys Lys Lys 1250 1255 1260 Glu Ala Pro Ser Ile Arg Arg Arg Ser Thr Arg Asn Thr Pro Ala Lys 1265 1270 1275 1280 Ser Glu Asn Val Asp Val Gly Lys Pro Ala Leu Gly Lys Ser Ile Leu 1285 1290 1295 Val Pro Asn Glu Glu Glu Leu Ser Met Val Met Ser Ser Lys Lys Lys Leu 1300 1305 1310 Thr Lys Lys Thr Glu Ser Gln Ser Gln Lys Arg Ser Leu His Ser Val 1315 1320 1325 Ser Glu Glu Arg Thr Asp Glu Met Thr His Lys Glu Thr Asn Glu Gln 1330 1335 1340 Glu Glu Glu Arg Leu Leu Ala Thr Ala Ser Phe Thr Lys Ser Ser Arg Ser 1345 1350 1355 1360 Ser Arg Thr Arg Ser Ser Lys Ala Ile Leu Leu Pro Asp Leu Ser Glu 1365 1370 1375 Pro Asn Asn Glu Pro Leu Phe Ser Pro Ala Ser Glu Val Pro Arg Lys 1380 1385 1390 Ala Lys Ala Lys Lys Ile Glu Val Pro Ala Gln Leu Lys Glu Leu Val 1395 1400 1405 Ser Asp Leu Ser Ser Gln Phe Val Ile Ser Pro Pro Ala Leu Arg Ser 1410 1415 1420 Arg Gln Lys Asn Thr Ser Asn Lys Asn Lys Leu Glu Asp Glu Leu Lys 1425 1430 1435 1440 Asp Asp Ala Gln Ser Val Glu Thr Leu Gly Lys Pro Lys Ala Lys Arg 1445 1450 1455 Ile Arg Thr Ser Lys Thr Lys Gln Ala Ser Lys Asn Thr Glu Lys Glu 1460 1465 1470 Ser Ala Trp Ser Leu Pro Pro Ile Glu Ile Arg Leu Ile Ser Pro Leu 1475 1480 1485 Ala Ser Pro Ala Asp Gly Val Lys Ser Lys Pro Arg Lys Thr Thr Glu 1490 1495 1500 Val Thr Gly Thr Gly Leu Gly Arg Asn Arg Lys Lys Leu Ser Ser Tyr 1505 1510 1515 1520 Pro Lys Gln Ile Leu Arg Arg Lys Met Leu 1525 1530 <210> 7 <211> 4115 <212> DNA <213> Mus musculus <220> <221> CDS <222> (1) .. (3817) <400> 7 t cag att ctg aag aat aat ctc atg agt gat cgt gac cct cga ttg cgg 49 Gln Ile Leu Lys Asn Asn Leu Met Ser Asp Arg Asp Pro Arg Leu Arg 1 5 10 15 gaa aga tcg gtg act cga aat tct ata tta gac cag tat ggg aaa atc 97 Glu Arg Ser Val Thr Arg Asn Ser Ile Leu Asp Gln Tyr Gly Lys Ile 20 25 30 cta cct aga gtc cag aga aag tta gct gtt gag cga gct aag cct tac 145 Leu Pro Arg Val Gln Arg Lys Leu Ala Val Glu Arg Ala Lys Pro Tyr 35 40 45 cac ctg tcg aca tcc tca gtt ttt cat gaa gtt tct aga ccc aaa ccg 193 His Leu Ser Thr Ser Ser Val Phe His Glu Val Ser Arg Pro Lys Pro 50 55 60 tta tcg gca ttt cca aag aaa gct ata act gga aca gtg tta acc cga 241 Leu Ser Ala Phe Pro Lys Lys Ala Ile Thr Gly Thr Val Leu Thr Arg 65 70 75 80 tct acg ttc atc agc aat gtt tta tct aaa att gga gag gtg tgg gca 289 Ser Thr Phe Ile Ser Asn Val Leu Ser Lys Ile Gly Glu Val Trp Ala 85 90 95 agt cat gag cct aga aat ggc gtc tca ctt ttt aac agt cct aaa aca 337 Ser His Glu Pro Arg Asn Gly Val Ser Leu Phe Asn Ser Pro Lys Thr 100 105 110 gaa cag cca tct cct gta gta cac tct ttc cca cac cca gag ctt cct 385 Glu Gln Pro Ser Pro Val Val His Ser Phe Pro His Pro Glu Leu Pro 115 120 125 gag gcg ttt gtt gga act cca att tca aat aca tcc cag aga att tct 433 Glu Ala Phe Val Gly Thr Pro Ile Ser Asn Thr Ser Gln Arg Ile Ser 130 135 140 aga tta ctg gat ttg gtt gtc cat cct gta ccc cag cct tct cag tgt 481 Arg Leu Leu Asp Leu Val Val His Pro Val Pro Gln Pro Ser Gln Cys 145 150 155 160 ttg gag ttt att caa caa agt ccc aca aga tct cct ttg tgt ctg ctg 529 Leu Glu Phe Ile Gln Gln Ser Pro Thr Arg Ser Pro Leu Cys Leu Leu 165 170 175 tcc agt tcg tta cca tta agt tca cag ttt aaa agg cca cat cag aat 577 Ser Ser Ser Leu Pro Leu Ser Ser Gln Phe Lys Arg Pro His Gln Asn 180 185 190 acc tcc agg cct tca gag ttg ctt tta ctt gag act cct ctc ata gtt 625 Thr Ser Arg Pro Ser Glu Leu Leu Leu Leu Glu Thr Pro Leu Ile Val 195 200 205 aag aaa gct aaa tct ttg gct ctg tca gcc acg tct tct gga ttt gcc 673 Lys Lys Ala Lys Ser Leu Ala Leu Ser Ala Thr Ser Ser Gly Phe Ala 210 215 220 gag ttt act cct cca tcc atc ctt agg tct ggt ttt cga aca aca cct 721 Glu Phe Thr Pro Pro Ser Ile Leu Arg Ser Gly Phe Arg Thr Thr Pro 225 230 235 240 tta gca tct ccc tct ttg tca cct gga aga tct ctc act ccg cct ttc 769 Leu Ala Ser Pro Ser Leu Ser Pro Gly Arg Ser Leu Thr Pro Pro Phe 245 250 255 aga gtt aaa gaa aca agg att tca ttc atg gaa gaa ggc atg aat aca 817 Arg Val Lys Glu Thr Arg Ile Ser Phe Met Glu Glu Gly Met Asn Thr 260 265 270 cac tgg act gat aga gct aca gat gac cga aat aca aaa gcg ttt gtt 865 His Trp Thr Asp Arg Ala Thr Asp Asp Arg Asn Thr Lys Ala Phe Val 275 280 285 agc aca tct ttc cat aaa tgt gga ctt cca gca gaa act gag tgg atg 913 Ser Thr Ser Phe His Lys Cys Gly Leu Pro Ala Glu Thr Glu Trp Met 290 295 300 aag acc agt gat aag aat aca tat ttt cct ctg gat gtc cct gca aag 961 Lys Thr Ser Asp Lys Asn Thr Tyr Phe Pro Leu Asp Val Pro Ala Lys 305 310 315 320 ggc cct cag aaa gtg gtg gca gag tca ctg gct acc cat tca gga agg 1009 Gly Pro Gln Lys Val Val Ala Glu Ser Leu Ala Thr His Ser Gly Arg 325 330 335 ctg gag aaa ctg gat gtg agc aaa gaa gac agc aca gct tcc acc agg 1057 Leu Glu Lys Leu Asp Val Ser Lys Glu Asp Ser Thr Ala Ser Thr Arg 340 345 350 tca gac cag acc tcc tta gag tat cat gac gca cca tca cca gaa gac 1105 Ser Asp Gln Thr Ser Leu Glu Tyr His Asp Ala Pro Ser Pro Glu Asp 355 360 365 ttg gaa ggt gct gtt ttt gtg tct ccc aag cca gca tct tcc tcc act 1153 Leu Glu Gly Ala Val Phe Val Ser Pro Lys Pro Ala Ser Ser Ser Thr 370 375 380 gaa cta act act aat tca act cta caa aca gag agg gat aat gat aaa 1201 Glu Leu Thr Thr Asn Ser Thr Leu Gln Thr Glu Arg Asp Asn Asp Lys 385 390 395 400 gat gcg ttt aag tca gaa ggt act cct tca ccc gtg aag aaa caa ata 1249 Asp Ala Phe Lys Ser Glu Gly Gly Thr Pro Ser Pro Val Lys Lys Gln Ile 405 410 415 ggc acg gga gac gct gca gtg gaa gca ttt tca gaa ctg agt cgc tta 1297 Gly Thr Gly Asp Ala Ala Val Glu Ala Phe Ser Glu Leu Ser Arg Leu 420 425 430 gac cct gtt gaa aga gct gaa gct tct ttt ggt gtg tcg tca gtc tgt 5 Asp Pro Val Glu Arg Ala Glu Ala Ser Phe Gly Val Ser Ser Val Cys 435 440 445 gaa ggg gaa acc tcc act tca aac tcc aag acg tca gtt ctg gat gga 1393 Glu Gly Glu Thr Ser Thr Ser Asn Ser Lys Thr Ser Val Leu Asp Gly 450 455 460 atc gtg cct att gag agc cga acc tcc ata ctt aca gca gac cac aaa 1441 Ile Val Pro Ile Glu Ser Arg Thr Ser Ile Leu Thr Ala Asp His Lys 465 470 470 475 480 gag tct gtg gcc aac acg gtt gca gat gtt gaa agc tct ggg tcc acc 1489 Glu Ser Val Ala Asn Thr Val Ala Asp Val Glu Ser Ser Gly Ser Thr 485 490 495 agc tcc aag tgc ccg gtt acc tct gaa cgc agc ctc ggc caa aaa cta 1537 Ser Ser Lys Cys Pro Val Thr Ser Glu Arg Ser Leu Gly Gln Lys Leu 500 505 510 aca tta aac tta aaa gaa gat gaa ata gaa gct cat gta cca aag gag 1585 Thr Leu Asn Leu Lys Glu Asp Glu Ile Glu Ala His Val Pro Lys Glu 515 520 525 aac gtt ggt tta cca gaa gaa agc cct cga att tct gct gct cct tct 1633 Asn Val Gly Leu Pro Glu Glu Ser Pro Arg Ile Ser Ala Ala Ala Pro Ser 530 535 540 gat act cac gag att cat cta att gga tgt g aa aat ctt gaa gtt caa 1681 Asp Thr His Glu Ile His Leu Ile Gly Cys Glu Asn Leu Glu Val Gln 545 550 555 560 aat tca gaa gag gag gcc aag aat ctt tca ttt gat gag ttg tat ccc 1729 Asn Ser Glu Glu Glu Alu Lys Asn Leu Ser Phe Asp Glu Leu Tyr Pro 565 570 575 tta ggg gca gag aaa ctt gag tat aat ctc agt act att gag cag cag 1777 Leu Gly Ala Glu Lys Leu Glu Tyr Asn Leu Ser Thr Ile Glu Gln Gln 580 585 590 590 ttt tgt gac ttg cct gat gac aaa gac tct gct gaa tgt gat gct gct 1825 Phe Cys Asp Leu Pro Asp Asp Lys Asp Ser Ala Glu Cys Asp Ala Ala 595 600 605 gaa gta gac ggg gaa ctt ttt gtg gcc catt agc tac att 1873 Glu Val Asp Gly Glu Leu Phe Val Ala Gln Ser Asn Phe Thr Leu Ile 610 615 620 tta gaa ggt gaa gaa gga gaa gct gag gca agc gac tct gca gca cct 1921 Leu Glu Gly Glu Glu Gly Glu Ala Glu Ala Ser Ser Ala Ala Pro 625 630 635 640 aat atg tta ccg aaa tcg acc aag gaa aaa cct gtg tgc tac agg gaa 1969 Asn Met Leu Pro Lys Ser Thr Lys Glu Lys Pro Val Cys Tyr Arg Glu 645 650 655 ccc cat aat cag gag cgc gtt aca gat ttg cca tct gct gtg act gct 2017 Pro His Asn Gln Glu Arg Val Thr Asp Leu Pro Ser Ala Val Thr Ala 660 665 670 gac caa gaa tcc cac aag gta gag act tta ccg tat gtg cct gaa ccg 2065 Asp Gln Glu Ser His Lys Val Glu Thr Leu Pro Tyr Val Pro Glu Pro 675 680 685 gtt aaa gtg gca att gca gaa aat ctg ttg gat gta att aaa gac acc 2113 Val Lys Val Ala Ile Ala Glu Asn Leu Leu Asp Val Ile Lys Asp Thr 690 695 700 aga agt aag gaa gca act ccc gtg gca gca ggt gag gct ggt gat gag 2161 Arg Ser Lys Glu Ala Thr Pro Val Ala Ala Gly Glu Ala Gly Asp Glu 705 710 710 715 720 gac gga gca gtg ata gtc tca aag gct gca cat tcg tcc agg ctg aca 2209 Asp Gly Ala Val Ile Val Ser Lys Ala Ala His Ser Ser Arg Leu Thr 725 730 735 aac tct aca ccg aag act gtt aag gaa cca cgt gca gag act gta aat 2257 Asn Ser Thr Pro Lys Thr Val Lys Glu Pro Arg Ala Glu Thr Val Asn 740 745 750 acc agc cag agt gat gac atg gtt tct tct aga act ctc aca aga agg 2305 Thr Ser Gln Ser Asp Asp Met Val Ser Ser Arg Thr Leu Thr Arg Ar g 755 760 765 cag cat gcc cta agc ctg aat gtc aca tca gaa caa gag cct tca gca 2353 Gln His Ala Leu Ser Leu Asn Val Thr Ser Glu Gln Glu Pro Ser Ala 770 775 780 gtt gcc act cct aag aag aga act aga aaa att aaa gaa act cct gag 2401 Val Ala Thr Pro Lys Lys Arg Thr Arg Lys Ile Lys Glu Thr Pro Glu 785 790 795 800 tct tct gaa agg acc tgt tct gac cta aaa gta gca cct gag aac caa 2449 Ser Ser Glu Arg Thr Cys Ser Asp Leu Lys Val Ala Pro Glu Asn Gln 805 810 815 ctg aca gct cag aat cct ccc gct cct agg aga aga aag aag aag gac 2497 Leu Thr Ala Gln Asn Pro Pro Ala Pro Arg Arg Arg Lys Lys Lys Asp 820 825 830 gtt agc caa ggc aca ctg cca agt tct ggt gct gtg gag ccg gag ccg 2545 Val Ser Gln Gly Thr Leu Pro Ser Ser Gly Ala Val Glu Pro Glu Pro 835 840 845 gaa cct cag ggt acg ccg gga aga ctg agg ctg caga acg ccc 2593 Glu Pro Gln Gly Thr Pro Gly Arg Leu Arg Leu Arg Thr Gln Pro Pro 850 855 860 gag cca gca gct gaa gaa act cct tct aga aca aaa gtc agg ctt tca 2641 Glu Pro Ala Ala Glu Glu Thr Pro Ser A rg Thr Lys Val Arg Leu Ser 865 870 875 880 tct gtt aga aag gga acc cct aga aga ctt aag aag tct gta gaa aat 2689 Ser Val Arg Lys Gly Thr Pro Arg Arg Leu Lys Lys Ser Val Glu Asn 885 890 895 ggg caa agt ata gaa att cta gat gat ctc aaa ggg agt gag gca gca 2737 Gly Gln Ser Ile Glu Ile Leu Asp Asp Leu Lys Gly Ser Glu Ala Ala 900 905 910 agt cat gac ggg act gtc aca gag ctg agg aat gcc aat 785 gaat Ser His Asp Gly Thr Val Thr Glu Leu Arg Asn Ala Asn Leu Glu Asp 915 920 925 act cag aat atg gag tat aaa caa gat gaa cac agt gac cag caa ccg 2833 Thr Gln Asn Met Glu Tyr Lys Gln Asp Glu His Ser Asp Gln Gln Pro 930 935 940 cct cta aaa cga aag agg gtc aga gag aga gaa gtt agt gtg tca agt 2881 Pro Leu Lys Arg Lys Arg Val Arg Glu Arg Glu Val Ser Val Ser Ser 945 950 955 960 gtg aca gaa gag cca aag ctt gac tca tcc cag ttg cct ctt cag aca 2929 Val Thr Glu Glu Pro Lys Leu Asp Ser Ser Gln Leu Pro Leu Gln Thr 965 970 975 gga ctc gat gta cct gcc acc cct agg aaa cgt ggt aga ccc agg aag 2977 Gly Leu Asp Val Pro Ala Thr Pro Arg Lys Arg Gly Arg Pro Arg Lys 980 985 990 gta gtt ccc tta gaa gct gac ggt ggc aca act ggt aag gaa cag aca 3025 Val Val Pro Leu Glu Ala Asp Gly Gly Thr Thr Gly Lys Glu Gln Thr 995 1000 1005 agt cct cag aag aaa gat gtt ccg gtt gtc cgg aga tct aca cgg aac 3073 Ser Pro Gln Lys Lys Asp Val Pro Val Val Arg Arg Ser Thr Arg Asn 1010 1015 1020 acc cca gct aga aat gtg agt act tta naa aaa tca gtt tta gtg cca 3121 Thr Pro Ala Arg Asn Val Ser Thr Leu Xaa Lys Ser Val Leu Val Pro 1025 1030 1035 1040 aat aag aa ga gct gct cta gtg gtg aca tct aag agg aga cct aca aag 3169 Asn Lys Glu Ala Ala Leu Val Val Thr Ser Lys Arg Arg Pro Thr Lys 1045 1050 1055 aag tct gca gag gaa agc tca aaa gat cca tca gcg gca gtc tca gac 3217 Lys Ser Ala Glu Glu Ser Ser Lys Asp Pro Ser Ala Ala Val Ser Asp 1060 1065 1070 tgg gcg ggt gga gca gcc cac aca gag tcc gct gac cga agg gac gga 3265 Trp Ala Gly Gly Ala Ala His Thr Glu Ser Ala Asp Arg Arg Asp Gly 1075 1080 1085 ctg ctt gcc gcc gct gct ctc acg cca tct gcc cag ggc aca agg act 3313 Leu Leu Ala Ala Ala Ala Leu Thr Pro Ser Ala Gln Gly Thr Arg Thr 1090 1095 1100 agg tct aga agg acc atg ttg ttg acg gac att tct gaa ccc aaa act 3361 Arg Ser Arg Arg Arg Thr Met Leu Leu Thr Asp Ile Ser Glu Pro Lys Thr 1105 1110 1115 1120 gag cct tta ttt cct cct cct tca gtg aag gtt cca aag aaa aaa tca 3409 Glu Pro Leu Phe Pro Pro Ser Val Lys Val Pro Lys Lys Lys Ser 1125 1130 1135 aaa gct gag aac atg gag gcc gca gcc cag ctg aaa gaa ttg gtg tca 3457 Lys Ala Glu Asn Met Glu Ala Ala Ala Gln Leu Lys Glu Leu Val Ser 1140 1145 1150 gat tta tct tct cag ttt gtt gtt tcc cct cct aga acc agg 3505 Asp Leu Ser Ser Gln Phe Val Val Ser Pro Pro Ala Leu Arg Thr Arg 1155 1160 1165 cag aaa agt ata tcc aat act tcc aag ctt cta ggt gaa ctg gag agt 3553 Gln Lys Ser Ile Ser Asn Thr Ser Lys Leu Leu Gly Glu Leu Glu Ser 1170 1175 1180 gac cct aaa cca tta gag atc ata gaa caa aaa cca aaa aga agc agg 3601 Asp Pro Lys Pro Leu Glu Ile Ile Glu Gln Lys Pro Lys Arg Ser Arg 1185 1190 1195 1200 act gtg aag aca aga gca agc aga aac aca gga aaa gga agt tct tgg 3649 Thr Val Lys Thr Arg Ala Ser Arg Asn Thr Gly Lys Gly Ser Ser Trp 1205 1210 1215 tca cct cct cct gta gaa att aag ctg gtt tct ccc ttg gcg agt cca 3697 Ser Pro Pro Pro Val Glu Ile Lys Leu Val Ser Pro Leu Ala Ser Pro 1220 1225 1230 gtg gat gaa ata aag acc ggc aag cca aga aaa act gca gaa ata gca 3745 Val Asp Glu Ile Lys Thr Gly Lys Pro Arg Lys Thr Ala Glu Ile Ala 1235 1240 1245 gga aaa act ctt gga agg ggc aga aag aag cca tct tct ttt cca aag 3793 Gly Lys Thr Leu Gly Arg Gly Arg Lys Lys Lys Pro Ser Ser Phe Pro Lys 1250 1255 1260 caa att tta cgc agg aaa atg ctg taatttttag cccaagattt taacacgcac 3847 Gln Ile Leu Arg Arg Lys Met Leu 1265 1270 ctgtttgtaa aagtcaacag tatttgtgtg gattattaaa gtcaccaatt tggatgaaaa 3907 tactttatat aaattgtaca attttgtaag cagtaaatga gtaactccac atggagtgca 3967 gttcttgtag tgcaggcgtt ttatacgact tgatgcgttt atatcaatgt aaatatgact 4027 tatcattggg aggttaaata aactactgta aagtaaaaaa aaaaaaaaaa aaaaaaaaaa 4087 aaaaaaaaaa aaaaaaaaaa aaaaaaaa 4115 <210> 8 <211> 1272 <212> PRT <213> Mus musculus <400> 8 Gln Ile Leu Lys Asn Asn Leu Met Ser Asp Arg Asp Pro Arg Leu Arg 1 5 10 15 Glu Arg Ser Val Thr Arg Asn Ser Ile Leu Asp Gln Tyr Gly Lys Ile 20 25 30 Leu Pro Arg Val Gln Arg Lys Leu Ala Val Glu Arg Ala Lys Pro Tyr 35 40 45 His Leu Ser Thr Ser Ser Val Phe His Glu Val Ser Arg Pro Lys Pro 50 55 60 Leu Ser Ala Phe Pro Lys Lys Ala Ile Thr Gly Thr Val Leu Thr Arg 65 70 75 80 Ser Thr Phe Ile Ser Asn Val Leu Ser Lys Ile Gly Glu Val Trp Ala 85 90 95 Ser His Glu Pro Arg Asn Gly Val Ser Leu Phe Asn Ser Pro Lys Thr 100 105 110 Glu Gln Pro Ser Pro Val Val His Ser Phe Pro His Pro Glu Leu Pro 115 120 125 Glu Ala Phe Val Gly Thr Pro Ile Ser Asn Thr Ser Gln Arg Ile Ser 130 135 140 Arg Leu Leu Asp Leu Val Val His Pro Val Pro Gln Pro Ser Gln Cys 145 150 155 160 Leu Glu Phe Ile Gln Gln Ser Pro Thr Arg Ser Pro Leu Cys Leu Leu 165 170 175 Ser Ser Ser Leu Pro Leu Ser Ser Gln Phe Lys Arg Pro His Gln Asn 180 185 190 Thr Ser Arg Pro Ser Glu Leu Leu Leu Leu Glu Thr Pro Leu Ile Val 195 200 205 L ys Lys Ala Lys Ser Leu Ala Leu Ser Ala Thr Ser Ser Gly Phe Ala 210 215 220 Glu Phe Thr Pro Pro Ser Ile Leu Arg Ser Gly Phe Arg Thr Thr Pro 225 230 235 240 Leu Ala Ser Pro Ser Leu Ser Pro Gly Arg Ser Leu Thr Pro Pro Phe 245 250 255 Arg Val Lys Glu Thr Arg Ile Ser Phe Met Glu Glu Gly Met Asn Thr 260 265 270 His Trp Thr Asp Arg Ala Thr Asp Asp Arg Asn Thr Lys Ala Phe Val 275 280 285 Ser Thr Ser Phe His Lys Cys Gly Leu Pro Ala Glu Thr Glu Trp Met 290 295 300 Lys Thr Ser Asp Lys Asn Thr Tyr Phe Pro Leu Asp Val Pro Ala Lys 305 310 315 320 Gly Pro Gln Lys Val Val Ala Glu Ser Leu Ala Thr His Ser Gly Arg 325 330 335 Leu Glu Lys Leu Asp Val Ser Lys Glu Asp Ser Thr Ala Ser Thr Arg 340 345 350 Ser Asp Gln Thr Ser Leu Glu Tyr His Asp Ala Pro Ser Pro Glu Asp 355 360 365 Leu Glu Gly Ala Val Phe Val Ser Pro Lys Pro Ala Ser Ser Ser Thr 370 375 380 Glu Leu Thr Thr Asn Ser Thr Leu Gln Thr Glu Arg Asp Asn Asp Lys 385 390 395 400 Asp Ala Phe Lys Ser Glu Gly Thr Pro Ser Pro Val Lys Lys Gln Ile 405 410 415 Gly Thr Gly Asp Ala Ala Val Glu Ala Phe Ser Glu Leu Ser Arg Leu 420 425 430 Asp Pro Val Glu Arg Ala Glu Ala Ser Phe Gly Val Ser Ser Val Cys 435 440 445 Glu Gly Glu Thr Ser Thr Ser Asn Ser Lys Thr Ser Val Leu Asp Gly 450 455 460 Ile Val Pro Ile Glu Ser Arg Thr Ser Ile Leu Thr Ala Asp His Lys 465 470 475 480 Glu Ser Val Ala Asn Thr Val Ala Asp Val Glu Ser Ser Gly Ser Thr 485 490 495 Ser Ser Lys Cys Pro Val Thr Ser Glu Arg Ser Leu Gly Gln Lys Leu 500 505 510 Thr Leu Asn Leu Lys Glu Asp Glu Ile Glu Ala His Val Pro Lys Glu 515 520 525 Asn Val Gly Leu Pro Glu Glu Ser Pro Arg Ile Ser Ala Ala Pro Ser 530 535 540 Asp Thr His Glu Ile His Leu Ile Gly Cys Glu Asn Leu Glu Val Gln 545 550 555 560 Asn Ser Glu Glu Glu Ala Lys Asn Leu Ser Phe Asp Glu Leu Tyr Pro 565 570 575 Leu Gly Ala Glu Lys Leu Glu Tyr Asn Leu Ser Thr Ile Glu Gln Gln 580 585 590 Phe Cys Asp Leu Pro Asp Asp Lys Asp Ser Ala Glu Cys Asp Ala Ala 595 600 605 Glu Val Asp Gly Glu Leu Phe Val Ala Gln Ser Asn Phe Thr Leu Ile 610 615 620 Leu Glu Gly Glu Glu Gly Glu Ala Glu Ala Ser Asp Ser Ala Ala Pro 625 630 635 640 Asn Met Leu Pro Lys Ser Thr Lys Glu Lys Pro Val Cys Tyr Arg Glu 645 650 655 Pro His Asn Gln Glu Arg Val Thr Asp Leu Pro Ser Ala Val Thr Ala 660 665 670 Asp Gln Glu Ser His Lys Val Glu Thr Leu Pro Tyr Val Pro Glu Pro 675 680 685 Val Lys Val Ala Ile Ala Glu Asn Leu Leu Asp Val Ile Lys Asp Thr 690 695 700 Arg Ser Lys Glu Ala Thr Pro Val Ala Ala Gly Glu Ala Gly Asp Glu 705 710 710 720 Asp Gly Ala Val Ile Val Ser Lys Ala Ala His Ser Ser Arg Leu Thr 725 730 735 Asn Ser Thr Pro Lys Thr Val Lys Glu Pro Arg Ala Glu Thr Val Asn 740 745 750 Thr Ser Gln Ser Asp Asp Met Val Ser Ser Arg Thr Leu Thr Arg Arg 755 760 765 Gln His Ala Leu Ser Leu Asn Val Thr Ser Glu Gln Glu Pro Ser Ala 770 775 780 val Ala Thr Pro Lys Lys Arg Thr Arg Lys Ile Lys Glu Thr Pro Glu 785 790 795 800 Ser Ser Glu Arg Thr Cys Ser Asp Leu Lys Val Ala Pro Glu Asn Gln 805 810 815 Leu Thr Ala Gln Asn Pro Pro Ala Pro Arg Ar g Arg Lys Lys Lys Asp 820 825 830 Val Ser Gln Gly Thr Leu Pro Ser Ser Gly Ala Val Glu Pro Glu Pro 835 840 845 Glu Pro Gln Gly Thr Pro Gly Arg Leu Arg Leu Arg Thr Gln Pro Pro 850 855 860 Glu Pro Ala Ala Glu Glu Thr Pro Ser Arg Thr Lys Val Arg Leu Ser 865 870 875 880 Ser Val Arg Lys Gly Thr Pro Arg Arg Leu Lys Lys Ser Val Glu Asn 885 890 895 Gly Gln Ser Ile Glu Ile Leu Asp Asp Leu Lys Gly Ser Glu Ala Ala 900 905 910 Ser His Asp Gly Thr Val Thr Glu Leu Arg Asn Ala Asn Leu Glu Asp 915 920 925 Thr Gln Asn Met Glu Tyr Lys Gln Asp Glu His Ser Asp Gln Gln Pro 930 935 940 Pro Leu Lys Arg Lys Arg Val Arg Glu Arg Glu Val Ser Val Ser Ser 945 950 955 960 Val Thr Glu Glu Pro Lys Leu Asp Ser Ser Gln Leu Pro Leu Gln Thr 965 970 975 Gly Leu Asp Val Pro Ala Thr Pro Arg Lys Arg Gly Arg Pro Arg Lys 980 985 990 Val Val Pro Leu Glu Ala Asp Gly Gly Thr Thr Gly Lys Glu Gln Thr 995 1000 1005 Ser Pro Gln Lys Lys Asp Val Pro Val Val Arg Arg Ser Thr Arg Asn 1010 1015 1020 Thr Pro Ala Arg Asn Val Ser Thr Leu Xaa Lys Ser Val Leu Val Pro 1025 1030 1035 1040 Asn Lys Glu Ala Ala Leu Val Val Thr Ser Lys Arg Arg Pro Thr Lys 1045 1050 1055 Lys Ser Ala Glu Glu Ser Ser Lys Asp Pro Ser Ala Ala Val Ser Asp 1060 1065 1070 Trp Ala Gly Gly Ala Ala His Thr Glu Ser Ala Asp Arg Arg Asp Gly 1075 1080 1085 Leu Leu Ala Ala Ala Ala Leu Thr Pro Ser Ala Gln Gly Thr Arg Thr 1090 1095 1100 Arg Ser Arg Arg Thr Met Leu Leu Thr Asp Ile Ser Glu Pro Lys Thr 1105 1110 1115 1120 Glu Pro Leu Phe Pro Pro Pro Ser Val Lys Val Pro Lys Lys Lys Ser 1125 1130 1135 Lys Ala Glu Asn Met Glu Ala Ala Ala Gln Leu Lys Glu Leu Val Ser 1140 1145 1150 Asp Leu Ser Ser Gln Phe Val Val Ser Pro Pro Ala Leu Arg Thr Arg 1155 1160 1165 Gln Lys Ser Ile Ser Asn Thr Ser Lys Leu Leu Gly Glu Leu Glu Ser 1170 1175 1180 Asp Pro Lys Pro Leu Glu Ile Ile Glu Gln Lys Pro Lys Arg Ser Arg 1185 1190 1195 1200 Thr Val Lys Thr Arg Ala Ser Arg Asn Thr Gly Lys Gly Ser Ser Trp 1205 1210 1215 Ser Pro Pro Pro Val Glu Ile Lys Leu Val Ser Pro Leu Ala Ser Pro 1220 1225 1230 Val Asp Glu Ile Lys Thr Gly Lys Pro Arg Lys Thr Ala Glu Ile Ala 1235 1240 1245 Gly Lys Thr Leu Gly Arg Gly Arg Lys Lys Pro Ser Ser Phe Pro Lys 1250 1255 1260 Gln Ile Leu Arg Arg Lys Met Leu 1265 1270 <210> 9 <211> 24 <212> DNA <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Artificially Synthesized Primer Sequence <400> 9 cacccgtgaa gaaacaaata ggca 24 <210> 10 <211> 24 <212> DNA <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Artificially Synthesized Primer Sequence <400> 10 cctttggtac atgagcttct attt 24 <210> 11 <211> 7034 <212> DNA <213> Mus musculus <220> <221> CDS <222> (8) .. (6736) <400> 11 tggcagt atg caa gac ttg aca gct caa gtg act agt gat ctc ctg cat 49 Met Gln Asp Leu Thr Ala Gln Val Thr Ser Asp Leu Leu His 1 5 10 ttc cca gaa gtg act att gaa gct ctt gga gaa gat gag ata aca tta 97 Phe Pro Glu Val Thr Ile Glu Ala Leu Gly Glu Asp Glu Ile Thr Leu 15 20 25 30 gag tcc gtg ctt cgt gga aag ttt gct gca ggg aaa aat gga cta gca 145 Glu Ser Val Leu Arg Gly Lys Phe Ala Ala Gly Lys Asn Gly Leu Ala 35 40 45 tgc tta gct tgt ggt cca caa ctt gaa gtt gta aac tcc tta aca gga 193 Cys Leu Ala Cys Gly Pro Gln Leu Glu Val Val Asn Ser Leu Thr Gly 50 55 60 gag cgg tta tct gca tat aga ttc agt gga gta aat gaa cag cct cct 241 Glu Arg Leu Ser Ala Tyr Arg Phe Ser Gly Val Asn Glu Gln Pro Pro 65 70 75 gta gtc ctt gca gtg aaa gaa ttc tct tgg cat aag agg act gga ttg 289 Val Val Leu Ala Val Lys Glu Phe Ser Trp His Lys Arg Thr Gly Leu 80 85 90 tta ata gga ttg gaa gaa gca gat ggg agt gtt ctt tgt ctt tat gac 337 Leu Ile Gly Leu Glu Glu Ala Asp Gly Ser Val Leu Cys Leu Tyr Asp 95 100 105 110 ctt ggt ata tca aga gtg gtc aaa gca gtt gtt ctt cct gga agg gta 385 Leu Gly Ile Ser Arg Val Val Lys Ala Val Val Leu Pro Gly Arg Val 115 120 125 aca gct atc gag cct ata att aac cat gga gga gcc agt gcg agt acc 433 Thr Ala Ile Glu Pro Ile Ile Asn His Gly Gly Ala Ser Ala Ser Thr 130 135 140 cag cat tta cat cca agt ctc cgg tgg ctt ttt ggc gtg gcc gct gtg 481 Gln His Leu His Pro Ser Leu Arg Trp Leu Phe Gly Val Ala Ala Val 145 150 155 gtg act gat gtt gga cag atc ctt ctt att gac ctg tgt ttg gat gac 529 Val Thr Asp Val Gly Gln Ile Leu Leu Ile Asp Leu Cys Leu Asp Asp 160 165 170 ttg tcc tgc agt cag aat gaa gtt gag gca tca gac ctt gaa gtt atc 577 Leu Ser Cys Ser Gln Asn Glu Val Glu Ala Ser Asp Leu Glu Val Ile 175 180 185 190 act ggt atc cca gct gaa gta cca cac atc aga gag aga gtg atg aga 625 Thr Gly Ile Pro Ala Glu Val Pro His Ile Arg Glu Arg Val Met Arg 195 200 205 gag ggg cgc cac ctg tgc ttc cag tta gta agc cca ttg gga gta gcc 673 Glu Gly Arg His Leu Cys Phe Gln Leu Val Ser Pro Leu Gly Val Ala 210 215 220 att tct act ctg agt tac atc aac agg aca aat cag ctt gct gtg ggt 721 Ile Ser Thr Leu Ser Tyr Ile Asn Arg Thr Asn Gln Leu Ala Val Gly 225 230 235 ttt tct gat ggc tac tta gca ctt tgg aac atg aaa agc atg aaa aga 769 Phe Ser Asp Gly Tyr Leu Ala Leu Trp Asn Met Lys Ser Met Lys Arg 240 245 250 gaa tac tat aca cag ttg gaa ggt gga agg gtt cct gtc cat gca gtt 817 Glu Tyr Tyr Thr Glnu Gly Gly Arg Val Pro Val His Ala Val 255 260 265 270 gcc ttt caa gag cct gag aat gat cct cgt aac tgc tgt tat tta tgg 865 Ala Phe Gln Glu Pro Glu Asn Asp Pro Arg Asn Cys Cys Tyr Leu Trp 275 280 285 285 gct gtt cag tcc aca caa gat agt gaa ggg gat gtt ttg agt ttg cat 913 Ala Val Gln Ser Thr Gln Asp Ser Glu Gly Asp Val Leu Ser Leu His 290 295 300 ctg ctt cag ctg gct ttt ggt gat aga aaa tgt ttg gca tca ggg caa 961 Leu Leu Gln Leu Ala Phe Gly Asp Arg Lys Cys Leu Ala Ser Gly Gln 305 310 315 att tta tat gag gga tta gaa tac tgc gaa gaa aga tat aca ctg gat 1009 Ile Leu Tyr Glu Gly Leu Glu Tyr Cy Glu Arg Tyr Thr Leu Asp 320 325 330 cta gca ggt ggc acg ttc ccc tta agg gga caa act agt aat acc aaa 1057 Leu Ala Gly Gly Thr Phe Pro Leu Arg Gly Gln Thr Ser Asn Thr Lys 335 340 345 350 ttg ttg gga tgc cag agt ata gag aga ttt cca tct cat gga gac aga 1105 Leu Leu Gly Cys Gln Ser Ile Glu Arg Phe Pro Ser His Gly Asp Arg 355 360 365 gaa gaa agt atg aga gaa gct ctg tct ccc gat acc agc gtt tct gtc 1153 Glu Ser Met Arg Glu Ala Leu Ser Pro Asp Thr Ser Val Ser Val 370 375 380 ttt acc tgg caa gtg aat ata tat gga cag gga aag cct tct gtg tat 1201 Phe Thr Trp Gln Val Asn Ile Tyr Gly Gln Gly Lys Pro Ser Val Tyr 385 390 395 tta ggg cta ttt gac ata aat cgt tgg tat cat gca caa atg ccc gat 1249 Leu Gly Leu Phe Asp Ile Asn Arg Trp Tyr His Ala Gln Met Pro Asp 400 405 410 tct tta aga tca gga gaa tct ctg cat aat tgc tct tat ttt gcg ttg 1297 Ser Leu Arg Ser Gly Glu Ser Leu His Asn Cys Ser Tyr Phe Ala Leu 415 420 425 430 tgg tca ttg gat tcg gtt gta agt agg act tct cca cat cac atc ttg 1345 Trp Ser L eu Asp Ser Val Val Ser Arg Thr Ser Pro His His Ile Leu 435 440 445 gac ata cta gta cat gag agg agt tta aac cga ggg gtt cct cct tcc 1393 Asp Ile Leu Val His Glu Arg Ser Leu Asn Arg Gly Val Pro Pro Ser 450 455 460 tac cca cct cca gag caa ttt ttt aac cca agt act ttt aat ttt gat 1441 Tyr Pro Pro Pro Glu Gln Phe Phe Asn Pro Ser Thr Phe Asn Phe Asp 465 470 475 gcc act tgt ttg tta gac tct gga gtt atc cat gta act tgt gct gga 1489 Ala Thr Cys Leu Leu Asp Ser Gly Val Ile His Val Thr Cys Ala Gly 480 485 490 ttt cag aag gag act ttg aca ttt tta aag aaa tca gga cca act ctt 1537 Phe Gln Lys Glu Thr Leu Thr Phe Leu Lys Lys Ser Gly Pro Thr Leu 495 500 505 510 aat gaa gtc att cct gat agt tat aat cga tgt ctt gtt gct ggt ctc 1585 Asn Glu Val Ile Pro Asp Ser Tyr Asn Arg Cys Leu Val Ala Gly Leu 515 520 520 ctc tca cca aga ctt att gat att cag cct tcc agt tta agt caa gaa 1633 Leu Ser Pro Arg Leu Ile Asp Ile Gln Pro Ser Ser Leu Ser Gln Glu 530 535 540 gaa caa tta gaa gct ata ttg tca gca gca att cag aca agt tcc ttg 1681 Glu Gln Leu Glu Ala Ile Leu Ser Ala Ala Ile Gln Thr Ser Ser Leu 545 550 555 gga ctt ttg act ggt tac atc aga aca tgg ata ata ata gaa gaa caa cca 1729 Gly Leu Leu Thr Gly Tyr Ile Arg Trp Ile Ile Glu Glu Gln Pro 560 565 570 aat tct gct gct aat cta cga ttt gtt ctt gag tgg aca tgg aat aaa 1777 Asn Ser Ala Ala Asn Leu Arg Phe Val Leu Glu Trp Thr Trp Asn Lys 575 580 585 585 590 gtg gtt ctca aaa gaa gag ttt gat agg ctt tgt gtg ccg ctg ttt 1825 Val Val Leu Thr Lys Glu Glu Phe Asp Arg Leu Cys Val Pro Leu Phe 595 600 605 gac ggt tcg tgt cgt ttt att gac cca cag act att cag tct atc cag 1873 Asp Gly Ser Cys Arg Phe Ile Asp Pro Gln Thr Ile Gln Ser Ile Gln 610 615 620 cag tgc cat tta ctg ctt agc aac ctt agt aca gtc tta agc tgt ttt 1921 Gln Cys His Leu Leu Leu Ser Asn Leu Ser Thr Val Leu Ser Cys Phe 625 630 635 gca atg gag gcc cag ggt atc act gag aga gga ctg gtg gac ttg agc 1969 Ala Met Glu Ala Gln Gly Ile Thr Glu Arg Gly Leu Val Asp Leu Ser 640 645 650 aac aag cac atg gtc acc cag ctt ctc tgt cag tac gca cac atg gtt 2017 Asn Lys His Met Val Thr Gln Leu Leu Cys Gln Tyr Ala His Met Val 655 660 665 670 670 ctg tgg ttc tgc cac tcg ggg ctt ctg ccc gaa ggc tta gat gat gct Phe 2065 Cys His Ser Gly Leu Leu Pro Glu Gly Leu Asp Asp Ala 675 680 685 ctg cac ctg tca aga cta cgc tac aac tac cct gta att cag aac tac 2113 Leu His Leu Ser Arg Leu Arg Tyr Asn Tyr Pro Val Ile Gln Asn Tyr 690 695 700 tat aca agt cgt cgg cag aag tct gag cgc tca ccc aga ggg aag tgg 2161 Tyr Thr Ser Arg Arg Gln Lys Ser Glu Arg Ser Pro Arg Gly Lys Trp 705 710 715 aac cac gac tgc ttg atg att gat gga tta gtc tct caa cta gga gat 2209 Asn His Asp Cys Leu Met Ile Asp Gly Leu Val Ser Gln Leu Gly Asp 720 725 730 gaa gtt gag aag ttg tgg aag cgg gac gaa ggt ggc aca gga aga tac 2257 Glu Val Glu Lys Leu Trp Lys Arg Glu Gly Gly Thr Gly Arg Tyr 735 740 745 750 cct cct gct agc atc cac gca tta ctt gat ata tat tta tta gac aac 2305 Pro Pro Ala Ser Ile His Ala Leu Leu Asp Ile Tyr Leu Leu Asp Asn 755 760 7 65 att acc gaa gca agc aaa cat gct att acc att tat ttg ctg ctt gat 2353 Ile Thr Glu Ala Ser Lys His Ala Ile Thr Ile Tyr Leu Leu Leu Asp 770 775 780 att atg tat tcc ttt cca aat aaa acg gat acc ccc att gaa tct ttc 2401 Ile Met Tyr Ser Phe Pro Asn Lys Thr Asp Thr Pro Ile Glu Ser Phe 785 790 795 ccc act gcc ttt gct att tct tgg ggc caa gtt aag cta gtt caa gga 2449 Pro Thr Ala Phe Ala Ile Ser Trp Gly Gln Val Lys Leu Val Gln Gly 800 805 810 ttt tgg cta cta gat cat aat gac tat gag aat ggt tta gac ctt ctg 2497 Phe Trp Leu Leu Asp His Asn Asp Tyr Glu Asn Gly Leu Asp Leu Leu 815 820 825 830 830 ttt cac cca gtt act gca aag cct gca tcg tgg caa cat tca aag ata 2545 Phe His Pro Val Thr Ala Lys Pro Ala Ser Trp Gln His Ser Lys Ile 835 840 845 att gaa gct ttt atg agt cag gga gag cac aaa cag gct ctc cgg tat 2593 Ile Glu Ala Phe Met Ser Gln Gly Glu His Lys Gln Ala Leu Arg Tyr 850 855 860 ctt cag aca atg aag cca aca gtg tcc agt agc aat gaa gtt atc ctt 2641 Leu Gln Thr Met Lys Pro Thr Val Ser Ser Ser As n Glu Val Ile Leu 865 870 875 cac ctc act gtt cta ctt ttt aat aga tgc atg gtt gag gcc tgg aac 2689 His Leu Thr Val Leu Leu Phe Asn Arg Cys Met Val Glu Ala Trp Asn 880 885 890 tta ctg cga cag aat tca aac aga gta aat ata gag gaa tta tta aag 2737 Leu Leu Arg Gln Asn Ser Asn Arg Val Asn Ile Glu Glu Leu Leu Lys 895 900 905 910 cac gct tat gaa gtt tgt cag gag atg ggc tta atg gag gat Atta latg785 Tyr Glu Val Cys Gln Glu Met Gly Leu Met Glu Asp Leu Leu 915 920 925 aag ctg cca ttt aca aac act gag cag gaa tgc tta gtg aaa ttt tta 2833 Lys Leu Pro Phe Thr Asn Thr Glu Gln Glu Cys Leu Val Lys Phe Le 930 935 940 cag tcc agt acc agt gtt gag aat cat gaa ttc ctt cta gtt cac cat 2881 Gln Ser Ser Thr Ser Val Glu Asn His Glu Phe Leu Leu Val His His 945 950 955 tta cag cgt gcc aat tat att tct gcc ttg aaa cta aac cag att ctg 2929 Leu Gln Arg Ala Asn Tyr Ile Ser Ala Leu Lys Leu Asn Gln Ile Leu 960 965 970 aag aat aat ctc atg agt gat cgt gac cct cga ttg cgg gaa aga tcg 2977 Lys Asn Asn Leu Met Ser Asp Arg Asp Pro Arg Leu Arg Glu Arg Ser 975 980 985 990 gtg act cga aat tct ata tta gac cag tat ggg aaa atc cta cct aga 3025 Val Thr Arg Asn Ser Ile Leu Asp Gln Tyr Gly Lys Ile Leu Pro Arg 995 1000 1005 gtc cag aga aag tta gct gtt gag cga gct aag cct tac cac ctg tcg 3073 Val Gln Arg Lys Leu Ala Val Glu Arg Ala Lys Pro Tyr His Leu Ser 1010 1015 1020 aca tcc tca gtt ttt cat gaa gtt tct aga ccc aaa tta tcg gca 3121 Thr Ser Ser Val Phe His Glu Val Ser Arg Pro Lys Pro Leu Ser Ala 1025 1030 1035 ttt cca aag aaa gct ata act gga aca gtg tta acc cga tct acg ttc 3169 Phe Pro Lys Lys Ala Ile Thr Gly Thr Val Leu Thr Arg Ser Thr Phe 1040 1045 1050 atc agc aat gtt tta tct aaa att gga gag gtg tgg gca agt cat gag 3217 Ile Ser Asn Val Leu Ser Lys Ile Gly Glu Val Trp Ala Ser His Glu 1055 1060 1065 1070 cct aga aat ggc gtc tca ctt ttt aac agt cct aaa aca gaa cag cca 3265 Pro Arg Asn Gly Val Ser Leu Phe Asn Ser Pro Lys Thr Glu Gln Pro 1075 1080 1085 tct cct gta gta cac tct ttc cca cac cca g ag ctt cct gag gcg ttt 3313 Ser Pro Val Val His Ser Phe Pro His Pro Glu Leu Pro Glu Ala Phe 1090 1095 1100 gtt gga act cca att tca aat aca tcc cag aga att tct aga tta ctg 3361 Val Gly Thr Pro Ile Ser Asn Thr Ser Gln Arg Ile Ser Arg Leu Leu 1105 1110 1115 gat ttg gtt gtc cat cct gta ccc cag cct tct cag tgt ttg gag ttt 3409 Asp Leu Val Val His Pro Val Pro Gln Pro Ser Gln Cys Leu Glu Phe 1120 1125 1130 att caa caa agt ccc aca aga tct cct ttg tgt ctg ctg tcc agt tcg 3457 Ile Gln Gln Ser Pro Thr Arg Ser Pro Leu Cys Leu Leu Ser Ser Ser 1135 1140 1145 1150 tta cca tta agt tca cag ttt aaa agg cca cat cag aat acc tcc agg 3505 Leu Pro Leu Ser Ser Gln Phe Lys Arg Pro His Gln Asn Thr Ser Arg 1155 1160 1165 cct tca gag ttg ctt tta ctt gag act cct ctc ata gtt aag aaa gct 3553 Pro Ser Glu Leu Leu Leu Leu Glu Thr Pro Leu Ile Val Lys Lys Ala 1170 1175 1180 aaa tct ttg gct ctg tca gcc acg tct tct gga ttt gcc gag ttt act 3601 Lys Ser Leu Ala Leu Ser Ala Thr Ser Ser Gly Phe Ala Glu Phe Thr 1185 1190 1195 cct cca tcc atc ctt agg tct ggt ttt cga aca aca cct tta gca tct 3649 Pro Pro Ser Ile Leu Arg Ser Gly Phe Arg Thr Thr Pro Leu Ala Ser 1200 1205 1210 ccc tct ttg tca cct gga aga tct ctc act ccg cct ttc aga gtt aaa 3697 Pro Ser Leu Ser Pro Gly Arg Ser Leu Thr Pro Pro Phe Arg Val Lys 1215 1220 1225 1230 gaa aca agg att tca ttc atg gaa gaa ggc atg aat aca cac tgg act 3745 Glu Thr Arg Ile Ser Phe Met Glu Glu Gly Met Asn Thr His Trp Thr 1235 1240 1245 gat aga gct aca gat gac cga aat aca aaa gcg ttt gtt agc aca tct 3793 Asp Arg Ala Thr Asp Asp Arg Asn Thr Lys Ala Phe Val Ser Thr Ser 1250 1255 1260 ttc cat aaa tgt gga ctt cca gca gaa act gag tgg atg aag acc agt 3841 Phe His Lys Cys Gly Leu Pro Ala Glu Thr Glu Trp Met Lys Thr Ser 1265 1270 1275 gat aag aat aca tat ttt cct ctg gat gtc cct gca aag ggc cct cag 3889 Asp Lys Asn Thr Tyr Phe Pro Leu Asp Val Pro Ala Lys Gly Pro Gln 1280 1285 1290 aaa gtg gtg gca gag tca ctg gct acc cat tca gga agg ctg gag aaa 3937 Lys Val Val Ala Glu Ser L eu Ala Thr His Ser Gly Arg Leu Glu Lys 1295 1300 1305 1310 ctg gat gtg agc aaa gaa gac agc aca gct tcc acc agg tca gac cag 3985 Leu Asp Val Ser Lys Glu Asp Ser Thr Ala Ser Thr Arg Ser Asp Gln 1315 1320 1325 acc tcc tta gag tat cat gac gca cca tca cca gaa gac ttg gaa ggt 4033 Thr Ser Leu Glu Tyr His Asp Ala Pro Ser Pro Glu Asp Leu Glu Gly 1330 1335 1340 gct gtt ttt gtg tct ccc aag cca gca tct tcc tcc act gaa cta act 4081 Ala Val Phe Val Ser Pro Lys Pro Ala Ser Ser Ser Thr Glu Leu Thr 1345 1350 1355 act aat tca act cta caa aca gag agg gat aat gat aaa gat gcg ttt 4129 Thr Asn Ser Thr Leu Gln Thr Glu Arg Asp Asn Asp Lys Asp Ala Phe 1360 1365 1370 aag tca gaa ggt act cct tca ccc gtg aag aaa caa ata ggc acg gga 4177 Lys Ser Glu Gly Thr Pro Ser Pro Val Lys Lys Gln Ile Gly Thrhr Gly 1375 1380 1385 1390 gac gct gca gtg gaa gca ttt tca gaa ctg agt cgc tta gac cct gtt 4225 Asp Ala Ala Val Glu Ala Phe Ser Glu Leu Ser Arg Leu Asp Pro Val 1395 1400 1405 gaa aga gct gaa gct tct ttt ggt gtg tcg t ca gtc tgt gaa ggg gaa 4273 Glu Arg Ala Glu Ala Ser Phe Gly Val Ser Ser Val Cys Glu Gly Glu 1410 1415 1420 acc tcc act tca aac tcc aag acg tca gtt ctg gat gga atc gtg cct 4321 Thr Ser Thr Ser Asn Ser Lys Thr Ser Val Leu Asp Gly Ile Val Pro 1425 1430 1435 att gag agc cga acc tcc ata ctt aca gca gac cac aaa gag tct gtg 4369 Ile Glu Ser Arg Thr Ser Ile Leu Thr Ala Asp His Lys Glu Ser Val 1440 1445 1450 1450 gcc aac acg gtt gca gat gtt gaa agc tct ggg tcc acc agc tcc aag 4417 Ala Asn Thr Val Ala Asp Val Glu Ser Ser Gly Ser Thr Ser Ser Lys 1455 1460 1465 1470 tgc ccg gtt acc tct gaa cgc agc ctc ggc caa tta ctaca aca aac 4465 Cys Pro Val Thr Ser Glu Arg Ser Leu Gly Gln Lys Leu Thr Leu Asn 1475 1480 1485 tta aaa gaa gat gaa ata gaa gct cat gta cca aag gag aac gtt ggt 4513 Leu Lys Glu Asp Glu Ile Glu Ala His Val Pro Lys Glu Asn Val Gly 1490 1495 1500 tta cca gaa gaa agc cct cga att tct gct gct cct tct gat act cac 4561 Leu Pro Glu Glu Ser Pro Arg Ile Ser Ala Ala Pro Ser Asp Thr His 1505 1510 1515 gag att cat cta att gga tgt gaa aat ctt gaa gtt caa aat tca gaa 4609 Glu Ile His Leu Ile Gly Cys Glu Asn Leu Glu Val Gln Asn Ser Glu 1520 1525 1530 gag gag gcc aag aat ctt tca ttt gat cag ttg tta ggg gca 4657 Glu Glu Ala Lys Asn Leu Ser Phe Asp Glu Leu Tyr Pro Leu Gly Ala 1535 1540 1545 1550 gag aaa ctt gag tat aat ctc agt act att gag cag cag ttt tgt gac 4705 Glu Lys Leu Glu Tyr Asn Leu Ser Thr Ile Glu Gln Gln Phe Cys Asp 1555 1560 1565 ttg cct gat gac aaa gac tct gct gaa tgt gat gct gct gaa gta gac 4753 Leu Pro Asp Asp Lys Asp Ser Ala Glu Cys Asp Ala Ala Glu Val Asp 1570 1575 cgg gtt gaa gtg gcc cag agc aac ttt acc ctg att tta gaa ggt 4801 Gly Glu Leu Phe Val Ala Gln Ser Asn Phe Thr Leu Ile Leu Glu Gly 1585 1590 1595 gaa gaa gga gaa gct gag gca agc gac tct gca gca cct aatlug Glu Gly Glu Ala Glu Ala Ser Asp Ser Ala Ala Pro Asn Met Leu 1600 1605 1610 ccg aaa tcg acc aag gaa aaa cct gtg tgc tac agg gaa ccc cat aat 4897 Pro Lys Ser Thr Lys Glu Ly s Pro Val Cys Tyr Arg Glu Pro His Asn 1615 1620 1625 1630 cag gag cgc gtt aca gat ttg cca tct gct gtg act gct gac caa gaa 4945 Gln Glu Arg Val Thr Asp Leu Pro Ser Ala Val Thr Ala Asp Gln Glu 1635 1640 1645 tcc cac aag gta gag act tta ccg tat gtg cct gaa ccg gtt aaa gtg 4993 Ser His Lys Val Glu Thr Leu Pro Tyr Val Pro Glu Pro Val Lys Val 1650 1655 1660 gca att gca gaa aat ctg ttg gat gta att aaa gac accaga agt aag 5041 Ala Ile Ala Glu Asn Leu Leu Asp Val Ile Lys Asp Thr Arg Ser Lys 1665 1670 1675 gaa gca act ccc gtg gca gca ggt gag gct ggt gat gag gac gga gca 5089 Glu Ala Thr Pro Val Ala Ala Gly Glu Ala Gly Asp Glu Asp Gly Ala 1680 1685 1690 gtg ata gtc tca aag gct gca cat tcg tcc agg ctg aca aac tct aca 5137 Val Ile Val Ser Lys Ala Ala His Ser Ser Arg Leu Thr Asn Ser Thr 1695 1700 1705 1710 ccg aag act gtt aag gaa cca cgt gca gag act gta aat acc agc cag 5185 Pro Lys Thr Val Lys Glu Pro Arg Ala Glu Thr Val Asn Thr Ser Gln 1715 1720 1725 agt gat gac atg gtt tct tct aga act ctc ac a aga agg cag cat gcc 5233 Ser Asp Asp Met Val Ser Ser Arg Thr Leu Thr Arg Arg Gln His Ala 1730 1735 1740 cta agc ctg aat gtc aca tca gaa caa gag cct tca gca gtt gcc act 5281 Leu Ser Leu Asn Val Thr Ser Glu Gln Glu Pro Ser Ala Val Ala Thr 1745 1750 1755 cct aag aag aga act aga aaa att aaa gaa act cct gag tct tct gaa 5329 Pro Lys Lys Arg Thr Arg Lys Ile Lys Glu Thr Pro Glu Ser Ser Glu 1760 1765 1770 agg acc tgt tct gac cta aaa gta gca cct gag aac caa ctg aca gct 5377 Arg Thr Cys Ser Asp Leu Lys Val Ala Pro Glu Asn Gln Leu Thr Ala 1775 1780 1785 1790 cag aat cct ccc gct cct agg aga aga aag aag aag gac gtt ag caa 5425 Gln Asn Pro Pro Ala Pro Arg Arg Arg Lys Lys Lys Asp Val Ser Gln 1795 1800 1805 ggc aca ctg cca agt tct ggt gct gtg gag ccg gag ccg gaa cct cag 5473 Gly Thr Leu Pro Ser Ser Gly Ala Val Glu Pro Glu Pro Glu Pro Gln 1810 1815 1820 ggt acg ccg gga aga ctg agg ctg aga acg cag cca ccc gag cca gca 5521 Gly Thr Pro Gly Arg Leu Arg Leu Arg Thr Gln Pro Pro Glu Pro Ala 1825 1830 1835 gct gaa gaa act cct tct aga aca aaa gtc agg ctt tca tct gtt aga 5569 Ala Glu Glu Thr Pro Ser Arg Thr Lys Val Arg Leu Ser Ser Val Arg 1840 1845 1850 aag gga acc cct aga aga ctt aag aag tct gta gaa aat ggg caa agt 5617 Lys Gly Thr Pro Arg Arg Leu Lys Lys Ser Val Glu Asn Gly Gln Ser 1855 1860 1865 1870 ata gaa att cta gat gat ctc aaa ggg agt gag gca gca agt cat gac 5665 Ile Glu Ile Leu Asp Asp Leu Lys Gly Ser Glu Ala Ala Ser His Asp 1875 1880 1885 ggg act gtc aca gag ctg agg aat gcc aat tta gaa gat act cag aat 5713 Gly Thr Val Thr Glu Leu Arg Asn Ala Asn Leu Glu Asp Thr Gln Asn 1890 1895 1900 atg gag tat aaa caa gat gaa cac agt gac cag caa ccg cct cta aaa 5761 Met Glu Tyr Lys Gln Asp Glu His Ser Asp Gln Gln Pro Pro Leu Lys 1905 1910 1915 cga aag agg gtc aga gag aga gaa gtt agt gtg tca agt gtg aca gag Lys Arg Val Arg Glu Arg Glu Val Ser Val Ser Ser Val Thr Glu 1920 1925 1930 gag cca aag ctt gac tca tcc cag ttg cct ctt cag aca gga ctc gat 5857 Glu Pro Lys Leu Asp Ser Se r Gln Leu Pro Leu Gln Thr Gly Leu Asp 1935 1940 1945 1950 gta cct gcc acc cct agg aaa cgt ggt aga ccc agg aag gta gtt ccc 5905 Val Pro Ala Thr Pro Arg Lys Arg Gly Arg Pro Arg Lys Val Val Pro 1955 1960 1965 tta gaa gct gac ggt ggc aca act ggt aag gaa cag aca agt cct cag 5953 Leu Glu Ala Asp Gly Gly Thr Thr Gly Lys Glu Gln Thr Ser Pro Gln 1970 1975 1980 aag aaa gat gtt ccg gtt gtc cgg aga tct aca cgg aac acc cca gct 6001 Lys Lys Asp Val Pro Val Val Arg Arg Ser Thr Arg Asn Thr Pro Ala 1985 1990 1995 aga aat gtg agt act tta aaa aaa tca gtt tta gtg cca aat aag gaa 6049 Arg Asn Val Ser Thr Leu Lys Lys Ser Val Leu Val Pro Asn Lys Glu 2000 2005 2010 gct gct cta gtg gtg aca tct aag agg aga cct aca aag aag tct gca 6097 Ala Ala Leu Val Val Thr Ser Lys Arg Arg Pro Thr Lys Lys Ser Ala 2015 2020 2025 2030 gag gaa agc tca aaa gat cca tca gcg gca gtc tca gac tgg gcg ggt 6145 Glu Glu Ser Ser Lys Asp Pro Ser Ala Ala Val Ser Asp Trp Ala Gly 2035 2040 2045 gga gca gcc cac aca gag tcc gct gac cga agg gac gga ctg ctt gcc 6193 Gly Ala Ala His Thr Glu Ser Ala Asp Arg Arg Asp Gly Leu Leu Ala 2050 2055 2060 gcc gct gct ctc acg cca tct gcc cag ggc aca agg act agg tct aga 6241 Ala Ala Ala Leu Thr Pro Ser Ala Gln Gly Thr Arg Thr Arg Ser Arg 2065 2070 2075 agg acc atg ttg ttg acg gac att tct gaa ccc aaa act gag cct tta 6289 Arg Thr Met Leu Leu Leu Thr Asp Ile Ser Glu Pro Lys Thr Glu Pro Leu 2080 2085 2090 ttt cct cct cct tca gtg aag gtt cca aag aaa aaa tca aaa gct gag 6337 Phe Pro Pro Pro Ser Val Lys Val Pro Lys Lys Lys Ser Lys Ala Glu 2095 2100 2105 2110 aac atg gag gcc gca gcc cag ctg aaa gaa ttg gtg tca gat ttat 6385 Asn Met Glu Ala Ala Ala Gln Leu Lys Glu Leu Val Ser Asp Leu Ser 2 115 2120 2125 tct cag ttt gtt gtt tcc cct cct gcc ttg aga acc agg cag aaa agt 6433 Ser Gln Phe Val Val Ser Pro Pro Ala Leu Arg Thr Arg Gln Lys Ser 2130 2135 2140 ata tcc aat act tcc aag ctt cta ggt gaa ctg gag agt gac cct aaa 6481 Ile Ser Asn Thr Ser Lys Leu Leu Gly Glu Leu Glu Ser Asp Pro Lys 2145 2150 2 155 cca tta gag atc ata gaa caa aaa cca aaa aga agc agg act gtg aag 6529 Pro Leu Glu Ile Ile Glu Gln Lys Pro Lys Arg Ser Arg Thr Val Lys 2160 2165 2170 aca aga gca agc aga aac aca gga aaa gga agt tct tca cct cct 6577 Thr Arg Ala Ser Arg Asn Thr Gly Lys Gly Ser Ser Trp Ser Pro Pro 2175 2180 2185 2190 cct gta gaa att aag ctg gtt tct ccc ttg gcg agt cca gtg gat gaa 6625 Pro Val Glu Ile Lys Leu Val Ser Pro Leu Ala Ser Pro Val Asp Glu 2195 2200 2205 ata aag acc ggc aag cca aga aaa act gca gaa ata gca gga aaa act 6673 Ile Lys Thr Gly Lys Pro Arg Lys Thr Ala Glu Ile Ala Gly Lys Thr 2210 2215 2220 ctt gga agg ggc aga aag aag cca tct tct ttt cca aag caa att tta 6721 Leu Gly Arg Gly Arg Lys Lys Pro Ser Ser Phe Pro Lys Gln Ile Leu 2225 2230 2235 cgc agg aaa atg ctg taatttttag cccaagattt taacacgcac ctgtttgg agat gtgtttgaa 6gt gattattaaa gtcaccaatt tggatgaaaa tactttatat 6836 aaattgtaca attttgtaag cagtaaatga gtaactccac atggagtgca gttcttgtag 6896 tgcagg cgtt ttatacgact tgatgcgttt atatcaatgt aaatatgact tatcattggg 6956 aggttaaata aactactgta aagtaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa 7016 aaaaaaaaaa aaaaaaaa 7034 <210> 12 <211> 2243 <212> PRT <213> Mus musculus <400> 12 Met Gln Asp Leu Thr Ala Gln Val Thr Ser Asp Leu Leu His Phe Pro 1 5 10 15 Glu Val Thr Ile Glu Ala Leu Gly Glu Asp Glu Ile Thr Leu Glu Ser 20 25 30 Val Leu Arg Gly Lys Phe Ala Ala Gly Lys Asn Gly Leu Ala Cys Leu 35 40 45 Ala Cys Gly Pro Gln Leu Glu Val Val Asn Ser Leu Thr Gly Glu Arg 50 55 60 Leu Ser Ala Tyr Arg Phe Ser Gly Val Asn Glu Gln Pro Pro Val Val 65 70 75 80 Leu Ala Val Lys Glu Phe Ser Trp His Lys Arg Thr Gly Leu Leu Ile 85 90 95 Gly Leu Glu Glu Ala Asp Gly Ser Val Leu Cys Leu Tyr Asp Leu Gly 100 105 110 Ile Ser Arg Val Val Lys Ala Val Val Leu Pro Gly Arg Val Thr Ala 115 120 125 Ile Glu Pro Ile Ile Asn His Gly Gly Ala Ser Ala Ser Thr Gln His 130 135 140 Leu His Pro Ser Leu Arg Trp Leu Phe Gly Val Ala Ala Val Val Thr 145 150 155 160 Asp Val Gly Gln Ile Leu Leu Ile Asp Leu Cys Leu Asp Asp Leu Ser 165 170 175 Cys Ser Gln Asn Glu Val Glu Ala Ser Asp Leu Glu Val Ile Thr Gly 180 185 190 Ile Pro Ala Glu Val Pro His Ile Arg Glu Arg Val Met Arg Glu Gly 195 200 205 Arg His Leu Cys Phe Gln Leu Val Ser Pro Leu Gly Val Ala Ile Ser 210 215 220 Thr Leu Ser Tyr Ile Asn Arg Thr Asn Gln Leu Ala Val Gly Phe Ser 225 230 235 240 Asp Gly Tyr Leu Ala Leu Trp Asn Met Lys Ser Met Lys Arg Glu Tyr 245 250 255 Tyr Thr Gln Leu Glu Gly Gly Arg Val Pro Val His Ala Val Ala Phe 260 265 270 Gln Glu Pro Glu Asn Asp Pro Arg Asn Cys Cys Tyr Leu Trp Ala Val 275 280 285 Gln Ser Thr Gln Asp Ser Glu Gly Asp Val Leu Ser Leu His Leu Leu 290 295 300 Gln Leu Ala Phe Gly Asp Arg Lys Cys Leu Ala Ser Gly Gln Ile Leu 305 310 315 320 Tyr Glu Gly Leu Glu Tyr Cys Glu Glu Arg Tyr Thr Leu Asp Leu Ala 325 330 335 Gly Gly Thr Phe Pro Leu Arg Gly Gln Thr Ser Asn Thr Lys Leu Leu 340 345 350 Gly Cys Gln Ser Ile Glu Arg Phe Pro Ser His Gly Asp Arg Glu Glu 355 360 365 365 Ser Met Arg Glu Ala Leu Ser Pro Asp Thr Ser Val Ser Val Phe Thr 370 375 380 Trp Gln Val Asn Ile Tyr Gly Gln Gly Lys Pro Ser Val Tyr Leu Gly 385 390 395 400 Leu Phe Asp Ile Asn Arg Trp Tyr His Ala Gln Met Pro Asp Ser Leu 4 05 410 415 Arg Ser Gly Glu Ser Leu His Asn Cys Ser Tyr Phe Ala Leu Trp Ser 420 425 430 Leu Asp Ser Val Val Ser Arg Thr Ser Pro His His Ile Leu Asp Ile 435 440 445 Leu Val His Glu Arg Ser Leu Asn Arg Gly Val Pro Pro Ser Tyr Pro 450 455 460 Pro Pro Glu Gln Phe Phe Asn Pro Ser Thr Phe Asn Phe Asp Ala Thr 465 470 475 480 Cys Leu Leu Asp Ser Gly Val Ile His Val Thr Cys Ala Gly Phe Gln 485 490 495 Lys Glu Thr Leu Thr Phe Leu Lys Lys Ser Gly Pro Thr Leu Asn Glu 500 505 510 Val Ile Pro Asp Ser Tyr Asn Arg Cys Leu Val Ala Gly Leu Leu Ser 515 520 525 Pro Arg Leu Ile Asp Ile Gln Pro Ser Ser Leu Ser Gln Glu Glu Gln 530 535 540 Leu Glu Ala Ile Leu Ser Ala Ala Ile Gln Thr Ser Ser Leu Gly Leu 545 550 555 560 Leu Thr Gly Tyr Ile Arg Thr Trp Ile Ile Glu Glu Gln Pro Asn Ser 565 570 575 Ala Ala Asn Leu Arg Phe Val Leu Glu Trp Thr Trp Asn Lys Val Val 580 585 590 Leu Thr Lys Glu Glu Phe Asp Arg Leu Cys Val Pro Leu Phe Asp Gly 595 600 605 Ser Cys Arg Phe Ile Asp Pro Gln Thr Ile Gln Ser Ile Gln Gln Cys 610 615 620 His Leu Leu Leu Ser Asn Leu Ser Thr Val Leu Ser Cys Phe Ala Met 625 630 635 640 Glu Ala Gln Gly Ile Thr Glu Arg Gly Leu Val Asp Leu Ser Asn Lys 645 650 655 His Met Val Thr Gln Leu Leu Cys Gln Tyr Ala His Met Val Leu Trp 660 665 670 Phe Cys His Ser Gly Leu Leu Pro Glu Gly Leu Asp Asp Ala Leu His 675 680 685 Leu Ser Arg Leu Arg Tyr Asn Tyr Pro Val Ile Gln Asn Tyr Tyr Thr 690 695 700 Ser Arg Arg Gln Lys Ser Glu Arg Ser Pro Arg Gly Lys Trp Asn His 705 710 715 720 Asp Cys Leu Met Ile Asp Gly Leu Val Ser Gln Leu Gly Asp Glu Val 725 730 735 Glu Lys Leu Trp Lys Arg Asp Glu Gly Gly Thr Gly Arg Tyr Pro Pro 740 745 750 Ala Ser Ile His Ala Leu Leu Asp Ile Tyr Leu Leu Asp Asn Ile Thr 755 760 765 Glu Ala Ser Lys His Ala Ile Thr Ile Tyr Leu Leu Leu Asp Ile Met 770 775 775 780 Tyr Ser Phe Pro Asn Lys Thr Asp Thr Pro Ile Glu Ser Phe Pro Thr 785 790 795 800 Ala Phe Ala Ile Ser Trp Gly Gln Val Lys Leu Val Gln Gly Phe Trp 805 810 815 Leu Leu Asp His Asn Asp Tyr Glu Asn Gly Leu Asp Leu Leu Phe His 820 825 830 Pro Val Thr Ala Lys Pro Ala Ser Trp Gln His Ser Lys Ile Ile Glu 835 840 845 Ala Phe Met Ser Gln Gly Glu His Lys Gln Ala Leu Arg Tyr Leu Gln 850 855 860 Thr Met Lys Pro Thr Val Ser Ser Ser Asn Glu Val Ile Leu His Leu 865 870 875 880 880 Thr Val Leu Leu Phe Asn Arg Cys Met Val Glu Ala Trp Asn Leu Leu 885 890 895 Arg Gln Asn Ser Asn Arg Val Asn Ile Glu Glu Leu Leu Lys His Ala 900 905 910 Tyr Glu Val Cys Gln Glu Met Gly Leu Met Glu Asp Leu Leu Lys Leu 915 920 925 Pro Phe Thr Asn Thr Glu Gln Glu Cys Leu Val Lys Phe Leu Gln Ser 930 935 940 940 Ser Thr Ser Val Glu Asn His Glu Phe Leu Leu Val His His Leu Gln 945 950 955 960 Arg Ala Asn Tyr Ile Ser Ala Leu Lys Leu Asn Gln Ile Leu Lys Asn 965 970 975 Asn Leu Met Ser Asp Arg Asp Pro Arg Leu Arg Glu Arg Ser Val Thr 980 985 990 Arg Asn Ser Ile Leu Asp Gln Tyr Gly Lys Ile Leu Pro Arg Val Gln 995 1000 1005 Arg Lys Leu Ala Val Glu Arg Ala Lys Pro Tyr His Leu Ser Thr Ser 1010 1015 1020 Ser Val Phe His Glu Val Ser Ar g Pro Lys Pro Leu Ser Ala Phe Pro 1025 1030 1035 1040 Lys Lys Ala Ile Thr Gly Thr Val Leu Thr Arg Ser Thr Phe Ile Ser 1045 1050 1055 Asn Val Leu Ser Lys Ile Gly Glu Val Trp Ala Ser His Glu Pro Arg 1060 1065 1070 Asn Gly Val Ser Leu Phe Asn Ser Pro Lys Thr Glu Gln Pro Ser Pro 1075 1080 1085 Val Val His Ser Phe Pro His Pro Glu Leu Pro Glu Ala Phe Val Gly 1090 1095 1100 Thr Pro Ile Ser Asn Thr Ser Gln Arg Ile Ser Arg Leu Leu Asp Leu 1105 1110 1115 1120 Val Val His Pro Val Pro Gln Pro Ser Gln Cys Leu Glu Phe Ile Gln 1125 1130 1135 Gln Ser Pro Thr Arg Ser Pro Leu Cys Leu Leu Ser Ser Ser Leu Pro 1140 1145 1150 Leu Ser Ser Gln Phe Lys Arg Pro His Gln Asn Thr Ser Arg Pro Ser 1155 1160 1165 Glu Leu Leu Leu Leu Glu Thr Pro Leu Ile Val Lys Lys Ala Lys Ser 1170 1175 1180 Leu Ala Leu Ser Ala Thr Ser Ser Gly Phe Ala Glu Phe Thr Pro Pro 1185 1190 1195 1200 Ser Ile Leu Arg Ser Gly Phe Arg Thr Thr Pro Leu Ala Ser Pro Ser 1205 1210 1215 Leu Ser Pro Gly Arg Ser Leu Thr Pro Pro Phe Arg Val Lys Glu Thr 1220 1225 1230 Arg Ile Ser Phe Met Glu Glu Gly Met Asn Thr His Trp Thr Asp Arg 1235 1240 1245 Ala Thr Asp Asp Arg Asn Thr Lys Ala Phe Val Ser Thr Ser Phe His 1250 1255 1260 Lys Cys Gly Leu Pro Ala Glu Thr Glu Trp Met Lys Thr Ser Asp Lys 1265 1270 1275 1280 Asn Thr Tyr Phe Pro Leu Asp Val Pro Ala Lys Gly Pro Gln Lys Val 1285 1290 1295 Val Ala Glu Ser Leu Ala Thr His Ser Gly Arg Leu Glu Lys Leu Asp 1300 1305 1310 Val Ser Lys Glu Asp Ser Thr Ala Ser Thr Arg Ser Asp Gln Thr Ser 1315 1320 1325 Leu Glu Tyr His Asp Ala Pro Ser Pro Glu Asp Leu Glu Gly Ala Val 1330 1335 1340 Phe Val Ser Pro Lys Pro Ala Ser Ser Ser Thr Glu Leu Thr Thr Asn 1345 1350 1355 1360 Ser Thr Leu Gln Thr Glu Arg Asp Asn Asp Lys Asp Ala Phe Lys Ser 1365 1370 1375 Glu Gly Thr Pro Ser Pro Val Lys Lys Gln Ile Gly Thr Gly Asp Ala 1380 1385 1390 Ala Val Glu Ala Phe Ser Glu Leu Ser Arg Leu Asp Pro Val Glu Arg 1395 1400 1405 Ala Glu Ala Ser Phe Gly Val Ser Ser Val Cys Glu Gly Glu Thr Ser 1410 1415 1420 Thr Se r Asn Ser Lys Thr Ser Val Leu Asp Gly Ile Val Pro Ile Glu 1425 1430 1435 1440 Ser Arg Thr Ser Ile Leu Thr Ala Asp His Lys Glu Ser Val Ala Asn 1445 1450 1455 Thr Val Ala Asp Val Glu Ser Ser Gly Ser Thr Ser Ser Lys Cys Pro 1460 1465 1470 Val Thr Ser Glu Arg Ser Leu Gly Gln Lys Leu Thr Leu Asn Leu Lys 1475 1480 1485 Glu Asp Glu Ile Glu Ala His Val Pro Lys Glu Asn Val Gly Leu Pro 1490 1495 1500 Glu Glu Ser Pro Arg Ile Ser Ala Ala Pro Ser Asp Thr His Glu Ile 1505 1510 1515 1520 His Leu Ile Gly Cys Glu Asn Leu Glu Val Gln Asn Ser Glu Glu Glu 1525 1530 1535 Ala Lys Asn Leu Ser Phe Asp Glu Leu Tyr Pro Leu Gly Ala Glu Lys 1540 1545 1550 Leu Glu Tyr Asn Leu Ser Thr Ile Glu Gln Gln Phe Cys Asp Leu Pro 1555 1560 1565 Asp Asp Lys Asp Ser Ala Glu Cys Asp Ala Ala Ala Glu Val Asp Gly Glu 1570 1575 1580 Leu Phe Val Ala Gln Ser Asn Phe Thr Leu Ile Leu Glu Gly Glu Glu 1585 1590 1595 1600 Gly Glu Ala Glu Ala Ser Asp Ser Ala Ala Pro Asn Met Leu Pro Lys 1605 1610 1615 Ser Thr Lys Glu Lys Pro Val Cys Tyr Arg Glu Pro His Asn Gln Glu 1620 1625 1630 Arg Val Thr Asp Leu Pro Ser Ala Val Thr Ala Asp Gln Glu Ser His 1635 1640 1645 Lys Val Glu Thr Leu Pro Tyr Val Pro Glu Pro Val Lys Val Ala Ile 1650 1655 1660 Ala Glu Asn Leu Leu Asp Val Ile Lys Asp Thr Arg Ser Lys Glu Ala 1665 1670 1675 1680 Thr Pro Val Ala Ala Gly Glu Ala Gly Asp Glu Asp Gly Ala Val Ile 1685 1690 1695 Val Ser Lys Ala Ala His Ser Ser Arg Leu Thr Asn Ser Thr Pro Lys 1700 1705 1710 Thr Val Lys Glu Pro Arg Ala Glu Thr Val Asn Thr Ser Gln Ser Asp 1715 1720 1725 Asp Met Val Ser Ser Arg Thr Leu Thr Arg Arg Gln His Ala Leu Ser 1730 1735 1740 Leu Asn Val Thr Ser Glu Gln Glu Pro Ser Ala Val Ala Thr Pro Lys 1745 1750 1755 1760 Lys Arg Thr Arg Lys Ile Lys Glu Thr Pro Glu Ser Ser Glu Arg Thr 1765 1770 1775 Cys Ser Asp Leu Lys Val Ala Pro Glu Asn Gln Leu Thr Ala Gln Asn 1780 1785 1790 Pro Pro Ala Pro Arg Arg Arg Lys Lys Lys Asp Val Ser Gln Gly Thr 1795 1800 1805 Leu Pro Ser Ser Gly Ala Val Glu Pro Glu Pro Glu Pro Gln Gly Th r 1810 1815 1820 Pro Gly Arg Leu Arg Leu Arg Thr Gln Pro Pro Glu Pro Ala Ala Glu 1825 1830 1835 1840 Glu Thr Pro Ser Arg Thr Lys Val Arg Leu Ser Ser Val Arg Lys Gly 1845 1850 1855 Thr Pro Arg Arg Leu Lys Lys Ser Val Glu Asn Gly Gln Ser Ile Glu 1860 1865 1870 Ile Leu Asp Asp Leu Lys Gly Ser Glu Ala Ala Ser His Asp Gly Thr 1875 1880 1885 Val Thr Glu Leu Arg Asn Ala Asn Leu Glu Asp Thr Gln Asn Met Glu 1890 1895 1900 Tyr Lys Gln Asp Glu His Ser Asp Gln Gln Pro Pro Leu Lys Arg Lys 1905 1910 1915 1920 Arg Val Arg Glu Arg Glu Val Ser Val Ser Ser Val Thr Glu Glu Pro 1925 1930 1935 Lys Leu Asp Ser Ser Gln Leu Pro Leu Gln Thr Gly Leu Asp Val Pro 1940 1945 1950 Ala Thr Pro Arg Lys Arg Gly Arg Pro Arg Lys Val Val Pro Leu Glu 1955 1960 1965 Ala Asp Gly Gly Thr Thr Thr Gly Lys Glu Gln Thr Ser Pro Gln Lys Lys 1970 1975 1980 Asp Val Pro Val Val Arg Arg Ser Thr Arg Asn Thr Pro Ala Arg Asn 1985 1990 1995 2000 Val Ser Thr Leu Lys Lys Ser Val Leu Val Pro Asn Lys Glu Ala Ala 2005 2010 2015 Leu Val Val Thr Ser Lys Arg Arg Pro Thr Lys Lys Ser Ala Glu Glu 2020 2025 2030 Ser Ser Lys Asp Pro Ser Ala Ala Val Ser Asp Trp Ala Gly Gly Ala 2035 2040 2045 Ala His Thr Glu Ser Ala Asp Arg Arg Asp Gly Leu Leu Ala Ala Ala 2050 2055 2060 Ala Leu Thr Pro Ser Ala Gln Gly Thr Arg Thr Arg Ser Arg Arg Thr 2065 2070 2075 2080 Met Leu Leu Thr Asp Ile Ser Glu Pro Lys Thr Glu Pro Leu Phe Pro 2085 2090 2095 Pro Pro Ser Val Lys Val Pro Lys Lys Lys Ser Lys Ala Glu Asn Met 2100 2105 2110 Glu Ala Ala Ala Gln Leu Lys Glu Leu Val Ser Asp Leu Ser Ser Gln 2115 2120 2125 Phe Val Val Ser Pro Pro Ala Leu Arg Thr Arg Gln Lys Ser Ile Ser 2130 2135 2140 Asn Thr Ser Lys Leu Leu Gly Glu Leu Glu Ser Asp Pro Lys Pro Leu 2145 2150 2155 2160 Glu Ile Ile Glu Gln Lys Pro Lys Arg Ser Arg Thr Val Lys Thr Arg 2165 2170 2175 Ala Ser Arg Asn Thr Gly Lys Gly Ser Ser Trp Ser Pro Pro Pro Val 2180 2185 2190 Glu Ile Lys Leu Val Ser Pro Leu Ala Ser Pro Val Asp Glu Ile Lys 2195 2200 2205 Thr Gly Lys Pro Arg Lys Thr Ala Glu Il e Ala Gly Lys Thr Leu Gly 2210 2215 2220 Arg Gly Arg Lys Lys Pro Ser Ser Phe Pro Lys Gln Ile Leu Arg Arg 2225 2230 2235 2240 Lys Met Leu <210> 13 <211> 7215 <212> DNA <213> Homo sapiens <220> <221> CDS <222> (125) .. (6922) <400> 13 tgcggctcga gggggccagc gctgacggtg gcgggtacgg caggctcgcg ggcgccgggc 60 ttcgttacat aatctcggac cggaggagcg gtggcacatg gcggcggaac ggcgctgtgg 120 aagt atg cga gac tta aga gct caa gtg act agt ggt ctc ctg cca ttt 169 Met Arg Asp Leu Arg Ala Gln Val Thr Ser Gly Leu Leu Pro Phe 1 5 10 15 cca gaa gtg act ctt caa gcc ctt gga gaa gac gaa ata aca tta gaa 217 Pro Glu Val Thr Leu Gln Ala Leu Gly Glu Asp Glu Ile Thr Leu Glu 20 25 30 tct gtg ctt cgt gga aag ttt gct gcg ggg aaa aat gga ctt gct tgc 265 Ser Val Leu Arg Gly Lys Phe Ala Ala Gly Lys Asn Gly Leu Ala Cys 35 40 45 ttg gct tgt ggt cca caa ctt gag gta gta aac tct ata aca gga gag 313 Leu Ala Cys Gly Pro Gln Leu Glu Val Val Asn Ser Ile Thr Gly Glu 50 55 60 cga ttg tct gct tac aga ttc agt gga gtc aat gaa cag cct cct gta 361 Arg Leu Ser Ala Tyr Arg Phe Ser Gly Val Asn Glu Gln Pro Pro Val 65 70 75 gtt tta gct gtg aaa gaa ttc tct tgg cag aag aga act gga tta tta 409 Val Leu Ala Val Lys Glu Phe Ser Trp Gln Lys Arg Thr Gly Leu Leu 80 85 90 9 5 ata gga ttg gaa gaa aca gaa ggg agt gtt ctc tgt ctt tat gac ctt 457 Ile Gly Leu Glu Glu Thr Glu Gly Ser Val Leu Cys Leu Tyr Asp Leu 100 105 110 gga ata tca aaa gta gtt aaa gca gtt gtt ctt cct g agg gta aca 505 Gly Ile Ser Lys Val Val Lys Ala Val Val Leu Pro Gly Arg Val Thr 115 120 125 gct att gaa cct ata att aat cat gga gga gcc agt gca agc act cag 553 Ala Ile Glu Pro Ile Ile Asn His Gly Gly Ala Ser Ala Ser Thr Gln 130 135 140 cat tta cat cca agt ctg cga tgg ctt ttt gga gtg gca gct gtg gtc 601 His Leu His Pro Ser Leu Arg Trp Leu Phe Gly Val Ala Ala Val Val 145 150 155 act gat gtt gga cag atc ctt ctt att gac cta tgt ttg gat gac ttg 649 Thr Asp Val Gly Gln Ile Leu Leu Ile Asp Leu Cys Leu Asp Asp Leu 160 165 170 175 tca tgc aat caa aat gaa gtt gaa gca tca gat ctt gaa gtt cta act Cys Asn Gln Asn Glu Val Glu Ala Ser Asp Leu Glu Val Leu Thr 180 185 190 ggt atc cca gct gaa gta cca cac att aga gaa agt gtg atg aga gaa 745 Gly Ile Pro Ala Glu Val Pro His Ile Arg Glu Ser Val Met Arg Glu 195 200 205 ggg cgc cat ctg tgt ttc cag tta gta agt cca aca gga aca gcc gtt 793 Gly Arg His Leu Cys Phe Gln Leu Val Ser Pro Thr Gly Thr Ala Val 210 215 220 tca act ctt agt tac ata agc agg aca aat cag ctt gct gca ggt ttt 841 Ser Thr Leu Ser Tyr Ile Ser Arg Thr Asn Gln Leu Ala Ala Gly Phe 225 230 235 tct gat ggc tat cta gca ctt tgg aac atg aaa agc atg aaa aga gaa 889 Ser Asp Gly Tyr Leu Ala Leu Trp Asn Met Lys Ser Met Lys Arg Glu 240 245 250 255 tat tac ata caa ttg gaa agt gga caa gtt cct gta tat gct gtc act 937 Tyr Tyr Ile Gln Leu Glu Ser Gly Gln Val Pro Val Tyr Ala Val Thr 260 265 270 270 ttt caa gaa cct gag aat gat cgt cgg aat tgc tgc tac ttg tgg gct 985 Phe Gln Glu Pro Glu Asn Asp Arg Arg Asn Cys Cys Tyr Leu Trp Ala 275 280 285 gtt cag tct aca caa gat agt gat ggg gat gtt cat ctg 1033 Val Gln Ser Thr Gln Asp Ser Glu Gly Asp Val Leu Ser Leu His Leu 290 295 300 ctg cag ctg gcc ttt ggt aat aga aag tgt ttg gca tca gga caa atc 1081 Leu Gln Leu Ala Phe Gly Asn Arg Lys Cys Leu Ala Ser Gly Gln Ile 305 310 315 tta tat gag ggg tta gaa tac tgt gaa gaa aga tac acc ctg gac ctg 1129 Leu Tyr Glu Gly Leu Glu Tyr Cys Glu Glu Arg Tyr Thr Leu Asp Leu 320 325 330 335 acagt atg ttc cct ttg agg gga cag acg agt aat acc aaa ttg 1177 Thr Gly Gly Met Phe Pro Leu Arg Gly Gln Thr Ser Asn Thr Lys Leu 340 345 350 ttg gga tgc cag agt ata gag aaa ttt cga tct cat ggt gac agg gag12 Leu Gly Cys Gln Ser Ile Glu Lys Phe Arg Ser His Gly Asp Arg Glu 355 360 365 gaa ggc gtg aat gaa gct cta tcg cct gac act agt gtt tca gtc ttt 1273 Glu Gly Val Asn Glu Ala Leu Ser Pro Asp Thr Ser Val Ser Val Phe 370 375 380 acc tgg cag gtg aat ata tat gga cag gga aag cct tct gta tat ttg 1321 Thr Trp Gln Val Asn Ile Tyr Gly Gln Gly Lys Pro Ser Val Tyr Leu 385 390 395 395 ggg ctt ttt gat ata aat cgt tgg tat cat gca caa atg cca gat tcg 1369 Gly Leu Phe Asp Ile Asn Arg Trp Tyr His Ala Gln Met Pro Asp Ser 400 405 410 415 tta agg tca gga gaa tat cta cat aat tgc tct tat ttt gca ctg tgg 1417 Leu Arg Ser Gly Glu Tyr Leu His Asn Cys Ser Tyr Phe Ala Leu Trp 420 425 430 tca ttg gag tct gtt gta agt agg act tct cca cat ggc atc ttg gat 1465 Ser Leu Glu Ser Val Val Ser Arg Thr Ser Pro His Gly Ile Leu Asp 435 440 445 ata tta gta cat gag aga agt tta aat aga gga gtc cct cct tca tat 1513 Ile Leu Val His Glu Arg Ser Leu Asn Arg Gly Val Pro Pro Ser Tyr 450 455 460 cca cct ccc gag cag ttt ttt aat cca agc act tat aat ttt gat gcc 1561 Pro Pro Pro Glu Gln Phe Phe Asn Pro Ser Thr Tyr Asn Phe Asp Ala 465 470 475 act tgt ttg tta aac tcg gga gtt gtt cat tta act tgt act ggc ttt 1609 Thr Cys Leu Leu Asu Ser Gly Val Val His Leu Thr Cys Thr Gly Phe 480 485 490 495 cag aag gag act ttg act ttt tta aag aaa tca ggt cca tca ctc aat 1657 Gln Lys Glu Thr Leu Thr Phe Leu Lys Lys Ser Gly Pro Ser Leu Asn 500 505 510 gaa ctc att cct gat ggt tat aat cga tgt ctt gta gct ggc ctt ctt 1705 Glu Leu Ile Pro Asp Gly Tyr Asn Arg Cys Leu Val Ala Gly Leu Leu 515 520 525 tcc cca aga ttt gtt gat gtt cag cct tcc agt tta agc caa gaa gaa 1753 Ser Pro Arg Phe Val Asp Val Gln Pro Ser Ser Leu Ser Gln Glu Glu 530 535 540 cag tta gaa gct ata ttg tca gca gca att cag act agt tcc ctg gga 1801 Gln Leu Glu Ala Ile Leu Ser Ala Ala Ile Gln Thr Ser Ser Leu Gly 545 550 555 ctt ttg act ggt tat atc cga aga tgg ata aca gaa gaa caa cca aat 1849 Leu Leu Thr Gly Tyr Ile Arg Arg Trp Ile Thr Glu Glu Gln Pro Asn 560 565 570 570 575 tct gcc act aat ttg cgc ttt gtt ctt gaa tgg acg tgg aat aaa gtg 1897 Ser Ala Thr Asn Leu Arg Phe Val Leu Glu Trp Thr Trp Asn Lys Val 580 585 590 gtt ctc aca aaa gag gaa ttt gac aga cta tgt gtg cca tta ttt Val Leu Thr Lys Glu Glu Phe Asp Arg Leu Cys Val Pro Leu Phe Asp 595 600 605 ggt tcg tgt cat ttc atg gat cca caa act ata cag tct atc cag caa 1993 Gly Ser Cys His Phe Met Asp Pro Gln Thr Ile Gln Ser Ile Gln Gln 610 615 620 tgc tat ttg ctt ctt agc aat ctt aat ata gtc ttg agc tgt ttt gca 2041 Cys Tyr Leu Leu Leu Ser Asn Leu Asn Ile Val Leu Ser Cys Phe Ala 625 630 630 635 tca gaa gcc cga gat c gct gag aga gga ctg ata gac tta agc aat 2089 Ser Glu Ala Arg Glu Ile Ala Glu Arg Gly Leu Ile Asp Leu Ser Asn 640 645 650 655 aag ttt gtg gtt tcc cac ctc atc tgt cag tat gca caa gtg gtt ctt Phe Val Val Ser His Leu Ile Cys Gln Tyr Ala Gln Val Val Leu 660 665 670 tgg ttc tct cat tct ggg ctt tta cca gaa ggc ata gat gat tct gtg 2185 Trp Phe Ser His Ser Gly Leu Leu Pro Glu Gly Ile Asp Asp Ser Val 675 680 685 cag ttg tca agg tta tgc tac aac tac cct gta att cag aac tac tac 2233 Gln Leu Ser Arg Leu Cys Tyr Asn Tyr Pro Val Ile Gln Asn Tyr Tyr 690 695 700 acc agt cgt cga cag aag ttt gag cgt tta tca aga ggg aag tgg aat 2281 Thr Ser Arg Arg Gln Lys Phe Glu Arg Leu Ser Arg Gly Lys Trp Asn 705 710 715 ccc gat tgc ttg atg att gat gga ctg gtt tct cag tta gga gag cga 2329 Pro Asp Cys Leu Met Ile Gly Leu Val Ser Gln Leu Gly Glu Arg 720 725 730 735 att gag aag ttg tgg aaa cga gat gaa gga ggc aca gga aaa tat cct 2377 Ile Glu Lys Leu Trp Lys Arg Asp Glu Gly Gly Thr Gly Lys Tyr Pro 740 745 750 cct gct agt ctg cat gca gta ctt gat atg tac cta tta gac ggc gtt 2425 Pro Ala Ser Leu His Ala Val Leu Asp Met Tyr Leu Leu Asp Gly Val 755 760 765 act gaa gca gcc aaa cac tct att acc att tat ttg cta ctt gat att 2473 Thr Glu Ala Ala Lys His Ser Ile Thr Ile Tyr Leu Leu Leu Asp Ile 770 775 780 atg tat tcc ttt ccc aac aaa aca gac act ccc att gaa tct ttc cca 2521 Met Tyr Ser Phe Pro Asn Lys Thr Asp Thr Pro Ile Glu Ser Phe Pro 785 790 795 act gta ttt gcc att tct tgg ggc caa gtt aaa ctt att cag ggg ttt 2569 Thr Val Phe Ala Ile Ser Trp Gly Gln Val Lys Leu Ile Gln Gly Phe 800 805 810 815 815 tgg ttg ata gat cat aat gac tat gag agt ggt ttg gat ctt ttg ttt 2617 Trp Leu Ile Asp His Asn Asp Tyr Glu Ser Gly Leu Asp Leu Leu Phe 820 825 830 cat cca gct act gca aaa cct ttg tca tgg caa cat tca aag att att 2665 His Pro Ala Thr Ala Lys Pro Leu Ser Trp Gln His Ser Lys Ile Ile 835 840 845 cag gca ttc atg agt cag ggc gag cac aga caa gcc ctc aga tat att 2713 Gln Ala Phe Met Ser Gln Gly Glu His Arg G ln Ala Leu Arg Tyr Ile 850 855 860 cag aca atg aag cca aca gtg tcc agt ggt aac gat gtt atc ctt cac 2761 Gln Thr Met Lys Pro Thr Val Ser Ser Gly Asn Asp Val Ile Leu His 865 870 875 ctc act gtt ttg ctt ttt aat agg tgt atg gtt gaa gcc tgg aat ttt 2809 Leu Thr Val Val Leu Leu Phe Asn Arg Cys Met Val Glu Ala Trp Asn Phe 880 885 890 895 ttg cgg caa cat tgc aat agg ttg aat ata gag gag tta 857 857 Arg Gln His Cys Asn Arg Leu Asn Ile Glu Glu Leu Leu Lys His 900 905 910 atg tat gaa gtc tgt cag gaa atg ggc ttg atg gaa gat tta ctg aag 2905 Met Tyr Glu Val Cys Gln Glu Met Gly Leu Met Glu Lys 915 920 925 tta cca ttt aca gac act gag cag gaa tgt tta gtg aaa ttt ttg cag 2953 Leu Pro Phe Thr Asp Thr Glu Gln Glu Cys Leu Val Lys Phe Leu Gln 930 935 940 tcc agt gcc agc gtt cag aat catgaa ctt tta gtg cac cat ttg 3001 Ser Ser Ala Ser Val Gln Asn His Glu Phe Leu Leu Val His His Leu 945 950 955 cag cgt gcc aat tat gtg cct gcc ttg aag ctg aac caa act ctg aag 3049 Gln Arg Ala Asn Tyr Val Pro Ala Leu Lys Leu Asn Gln Thr Leu Lys 960 965 970 975 att aat gtt atg aat gat cgt gat cct cgt ttg cgg gag aga tca ctg 3097 Ile Asn Val Met Asn Asp Arg Asp Pro Arg Leu Arg Glu Arg Ser Leu 980 985 990 gct cga aat tct ata tta gac cag tat gga aaa atc ctt cct aga gtc 3145 Ala Arg Asn Ser Ile Leu Asp Gln Tyr Gly Lys Ile Leu Pro Arg Val 995 1000 1005 cat cga aaa tta gcc att gaa cga gct aag cct tat cat ctg tca aca 3193 His Arg Lys Leu Ala Ile Glu Arg Ala Lys Pro Tyr His Leu Ser Thr 1010 1015 1020 tca tca gtt ttt cga tta gtt tct aga ccc aaa cca tta tca gca gtt 3241 Ser Ser Val Phe Arg Leu Val Ser Arg Pro Lys Pro Leu Ser Ala Val 1025 1030 1035 cca aag caa gtt gta aca gga act gtg ttg aca aga tct gtt ttc atc 3289 Pro Lys Gln Val Val Thr Gly Thr Val Leu Thr Arg Ser Val Phe Ile 1040 1045 1050 1055 aac aat gtg tta tct aaa att gga gaa gtt tgg gca agc aaa gaa cct 3337 Asn Asn Val Leu Ser Lys Ile Gly Glu Val Trp Ala Ser Lys Glu Pro 1060 1065 1070 ata aat agc acc aca cct ttc aat agt tct aaa ata gaa gaa cca tct 3385 Ile Asn Ser Thr Thr Pro Phe Asn Ser Ser Lys Ile Glu Glu Pro Ser 1075 1080 1085 cct ata gtg tat tcg ctc cca gct cca gag ctg cct gag gca ttt ttt 3433 Pro Ile Val Tyr Ser Leu Pro Ala Pro Glu Leu Pro Glu Ala Phe Phe 1090 1095 1100 gga aca cca att tca aaa gca tca caa aaa att tct aga ctg cta gat 3481 Gly Thr Pro Ile Ser Lys Ala Ser Gln Lys Ile Ser Arg Leu Leu Asp 1105 1110 1115 ttg gtt gtt cag cct gtc ccc cgg cct tct cag tgt tcg gag ttt att 3529 Leu Val Val Gln Pro Val Pro Arg Pro Ser Gln Cys Ser Glu Phe Ile 1120 1125 1130 1135 cag caa agc tcc atg aaa tct cct ttg tac cta gta tcc cgt tca ctg 3577 Gln Gln Ser Ser Met Lys Ser Pro Leu Tyr Leu Val Ser Arg Ser Leu 1140 1145 1150 ccc tca agt tcg caa tta aaa gga tcg cct cag gcc atc tcc agg gct 3625 Pro Ser Ser Ser Ser Ser Gln Leu Lys Gly Ser Pro Gln Ala Ile Ser Arg Ala 1155 1160 1165 tca gaa tta cat ttg ctt gaa act cct ctt gta gtt aag aaa gct aaa 3673 Ser Glu Leu His Leu Leu Glu Thr Pro Leu Val Val Lys Lys Ala Lys 1170 117 5 1180 agt ttg gcc atg tca gtt act act tct gga ttt tct gag ttc act cct 3721 Ser Leu Ala Met Ser Val Thr Thr Ser Gly Phe Ser Glu Phe Thr Pro 1185 1190 1195 cag tcc atc ctg agg tct act cct cga tca aca cct tta gca tct ccc 3769 Gln Ser Ile Leu Arg Ser Thr Pro Arg Ser Thr Pro Leu Ala Ser Pro 1200 1205 1210 1215 tct cca tca cct gga agg tct cct caa cga ctt aaa gaa act aga att 3817 Ser Pro Ser Pro Gly Arg Ser Pro Gln Arg Leu Lys Glu Thr Arg Ile 1220 1225 1230 tca ttt gtg gaa gaa gat gtc cac cca aaa tgg att cct ggg gct gca 3865 Ser Phe Val Glu Glu Asp Val His Pro Lys Trp Ile Pro Gly Ala Ala 1235 1240 1245 gat gatg aaa tta gaa gta ttt act aca cct aaa aaa tgt gca gtt 3913 Asp Asp Ser Lys Leu Glu Val Phe Thr Thr Pro Lys Lys Cys Ala Val 1250 1255 1260 cca gtg gaa act gaa tgg ccg aag agc aaa gat agg acc aca tct ttt 3961 Pro Val Glu Thr Glu Trp Pro Lys Ser Lys Asp Arg Thr Thr Ser Phe 1265 1270 1275 ttc ctg aac agc cct gaa aag gag cat caa gaa atg gat gag ggg tca 4009 Phe Leu Asn Ser Pro Glu Lys Glu His Gln Glu Met Asp Glu Gly Ser 1280 1285 1290 1295 caa agt tta gag aaa ctg gat gtg agc aaa gga aac agc agt gtt tca 4057 Gln Ser Leu Glu Lys Leu Asp Val Ser Lys Gly Asn Ser Ser Val Ser 1300 1305 1310 atc aca tcc gat gag act acc tta gag tat cag gat gca ccg tca ccg 4105 Ile Thr Ser Asp Glu Thr Thr Leu Glu Tyr Gln Asp Ala Pro Ser Pro 1315 1320 1325 gaa gac ctt gaa gag act gtt ttc acg gcc tct aag ccc aaa agc tct 4153 Glu Asp Leu Glu Glu Thr Val Phe Thr Ala Ser Lys Pro Lys Ser Ser 1330 1335 1340 tcc act gca cta act act aat gta act gaa caa act gaa aag gat gga 4201 Ser Thr Ala Leu Thr Thr Asn Val Thr Glu Gln Thr Glu Lys Asp Gly 1345 1350 1355 gat aaa gat gta ttt gca tca gaa gta act cct tca gac cta cag aaa 4249 Asp Lys Asp Val Phe Ala Ser Glu Val Thr Pro Ser Asp Leu Gln Lys 1360 1365 1370 1375 caa atg ggc aat tta gaa gat gca gaa aca aag gat ctc tta gtt gca 4297 Gln Met Gly Asn Leu Glu Asp Ala Glu Thr Lys Asp Leu Leu Val Ala 1380 1385 1390 gca gag gca ttt tca gaa ttg aat cac tt a agc ccg gtt caa gga act 4345 Ala Glu Ala Phe Ser Glu Leu Asn His Leu Ser Pro Val Gln Gly Thr 1395 1400 1405 gaa gct tct ctt tgt gca cca tca gtc tat gaa ggg aaa atc ttc acc 4393 Glu Ala Ser Leu Cys Ala Pro Ser Val Tyr Glu Gly Lys Ile Phe Thr 1410 1415 1420 cag aag tcc aag gta cca gtg ttg gac gaa gga tta aca tct gtt gaa 4441 Gln Lys Ser Lys Val Pro Val Leu Asp Glu Gly Leu Thr Ser Val Glu 1425 1430 1435 acc tac acc cct gca att aga gca aat gac aat aaa tct atg gct gat 4489 Thr Tyr Thr Pro Ala Ile Arg Ala Asn Asp Asn Lys Ser Met Ala Asp 1440 1445 1450 1455 gtc ctt ggt gat ggt gga aac tcc tcg ctc act atc tctga ggt cct 4537 Val Leu Gly Asp Gly Gly Asn Ser Ser Leu Thr Ile Ser Glu Gly Pro 1460 1465 1470 att gtc tct gag cgc agg ctt aac cag gaa gta gcg ctg aac tta aaa 4585 Ile Val Ser Glu Arg Arg Leu Asn Gln Glu Val Ala Leu Asn Leu Lys 1475 1480 1485 gaa gat cat gaa gta gaa gtt ggt gta cta aaa gaa agt gtt gac tta 4633 Glu Asp His Glu Val Glu Val Gly Val Leu Lys Glu Ser Val Asp Leu 1490 1 495 1500 cca gaa gaa aag ctt cca att tct gac agc cct cct gat act caa gaa 4681 Pro Glu Glu Lys Leu Pro Ile Ser Asp Ser Pro Pro Asp Thr Gln Glu 1505 1510 1515 att cat gtg att gaa caa gaa aag ctt gaa gca gat tca gga gaa 4729 Ile His Val Ile Glu Gln Glu Lys Leu Glu Ala Gln Asp Ser Gly Glu 1520 1525 1530 1535 gag gct agg aat ctt tca ttt aat gag tta tat ccc tct gga aca ctt 4777 Glu Ala Arg Asn Leu Ser Phe Asn Glu Leu Tyr Pro Ser Gly Thr Leu 1540 1545 1550 aag ctt cag tac aat ttt gat act att gac caa cag ttt tgt gac tta 4825 Lys Leu Gln Tyr Asn Phe Asp Thrle Ile Asp Gln Gln Phe Cys Asp Leu 1555 1560 1565 gct gat aac aaa gac act gct gaa tgt gac att gct gaa gta gat ggg 4873 Ala Asp Asn Lys Asp Thr Ala Glu Cys Asp Ile Ala Glu Val Asp Gly 1570 1575 1580 gaa ctt ttt gtg gct caa agc aac ttt acc ttg ata gtgga 921 Glu Leu Phe Val Ala Gln Ser Asn Phe Thr Leu Ile Leu Glu Gly Glu 1585 1590 1595 gaa gga gaa gtt gag cca ggt gat ttt gca tca tct gat gtg tta cct 4969 Glu Gly Glu Val Glu Pro Gly Asp Phe Ala Ser Ser Asp Val Leu Pro 1600 1605 1610 1615 aaa gca gct aac aca gca act gaa gaa aaa ctt gta tgc agt ggg gaa 5017 Lys Ala Ala Asn Thr Ala Thr Glu Glu Lys Leu Val Cys Ser Gly Glu 1620 1625 1630 aat gat aat cat gga caa att gca aat ttg cca tct gcc gta act agt 5065 Asn Asp Asn His Gly Gln Ile Ala Asn Leu Pro Ser Ala Val Thr Ser 1635 1640 1645 gac caa aag tcc caa aaa gta gac act tta cca tat gtg cct gaa cct 5113 Asp Gln Lys Ser Gln Lys Val Asp Thr Leu Pro Tyr Val Pro Glu Pro 1650 1655 1660 att aaa gta gca att gca gaa aat tta cta gat gta att aaa gac aca 5161 Ile Lys Val Ala Ile Ala Glu Asn Leu Leu Asp Val Ile Lys Asp Thr 1665 1670 1675 aga agt aaa gaa att act tca gat aca atg gaa cag tcc att cat gaa 5209 Arg Ser Lys Glu Ile Thr Ser Asp Thr Met Glu Gln Ser Ile His Glu 1680 1685 1690 1695 aca ata cct tta gtg agc caa aac ata atg tgt ccc act aaa ttg gtc 5257 Thr Ile Pro Leu Val Ser Gln Asn Ile Met Cys Pro Thr Lys Leu Val 1700 1705 1710 aaa tct gca ttt aag act gct cag gaa aca agc aca atg act atg aat 5305 Lys Ser Ala Phe Lys Thr Ala Gln Glu Thr Ser Thr Met Thr Met Asn 1715 1720 1725 gtc agc cag gtt gat gac gtg gtt tcc tcc aaa act cgt acg aga ggt 5353 Val Ser Gln Val Asp Asp Val Val Ser Ser Lys Thr Arg Thr Arg Gly 1730 1735 1740 caa cgt atc caa aac gtg aat gtc aaa tca gca caa cag gaa gca tca 5401 Gln Arg Ile Gln Asn Val Asn Val Lys Ser Ala Gln Gln Glu Ala Ser 1745 1750 1755 gca gat gtt gct act cct aag atg cca ggg cag tca gtc agg aag aaa 5449 Ala Asp Val Ala Thr Pro Lys Met Pro Gly Gln Ser Val Arg Lys Lys 1760 1765 1770 1775 act agg aag gca aaa gaa att tct gaa gct tct gaa aac atc tat tct 5497 Thr Arg Lys Ala Lys Glu Ile Ser Glu Ala Ser Glu Asn Ile Tyr Ser 1780 1785 1790 gat gtc aga gga cta ttt cag aac cag caa ata cct caa aat tct gtt 5545 Asp Val Arg Gly Leu Phe Gln Asn Gln Gln Ile Pro Gln Asn Ser Val 1795 1800 1805 acg cct agg aga gga agg aga aag aaa gaa gtt aat cag gac ata cta 5593 Thr Pro Arg Arg Gly Arg Arg Lys Lys Glu Val Asn Gln Asp Ile Leu 1810 1 815 1820 gaa aac acc agt tct gtg gaa caa gaa tta cag atc act aca ggt agg 5641 Glu Asn Thr Ser Ser Val Glu Gln Glu Leu Gln Ile Thr Thr Gly Arg 1825 1830 1835 gaa tca aaa aga tta aaa tca tct cag ctg ttg cca gca gtt gaa 5689 Glu Ser Lys Arg Leu Lys Ser Ser Gln Leu Leu Glu Pro Ala Val Glu 1840 1845 1850 1855 gaa act act aaa aaa gaa gtt aag gtt tca tct gtt aca aaa agg act 5737 Glu Thr Thr Lys Lys Glu Val Lys Val Ser Ser Val Thr Lys Arg Thr 1860 1865 1870 cct aga aga att aaa aga tct gta gaa aat cag gaa agt gtt gaa att 5785 Pro Arg Arg Ile Lys Arg Ser Val Glu Asn Gln Glu Ser Val Glu Ile 1875 1880 1885 ata aat gat cta aaa gtt agt acg gta aca agt cct agc aga atg atc 5833 Ile Asn Asp Leu Lys Val Ser Thr Val Thr Ser Pro Ser Arg Met Ile 1890 1895 1900 aga aaa ttg aga agt act aat tta gat gct tct gaa aat aca gga aat 5881 Arg Lys Leu Arg Ser Thr Asn Leu Asp Ala Ser Glu Asn Thr Gly Asn 1905 1910 1915 aag caa gat gat aaa tcc agt gac aag cag ctg cgt att aaa cat gtt 5929 Lys Gln Asp Asp Lys Se r Ser Asp Lys Gln Leu Arg Ile Lys His Val 1920 1925 1930 1935 aga agg gtc aga ggg aga gaa gtt agt cca tca gat gtg aga gaa gac 5977 Arg Arg Val Arg Gly Arg Glu Val Ser Pro Ser Asp Val Arg Glu Asp 1940 1945 1950 tcc aac ctt gag tca tct cag ttg act gtt caa gca gaa ttt gat atg 6025 Ser Asn Leu Glu Ser Ser Gln Leu Thr Val Gln Ala Glu Phe Asp Met 1955 1960 1965 tct gcc ata cct aga aaa cgt ggt aga cca aga atc aat cca tct 6073 Ser Ala Ile Pro Arg Lys Arg Gly Arg Pro Arg Lys Ile Asn Pro Ser 1970 1975 1980 gaa gat gta gga tct aag gct gtt aag gaa gag aga agc ccc aag aag 6121 Glu Asp Val Gly Ser Lys Ala Val Lys Glu Glu Arg Ser Pro Lys Lys 1985 1990 1995 aaa gaa gct ccc agc att aga agg aga tct aca aga aat acc cca gct 6169 Lys Glu Ala Pro Ser Ile Arg Arg Arg Ser Thr Arg Asn Thr Pro Ala 2000 2005 2010 2015 aaa agt gaa aat gtt gat gtt gga aaa cca gct tta gga aaa tcc att 6217 Lys Ser Glu Asn Val Asp Val Gly Lys Pro Ala Leu Gly Lys Ser Ile 2020 2025 2030 tta gtg cca aac gag gaa ctt tcg atg gtg atg agc tct aag aaa aaa 6265 Leu Val Pro Asn Glu Glu Leu Ser Met Val Met Ser Ser Lys Lys Lys 2035 2040 2045 ctt aca aaa aag act gaa agt caa agc caa aaa cgt tca ttg cac tca 6313 Leu Thr Lys Lys Thr Glu Ser Gln Ser Gln Lys Arg Ser Leu His Ser 2050 2055 2060 gta tca gaa gaa cgc aca gat gaa atg aca cat aaa gaa aca aat gag 6361 Val Ser Glu Glu Glu Arg Thr Asp Glu Met Thr His Lys Glu Thr Asn Glu 2065 2070 2075 cag gaa gaa aga ttg ctc gcc aca gct tcc ttc act aaa tca tcc cgc 6409 Gln Glu Glu Arg Leu Leu Ala Thr Ala Ser Phe Thr Lys Ser Ser Arg 2080 2085 2090 2095 agc agc agg act cgg tct agc aag gcc atc ttg gttg ttg cc tct 6457 Ser Ser Arg Thr Arg Ser Ser Lys Ala Ile Leu Leu Pro Asp Leu Ser 2100 2105 2110 gaa cca aac aat gag cct tta ttt tct cca gcg tca gaa gtt cca agg 6505 Glu Pro Asn Asn Glu Pro Leu Phe Ser Pro Ala Ser Glu Val Pro Arg 2115 2120 2125 aaa gca aaa gct aaa aaa ata gag gtt cct gca cag ctg aaa gaa tta 6553 Lys Ala Lys Ala Lys Lys Ile Glu Val Pro Ala Gln Leu Lys Glu Leu 2130 21 35 2140 gtt tcg gat tta tct tct cag ttt gtc atc tca cct cct gct tta agg 6601 Val Ser Asp Leu Ser Ser Gln Phe Val Ile Ser Pro Pro Ala Leu Arg 2145 2150 2155 agc aga caa aaa aac aca tcc aat aag aac aag ctt gaa gat gaa ctg 6649 Ser Arg Gln Lys Asn Thr Ser Asn Lys Asn Lys Leu Glu Asp Glu Leu 2160 2165 2170 2175 aaa gat gat gca caa tca gta gaa act ctg gga aag cca aaa gcg aaa 6697 Lys Asp Asp Alu Gln Ser Val Thr Leu Gly Lys Pro Lys Ala Lys 2180 2185 2190 cga atc agg acg tca aaa aca aaa caa gca agc aaa aac aca gaa aaa 6745 Arg Ile Arg Thr Ser Lys Thr Lys Gln Ala Ser Lys Asn Thr Glu Lys 2195 2200 2205 gaa agt gct tgg tca ctt cct ccc ata gaa att cgg ctg att tcc ccc 6793 Glu Ser Ala Trp Ser Leu Pro Pro Ile Glu Ile Arg Leu Ile Ser Pro 2210 2215 2220 ttg gct agc cca gct gac gga gtc aag agc aaa cca aga act aca 68 Leu Ala Ser Pro Ala Asp Gly Val Lys Ser Lys Pro Arg Lys Thr Thr 2225 2230 2235 gaa gtg aca gga aca ggt ctt gga agg aac aga aag aaa ctg tct tcc 6889 Glu Val Thr Gly Thr Gl y Leu Gly Arg Asn Arg Lys Lys Leu Ser Ser 2240 2245 2250 2255 tat cca aag caa att tta cgc aga aaa atg ctg taatttcttg ggaagatttt 6942 Tyr Pro Lys Gln Ile Leu Arg Arg Lys Mata Leu 2260 2265 aatgtacgt gag tatt gat tatt gat tatt gat tagtatt gat gag ttatataaat tattgtaaat ttttatgtaa acagaaggtc ttcaataagt 7062 aaagtaactc catatggagt gattgtttca gtccaggcaa tttttcttt ttatattaag 7122 acttcataca tttatatatg taaatatggc ttattaatgg aatgttaaat aaaatgtata 7aaaaaactaa <210> 14 <211> 2266 <212> PRT <213> Homo sapiens <400> 14 Met Arg Asp Leu Arg Ala Gln Val Thr Ser Gly Leu Leu Pro Phe Pro 1 5 10 15 Glu Val Thr Leu Gln Ala Leu Gly Glu Asp Glu Ile Thr Leu Glu Ser 20 25 30 Val Leu Arg Gly Lys Phe Ala Ala Gly Lys Asn Gly Leu Ala Cys Leu 35 40 45 Ala Cys Gly Pro Gln Leu Glu Val Val Asn Ser Ile Thr Gly Glu Arg 50 55 60 Leu Ser Ala Tyr Arg Phe Ser Gly Val Asn Glu Gln Pro Pro Val Val 65 70 75 80 Leu Ala Val Lys Glu Phe Ser Trp Gln Lys Arg Thr Gly Leu Leu Ile 85 90 95 Gly Leu Glu Glu Thr Glu Gly Ser Val Leu Cys Leu Tyr Asp Leu Gly 100 105 110 Ile Ser Lys Val Val Lys Ala Val Val Leu Pro Gly Arg Val Thr Ala 115 120 125 Ile Glu Pro Ile Ile Asn His Gly Gly Ala Ser Ala Ser Thr Gln His 130 135 140 Leu His Pro Ser Leu Arg Trp Leu Phe Gly Val Ala Ala Val Val Thr 145 150 155 160 Asp Val Gly Gln Ile Leu Leu Ile Asp Leu Cys Leu Asp Asp Leu Ser 165 170 175 Cys Asn Gln Asn Glu Val Glu Ala Ser Asp Leu Glu Val Leu Thr Gly 180 185 190 Ile Pro Ala Glu Val Pro His Ile Arg Glu Ser Val Met Arg Glu Gly 195 200 205 Arg His Leu Cys Phe Gln Leu Val Ser Pro Thr Gly Thr Ala Val Ser 210 215 220 Thr Leu Ser Tyr Ile Ser Arg Thr Asn Gln Leu Ala Ala Gly Phe Ser 225 230 235 240 Asp Gly Tyr Leu Ala Leu Trp Asn Met Lys Ser Met Lys Arg Glu Tyr 245 250 255 Tyr Ile Gln Leu Glu Ser Gly Gln Val Pro Val Tyr Ala Val Thr Phe 260 265 270 Gln Glu Pro Glu Asn Asp Arg Arg Asn Cys Cys Tyr Leu Trp Ala Val 275 280 285 Gln Ser Thr Gln Asp Ser Glu Gly Asp Val Leu Ser Leu His Leu Leu 290 295 300 Gln Leu Ala Phe Gly Asn Arg Lys Cys Leu Ala Ser Gly Gln Ile Leu 305 310 315 320 Tyr Glu Gly Leu Glu Tyr Cys Glu Glu Arg Tyr Thr Leu Asp Leu Thr 325 330 335 Gly Gly Met Phe Pro Leu Arg Gly Gln Thr Ser Asn Thr Lys Leu Leu 340 345 350 Gly Cys Gln Ser Ile Glu Lys Phe Arg Ser His Gly Asp Arg Glu Glu 355 360 365 Gly Val Asn Glu Ala Leu Ser Pro Asp Thr Ser Val Ser Val Phe Thr 370 375 380 Trp Gln Val Asn Ile Tyr Gly Gln Gly Lys Pro Ser Val Tyr Leu Gly 385 390 395 400 Leu Phe Asp Ile Asn Arg Trp Tyr His Ala Gln Met Pro Asp Ser Leu 405 410 415 Arg Ser Gly Glu Tyr Leu His Asn Cys Ser Tyr Phe Ala Leu Trp Ser 420 425 430 Leu Glu Ser Val Val Ser Arg Thr Ser Pro His Gly Ile Leu Asp Ile 435 440 445 Leu Val His Glu Arg Ser Leu Asn Arg Gly Val Pro Pro Ser Tyr Pro 450 455 460 Pro Pro Glu Gln Phe Phe Asn Pro Ser Thr Tyr Asn Phe Asp Ala Thr 465 470 475 480 Cys Leu Leu Asn Ser Gly Val Val His Leu Thr Cys Thr Gly Phe Gln 485 490 495 Lys Glu Thr Leu Thr Phe Leu Lys Lys Ser Gly Pro Ser Leu Asn Glu 500 505 510 Leu Ile Pro Asp Gly Tyr Asn Arg Cys Leu Val Ala Gly Leu Leu Ser 515 520 525 Pro Arg Phe Val Asp Val Gln Pro Ser Ser Leu Ser Gln Glu Glu Gln 530 535 540 Leu Glu Ala Ile Leu Ser Ala Ala Ile Gln Thr Ser Ser Leu Gly Leu 545 550 555 560 Leu Thr Gly Tyr Ile Arg Arg Trp Ile Thr Glu Glu Gln Pro Asn Ser 565 570 575 Ala Thr Asn Leu Arg Phe Val Leu Glu Trp Thr Trp Asn Lys Val Val 580 585 590 Leu Thr Lys Glu Glu Phe Asp Arg Leu Cys Val Pro Leu Phe Asp Gly 595 600 605 Ser Cys His Phe Met Asp Pro Gln Thr Ile Gln Ser Ile Gln Gln Cys 610 615 620 Tyr Leu Leu Leu Ser Asn Leu Asn Ile Val Leu Ser Cys Phe Ala Ser 625 630 635 640 Glu Ala Arg Glu Ile Ala Glu Arg Gly Leu Ile Asp Leu Ser Asn Lys 645 650 655 Phe Val Val Ser His Leu Ile Cys Gln Tyr Ala Gln Val Val Leu Trp 660 665 670 Phe Ser His Ser Gly Leu Leu Pro Glu Gly Ile Asp Asp Ser Val Gln 675 680 685 Leu Ser Arg Leu Cys Tyr Asn Tyr Pro Val Ile Gln Asn Tyr Tyr Thr 690 695 700 Ser Arg Arg Gln Lys Phe Glu Arg Leu Ser Arg Gly Lys Trp Asn Pro 705 710 715 720 Asp Cys Leu Met Ile Asp Gly Leu Val Ser Gln Leu Gly Glu Arg Ile 725 730 735 Glu Lys Leu Trp Lys Arg Asp Glu Gly Gly Thr Gly Lys Tyr Pro Pro 740 745 750 Ala Ser Leu His Ala Val Leu Asp Met Tyr Leu Leu Asp Gly Val Thr 755 760 765 Glu Ala Ala Lys His Ser Ile Thr Ile Tyr Leu Leu Leu Asp Ile Met 770 775 780 Tyr Ser Phe Pro Asn Lys Thr Asp Thr Pro Ile Glu Ser Phe Pro Thr 785 790 795 800 Val Phe Ala Ile Ser Trp Gly Gln Val Lys Leu Ile Gln Gly Phe Trp 805 810 815 Leu Ile Asp His Asn Asp Tyr Glu Ser Gly L eu Asp Leu Leu Phe His 820 825 830 Pro Ala Thr Ala Lys Pro Leu Ser Trp Gln His Ser Lys Ile Ile Gln 835 840 845 Ala Phe Met Ser Gln Gly Glu His Arg Gln Ala Leu Arg Tyr Ile Gln 850 855 860 Thr Met Lys Pro Thr Val Ser Ser Gly Asn Asp Val Ile Leu His Leu 865 870 875 880 Thr Val Leu Leu Phe Asn Arg Cys Met Val Glu Ala Trp Asn Phe Leu 885 890 895 Arg Gln His Cys Asn Arg Leu Asn Ile Glu Glu Leu Leu Lys His Met 900 905 910 Tyr Glu Val Cys Gln Glu Met Gly Leu Met Glu Asp Leu Leu Lys Leu 915 920 925 Pro Phe Thr Asp Thr Glu Gln Glu Cys Leu Val Lys Phe Leu Gln Ser 930 935 940 Ser Ala Ser Val Gln Asn His Glu Phe Leu Leu Val His His Leu Gln 945 950 955 960 Arg Ala Asn Tyr Val Pro Ala Leu Lys Leu Asn Gln Thr Leu Lys Ile 965 970 975 Asn Val Met Asn Asp Arg Asp Pro Arg Leu Arg Glu Arg Ser Leu Ala 980 985 990 Arg Asn Ser Ile Leu Asp Gln Tyr Gly Lys Ile Leu Pro Arg Val His 995 1000 1005 Arg Lys Leu Ala Ile Glu Arg Ala Lys Pro Tyr His Leu Ser Thr Ser 1010 1015 1020 Ser Val Phe Arg Leu Val Ser Arg Pro Lys Pro Leu Ser Ala Val Pro 1025 1030 1035 1040 Lys Gln Val Val Thr Gly Thr Val Leu Thr Arg Ser Val Phe Ile Asn 1045 1050 1055 Asn Val Leu Ser Lys Ile Gly Glu Val Trp Ala Ser Lys Glu Pro Ile 1060 1065 1070 Asn Ser Thr Thr Pro Phe Asn Ser Ser Lys Ile Glu Glu Pro Ser Pro 1075 1080 1085 Ile Val Tyr Ser Leu Pro Ala Pro Glu Leu Pro Glu Ala Phe Phe Gly 1090 1095 1100 Thr Pro Ile Ser Lys Ala Ser Gln Lys Ile Ser Arg Leu Leu Asp Leu 1105 1110 1115 1120 Val Val Gln Pro Val Pro Arg Pro Ser Gln Cys Ser Glu Phe Ile Gln 1125 1130 1135 Gln Ser Ser Met Lys Ser Pro Leu Tyr Leu Val Ser Arg Ser Leu Pro 1140 1145 1150 Ser Ser Ser Gln Leu Lys Gly Ser Pro Gln Ala Ile Ser Arg Ala Ser 1155 1160 1165 Glu Leu His Leu Leu Glu Thr Pro Leu Val Val Lys Lys Ala Lys Ser 1170 1175 1180 Leu Ala Met Ser Val Thr Thr Ser Gly Phe Ser Glu Phe Thr Pro Gln 1185 1190 1195 1200 Ser Ile Leu Arg Ser Thr Pro Arg Ser Thr Pro Leu Ala Ser Pro Ser 1205 1210 1215 Pro Ser Pro Gly Arg Ser Pro Gln Arg Leu Lys Glu Thr Arg Ile Ser 1220 1225 1230 Phe Val Glu Glu Asp Val His Pro Lys Trp Ile Pro Gly Ala Ala Asp 1235 1240 1245 Asp Ser Lys Leu Glu Val Phe Thr Thr Pro Lys Lys Cys Ala Val Pro 1250 1255 1260 Val Glu Thr Glu Trp Pro Lys Ser Lys Asp Arg Thr Thr Ser Phe Phe 1265 1270 1275 1280 Leu Asn Ser Pro Glu Lys Glu His Gln Glu Met Asp Glu Gly Ser Gln 1285 1290 1295 Ser Leu Glu Lys Leu Asp Val Ser Lys Gly Asn Ser Ser Val Ser Ile 1300 1305 1310 Thr Ser Asp Glu Thr Thr Leu Glu Tyr Gln Asp Ala Pro Ser Pro Glu 1315 1320 1325 Asp Leu Glu Glu Thr Val Phe Thr Ala Ser Lys Pro Lys Ser Ser Ser 1330 1335 1340 Thr Ala Leu Thr Thr Asn Val Thr Glu Gln Thr Glu Lys Asp Gly Asp 1345 1350 1355 1360 Lys Asp Val Phe Ala Ser Glu Val Thr Pro Ser Asp Leu Gln Lys Gln 1365 1370 1375 Met Gly Asn Leu Glu Asp Ala Glu Thr Lys Asp Leu Leu Val Ala Ala 1380 1385 1390 Glu Ala Phe Ser Glu Leu Asn His Leu Ser Pro Val Gln Gly Thr Glu 1395 1400 1405 Ala Ser Leu Cys Ala Pro Ser Val Tyr Glu Gly Lys Ile Phe Thr Gln 1410 1415 1420 Lys Ser Lys Val Pro Val Leu Asp Glu Gly Leu Thr Ser Val Glu Thr 1425 1430 1435 1440 Tyr Thr Pro Ala Ile Arg Ala Asn Asp Asn Lys Ser Met Ala Asp Val 1445 1450 1455 Leu Gly Asp Gly Gly Asn Ser Ser Leu Thr Ile Ser Glu Gly Pro Ile 1460 1465 1470 Val Ser Glu Arg Arg Leu Asn Gln Glu Val Ala Leu Asn Leu Lys Glu 1475 1480 1485 Asp His Glu Val Glu Val Gly Val Leu Lys Glu Ser Val Asp Leu Pro 1490 1495 1500 Glu Glu Lys Leu Pro Ile Ser Asp Ser Pro Pro Asp Thr Gln Glu Ile 1505 1510 1515 1520 His Val Ile Glu Gln Glu Lys Leu Glu Ala Gln Asp Ser Gly Glu Glu 1525 1530 1535 Ala Arg Asn Leu Ser Phe Asn Glu Leu Tyr Pro Ser Gly Thr Leu Lys 1540 1545 1550 Leu Gln Tyr Asn Phe Asp Thr Ile Asp Gln Gln Phe Cys Asp Leu Ala 1555 1560 1565 Asp Asn Lys Asp Thr Ala Glu Cys Asp Ile Ala Glu Val Asp Gly Glu 1570 1575 1580 Leu Phe Val Ala Gln Ser Asn Phe Thr Leu Ile Leu Glu Gly Glu Glu 1585 1590 1595 1600 Gly Glu Val Glu Pro Gly Asp Phe Ala Ser Ser Asp Val Leu Pro Lys 1605 1610 1615 Ala Ala Asn Thr Ala Thr Glu Glu Lys Leu Val Cys Ser Gly Glu Glu Asn 1620 1625 1630 Asp Asn His Gly Gln Ile Ala Asn Leu Pro Ser Ala Val Thr Ser Asp 1635 1640 1645 Gln Lys Ser Gln Lys Val Asp Thr Leu Pro Tyr Val Pro Glu Pro Ile 1650 1655 1660 Lys Val Ala Ile Ala Glu Asn Leu Leu Asp Val Ile Lys Asp Thr Arg 1665 1670 1675 1680 Ser Lys Glu Ile Thr Ser Asp Thr Met Glu Gln Ser Ile His Glu Thr 1685 1690 1695 Ile Pro Leu Val Ser Gln Asn Ile Met Cys Pro Thr Lys Leu Val Lys 1700 1705 1710 Ser Ala Phe Lys Thr Ala Gln Glu Thr Ser Thr Met Thr Met Asn Val 1715 1720 1725 Ser Gln Val Asp Asp Val Val Ser Ser Lys Thr Arg Thr Arg Gly Gln 1730 1735 1740 Arg Ile Gln Asn Val Asn Val Lys Ser Ala Gln Gln Glu Ala Ser Ala 1745 1750 1755 1760 Asp Val Ala Thr Pro Lys Met Pro Gly Gln Ser Val Arg Lys Lys Thr 1765 1770 1775 Arg Lys Ala Lys Glu Ile Ser Glu Ala Ser Glu Asn Ile Tyr Ser Asp 1780 1785 1790 Val Arg Gly Leu Phe Gln Asn Gln Gln Ile Pro Gln Asn Ser Val Thr 1795 1800 1805 Pro Arg Arg Gly Arg Arg Lys Lys Glu Val Asn Gln Asp Ile L eu Glu 1810 1815 1820 Asn Thr Ser Ser Val Glu Gln Glu Leu Gln Ile Thr Thr Gly Arg Glu 1825 1830 1835 1840 Ser Lys Arg Leu Lys Ser Ser Gln Leu Leu Glu Pro Ala Val Glu Glu 1845 1850 1855 Thr Thr Lys Lys Glu Val Lys Val Ser Ser Val Thr Lys Arg Thr Pro 1860 1865 1870 Arg Arg Ile Lys Arg Ser Val Glu Asn Gln Glu Ser Val Glu Ile Ile 1875 1880 1885 Asn Asp Leu Lys Val Ser Thr Val Thr Ser Pro Ser Arg Met Ile Arg 1890 1895 1900 Lys Leu Arg Ser Thr Asn Leu Asp Ala Ser Glu Asn Thr Gly Asn Lys 1905 1910 1915 1920 Gln Asp Asp Lys Ser Ser Asp Lys Gln Leu Arg Ile Lys His Val Arg 1925 1930 1935 Arg Val Arg Gly Arg Glu Val Ser Pro Ser Asp Val Arg Glu Asp Ser 1940 1945 1950 Asn Leu Glu Ser Ser Gln Leu Thr Val Gln Ala Glu Phe Asp Met Ser 1955 1960 1965 Ala Ile Pro Arg Lys Arg Gly Arg Pro Arg Lys Ile Asn Pro Ser Glu 1970 1975 1980 Asp Val Gly Ser Lys Ala Val Lys Glu Glu Arg Ser Pro Lys Lys Lys 1985 1990 1995 2000 Glu Ala Pro Ser Ile Arg Arg Arg Ser Thr Arg Asn Thr Pro Ala Lys 2005 2010 2015 Ser Glu Asn Val Asp Val Gly Lys Pro Ala Leu Gly Lys Ser Ile Leu 2020 2025 2030 Val Pro Asn Glu Glu Leu Ser Met Val Met Ser Ser Lys Lys Lys Leu 2035 2040 2045 Thr Lys Lys Thr Glu Ser Gln Ser Gln Lys Arg Ser Leu His Ser Val 2050 2055 2060 Ser Glu Glu Arg Thr Asp Glu Met Thr His Lys Glu Thr Asn Glu Gln 2065 2070 2075 2080 Glu Glu Glu Arg Leu Leu Ala Thr Ala Ser Phe Thr Lys Ser Ser Arg Ser 2085 2090 2095 Ser Arg Thr Arg Ser Ser Lys Ala Ile Leu Leu Pro Asp Leu Ser Glu 2100 2105 2110 Pro Asn Asn Glu Pro Leu Phe Ser Pro Ala Ser Glu Val Pro Arg Lys 2115 2120 2125 Ala Lys Ala Lys Lys Ile Glu Val Pro Ala Gln Leu Lys Glu Leu Val 2130 2135 2140 Ser Asp Leu Ser Ser Gln Phe Val Ile Ser Pro Pro Ala Leu Arg Ser 2145 2150 2155 2160 Arg Gln Lys Asn Thr Ser Asn Lys Asn Lys Leu Glu Asp Glu Leu Lys 2165 2170 2175 Asp Asp Ala Gln Ser Val Glu Thr Leu Gly Lys Pro Lys Ala Lys Arg 2180 2185 2190 Ile Arg Thr Ser Lys Thr Lys Gln Ala Ser Lys Asn Thr Glu Lys Glu 2195 2200 2205 Ser Ala Trp Ser Leu Pro Pro Ile Glu Ile Arg Leu Ile Ser Pro Leu 2210 2215 2220 Ala Ser Pro Ala Asp Gly Val Lys Ser Lys Pro Arg Lys Thr Thr Glu 2225 2230 2235 2240 Val Thr Gly Thr Gly Leu Gly Arg Asn Arg Lys Lys Leu Ser Ser Tyr 2245 2250 2255 Pro Lys Gln Ile Leu Arg Arg Lys Met Leu 2260 2265 <210> 15 <211> 7215 <212> DNA <213> Homo sapiens <220> <221> CDS <222> (98) .. (6922) <400> 15 tgcggctcga gggggccagc gctgacggtg gcgggtacgg caggctcgcg ggcgccgggc 60 ttcgttacat aatctcggac cggaggagcg gtggcac atg gc g cg gaa cgg cgc gat gag gag gag gag gag gag gag gag gag gag gag gag gt g Met Arg Asp Leu Arg Ala Gln Val Thr Ser Gly Leu Leu 10 15 20 cca ttt cca gaa gtg act ctt caa gcc ctt gga gaa gac gaa ata aca 211 Pro Phe Pro Glu Val Thr Leu Gln Ala Leu Gly Glu Asp Glu Ile Thr 25 30 35 tta gaa tct gtg ctt cgt gga aag ttt gct gcg ggg aaa aat gga ctt 259 Leu Glu Ser Val Leu Arg Gly Lys Phe Ala Ala Gly Lys Asn Gly Leu 40 45 50 gct tgc ttg gct tgt ggt cca caa ctt gag gta aac tct ata aca 307 Ala Cys Leu Ala Cys Gly Pro Gln Leu Glu Val Val Asn Ser Ile Thr 55 60 65 70 gga gag cga ttg tct gct tac aga ttc agt gga gtc aat gaa cag cct 355 Gly Glu Arg Leu Ser Ala Tyr Arg Phe Ser Gly Val Asn Glu Gln Pro 75 80 85 cct gta gtt tta gct gtg aaa gaa ttc tct tgg cag aag aga act gga 403 Pro Val Val Leu Ala Val Lys Glu Phe Ser T rp Gln Lys Arg Thr Gly 90 95 100 tta tta ata gga ttg gaa gaa aca gaa ggg agt gtt ctc tgt ctt tat 451 Leu Leu Ile Gly Leu Glu Glu Thr Glu Gly Ser Val Leu Cys Leu Tyr 105 110 115 gac ctt gga ata tca aaa gta gtt aaa gca gtt gtt ctt cct gga agg 499 Asp Leu Gly Ile Ser Lys Val Val Lys Ala Val Val Leu Pro Gly Arg 120 125 130 gta aca gct att gaa cct ata att aat cat gga gga gcc agt gca agc 547 Val Thr Ala Ile Glu Pro Ile Ile Asn His Gly Gly Ala Ser Ala Ser 135 140 145 150 act cag cat tta cat cca agt ctg cga tgg ctt ttt gga gtg gca gct 595 Thr Gln His Leu His Pro Ser Leu Arg Trp Leu Phe Gly Val Ala Ala 155 160 165 gtg gtc act gat gtt gga cag atc ctt ctt att gac cta tgt ttg gat 643 Val Val Thr Asp Val Gly Gln Ile Leu Leu Ile Asp Leu Cys Leu Asp 170 175 180 gac ttg tca tgc aat caa aat gaa gtt ga gca tca gat ctt gaa gtt 691 Asp Leu Ser Cys Asn Gln Asn Glu Val Glu Ala Ser Asp Leu Glu Val 185 190 195 cta act ggt atc cca gct gaa gta cca cac att aga gaa agt gtg atg 739 Leu Thr Gly Ile Pro Ala Glu Val Pro His Ile Arg Glu Ser Val Met 200 205 210 aga gaa ggg cgc cat ctg tgt ttc cag tta gta agt cca aca gga aca 787 Arg Glu Gly Arg His Leu Cys Phe Gln Leu Val Ser Pro Thr Gly Thr 215 220 225 230 gcc gtt tca act ctt agt tac ata agc agg aca aat cag ctt gct gca 835 Ala Val Ser Thr Leu Ser Tyr Ile Ser Arg Thr Asn Gln Leu Ala Ala 235 240 245 ggt ttt tct gat ggc tat cta gca ctt tgg aac atg aaa agc atg aaa 883 Gly Phe Ser Asp Gly Tyr Leu Ala Leu Trp Asn Met Lys Ser Met Lys 250 255 260 aga gaa tat tac ata caa ttg gaa agt gga caa gtt cct gta tat gct 931 Arg Glu Tyr Tyr Ile Gln Leu Glu Ser Gly Gln Val Pro Val Tyr Ala 265 270 275 gtc act ttt caa gaa cct gag aat gat cgt cgg aat tgc tgc tac ttg 979 Val Thr Phe Gln Glu Pro Glu Asn Asp Arg Arg Asn Cys Cys Tyr Leu 280 285 290 290 tgg gct gtt cag tca caa gat agt gaa ggg gat gtt ttg agt ttg 1027 Trp Ala Val Gln Ser Thr Gln Asp Ser Glu Gly Asp Val Leu Ser Leu 295 300 305 310 cat ctg ctg cag ctg gcc ttt ggt aat aga aag tgt ttg gca tca gga 1075 His Leu Leu Gln Leu Ala Phe Gly Asn Arg Lys Cys Leu Ala Ser Gly 315 320 325 caa atc tta tat gag ggg tta gaa tac tgt gaa gaa aga tac acc ctg 1123 Gln Ile Leu Tyr Glu Gly Leu Glu Tyr Cys Glu Glu Arg Thr Leu 330 335 340 gac ctg aca ggt ggc atg ttc cct ttg agg gga cag acg agt aat acc 1171 Asp Leu Thr Gly Gly Met Phe Pro Leu Arg Gly Gln Thr Ser Asn Thr 345 350 355 aaa ttg ttg gga tgc cag agt atgag aaa ttt cga tct cat ggt gac 1219 Lys Leu Leu Gly Cys Gln Ser Ile Glu Lys Phe Arg Ser His Gly Asp 360 365 370 agg gag gaa ggc gtg aat gaa gct cta tcg cct gac act agt gtt tca 1267 Arg Glu Glu Gly Val Asn Glu Ala Leu Ser Pro Asp Thr Ser Val Ser 375 380 385 390 gtc ttt acc tgg cag gtg aat ata tat gga cag gga aag cct tct gta 1315 Val Phe Thr Trp Grp Val Asn Ile Tyr Gly Gln Gly Lys Pro Ser Val 395 400 405 tat ttg ggg ctt ttt gat ata aat cgt tgg tat cat gca caa atg cca 1363 Tyr Leu Gly Leu Phe Asp Ile Asn Arg Trp Tyr His Ala Gln Met Pro 410 415 420 gat tcg tta agg tca gga gaa tat cta cat aa t tgc tct tat ttt gca 1411 Asp Ser Leu Arg Ser Gly Glu Tyr Leu His Asn Cys Ser Tyr Phe Ala 425 430 435 ctg tgg tca ttg gag tct gtt gta agt agg act tct cca cat ggc atc 1459 Leu Trp Ser Leu Glu Ser Val Val Ser Arg Thr Ser Pro His Gly Ile 440 445 450 ttg gat ata tta gta cat gag aga agt tta aat aga gga gtc cct cct 1507 Leu Asp Ile Leu Val His Glu Arg Ser Leu Asn Arg Gly Val Pro Pro 455 460 465 465 470 tca tat cca cct ccc gag cag ttt ttt aat cca agc act tat aat ttt 1555 Ser Tyr Pro Pro Glu Gln Phe Phe Asn Pro Ser Thr Tyr Asn Phe 475 480 485 gat gcc act tgt ttg tta aac tcg gga gtt gtt cat tta act tgt act 1603 Asp Ala Thr Cys Leu Leu Asn Ser Gly Val Val His Leu Thr Cys Thr 490 495 500 ggc ttt cag aag gag act ttg act ttt tta aag aaa tca ggt cca tca 1651 Gly Phe Gln Lys Glu Thr Leu Thr Phe Leu Lys Lys Ser Gly Pro Ser 505 510 515 ctc at gaa ctc att cct gat ggt tat aat cga tgt ctt gta gct ggc 1699 Leu Asn Glu Leu Ile Pro Asp Gly Tyr Asn Arg Cys Leu Val Ala Gly 520 525 530 530 ctt ctt tcc cca aga ttt gtt gat gtt cag cct tcc agt tta agc caa 1747 Leu Leu Ser Pro Arg Phe Val Asp Val Gln Pro Ser Ser Leu Ser Gln 535 540 545 550 gaa gaa cag tta gaa gct ata ttg tca gca gca att cag act agt tcc 1795 Glu Glu Gln Leu Glu Ala Ile Leu Ser Ala Ala Ile Gln Thr Ser Ser 555 560 565 ctg gga ctt ttg act ggt tat atc cga aga tgg ata aca gaa gaa caa 1843 Leu Gly Leu Leu Thr Gly Tyr Ile Arg Arg Trp Ile Thr Glu Gln 570 575 580 cca aat tct gcc act aat ttg cgc ttt gtt ctt gaa tgg acg tgg aat 1891 Pro Asn Ser Ala Thr Asn Leu Arg Phe Val Leu Glu Trp Thr Trp Asn 585 590 595 aaa gtg gtt ctc aca tt gagga gac aga cta tgt gtg cca tta 1939 Lys Val Val Leu Thr Lys Glu Glu Phe Asp Arg Leu Cys Val Pro Leu 600 605 610 ttt gat ggt tcg tgt cat ttc atg gat cca caa act ata cag tct atc 1987 Phe Asp Gly Ser Cys His Phe Met Asp Pro Gln Thr Ile Gln Ser Ile 615 620 625 630 cag caa tgc tat ttg ctt ctt agc aat ctt aat ata gtc ttg agc tgt 2035 Gln Gln Gln Cys Tyr Leu Leu Leu Ser Asn Leu Asn Ile Val Leu Ser Cy s 635 640 645 ttt gca tca gaa gcc cga gag atc gct gag aga gga ctg ata gac tta 2083 Phe Ala Ser Glu Ala Arg Glu Ile Ala Glu Arg Gly Leu Ile Asp Leu 650 655 660 agc aat aag ttt gtg gtt tcc tgt cag tat gca caa gtg 2131 Ser Asn Lys Phe Val Val Ser His Leu Ile Cys Gln Tyr Ala Gln Val 665 670 675 gtt ctt tgg ttc tct cat tct ggg ctt tta cca gaa ggc ata gat gat 2179 Val Leu Trp Phe Ser His Ser Gly Leu Leu Pro Glu Gly Ile Asp Asp 680 685 690 tct gtg cag ttg tca agg tta tgc tac aac tac cct gta att cag aac 2227 Ser Val Gln Leu Ser Arg Leu Cys Tyr Asn Tyr Pro Val Ile Gln Asn 695 700 705 710 tac tac acc agt cgt cga cag aag ttt gag cgt tta tca aga ggg aag 2275 Tyr Tyr Thr Ser Arg Arg Gln Lys Phe Glu Arg Leu Ser Arg Gly Lys 715 720 725 tgg aat ccc gat tgc ttg atg att gat gga ctg gtt tct cag gga 2323 Trp Asn Pro Asp Cys Leu Met Ile Asp Gly Leu Val Ser Gln Leu Gly 730 735 740 gag cga att gag aag ttg tgg aaa cga gat gaa gga ggc aca gga aaa 2371 Glu Arg Ile Glu Lys Leu Trp Lys Arg Asp Glu Gly Gly Thr Gly Lys 745 750 755 tat cct cct gct agt ctg cat gca gta ctt gat atg tac cta tta gac 2419 Tyr Pro Pro Ala Ser Leu His Ala Val Leu Asp Met Tyr Leu Leu Asp 760 765 770 ggc gtt act gaa gca gcc aaa cac tct att acc att tat ttg cta ctt 2467 Gly Val Thr Glu Ala Ala Lys His Ser Ile Thr Ile Tyr Leu Leu Leu 775 780 785 790 gat att atg tat tcc ttt ccc aac aaa aca gac act ccc att gaa tct 2515 Asp Ile Met Tyr Ser Phe Pro Asn Lys Thr Asp Thr Pro Ile Glu Ser 795 800 805 ttc cca act gta ttt gcc att tct tgg ggc caa gtt aaa ctt att cag 2563 Phe Pro Thr Val Phe Ala Ile Ser Trp Gly Gln Val Lys Leu Ile Gln 810 815 820 ggg ttt tgg ttg ata gat cat aat gac tat gag agt ggt ttg gat ctt 2611 Gly Phe Trp Leu Ile Asp His Asn Asp Tyr Glu Ser Gly Leu Asp Leu 825 830 835 ttg ttt cat cca gct act gca aaac ttg tca tgg caa cat tca aag 2659 Leu Phe His Pro Ala Thr Ala Lys Pro Leu Ser Trp Gln His Ser Lys 840 845 850 att att cag gca ttc atg agt cag ggc gag cac aga caa gcc ctc aga 2707 Ile Ile Gln Ala Phe Met Ser Gln Gly Glu His Arg Gln Ala Leu Arg 855 860 865 870 tat att cag aca atg aag cca aca gtg tcc agt ggt aac gat gtt atc 2755 Tyr Ile Gln Thr Met Lys Pro Thr Val Ser Ser Gly Asn Asp Val Ile 875 880 885 ctt cac ctc act gtt ttg ctt ttt aat agg tgt atg gtt gaa gcc tgg 2803 Leu His Leu Thr Val Leu Leu Phe Asn Arg Cys Met Val Glu Ala Trp 890 895 900 aat ttt ttg cgg caa cat tgc aat agg ttg ata gag gag tta ctg 2851 Asn Phe Leu Arg Gln His Cys Asn Arg Leu Asn Ile Glu Glu Leu Leu 905 910 915 aag cac atg tat gaa gtc tgt cag gaa atg ggc ttg atg gaa gat tta 2899 Lys His Met Tyr Gyr Glu Met Gly Leu Met Glu Asp Leu 920 925 930 ctg aag tta cca ttt aca gac act gag cag gaa tgt tta gtg aaa ttt 2947 Leu Lys Leu Pro Phe Thr Asp Thr Glu Gln Glu Cys Leu Val Lys Phe 935 940 940 950 g tcc agt gcc agc gtt cag aat cat gaa ttc ctt tta gtg cac 2995 Leu Gln Ser Ser Ala Ser Val Gln Asn His Glu Phe Leu Leu Val His 955 960 965 cat ttg cag cgt gcc aat tat gtg cct gcc ttg aag ct g aac caa act 3043 His Leu Gln Arg Ala Asn Tyr Val Pro Ala Leu Lys Leu Asn Gln Thr 970 975 980 ctg aag att aat gtt atg aat gat cgt gat cct cgt ttg cgg gag aga 3091 Leu Lys Ile Asn Val Met Asn Asp Arg Asp Pro Arg Leu Arg Glu Arg 985 990 995 tca ctg gct cga aat tct ata tta gac cag tat gga aaa atc ctt cct 3139 Ser Leu Ala Arg Asn Ser Ile Leu Asp Gln Tyr Gly Lys Ile Leu Pro 1000 1005 1010 aga gtc cat cga aaa tta gcc att gaa cga gct aag cct tat cat ctg 3187 Arg Val His Arg Lys Leu Ala Ile Glu Arg Ala Lys Pro Tyr His Leu 1015 1020 1025 1030 tca aca tca tca gtt ttt cga tta gtt tct aga ccc aaa cca tta tca 3235 Ser Thr Ser Ser Val Phe Arg Leu Val Ser Arg Pro Lys Pro Leu Ser 1035 1040 1045 gca gtt cca aag caa gtt gta aca gga act gtg ttg aca aga tct gtt 3283 Ala Val Pro Lys Gln Val Val Thr Gly Thr Val Leu Thr Arg Ser Val 1050 1055 1060 ttc atc aac aat gtg tta tct aaa att gga gaa gtt tgg gca agc aaa 3331 Phe Ile Asn Asn Val Leu Ser Lys Ile Gly Glu Val Trp Ala Ser Lys 1065 1070 1075 gaa cct ata aat agc acc aca cct ttc aat agt tct aaa ata gaa gaa 3379 Glu Pro Ile Asn Ser Thr Thr Pro Phe Asn Ser Ser Lys Ile Glu Glu 1080 1085 1090 cca tct cct ata gtg tat tcg ctc cca gct cca gag ctg cct gag 3427 Pro Ser Pro Ile Val Tyr Ser Leu Pro Ala Pro Glu Leu Pro Glu Ala 1095 1100 1105 1110 ttt ttt gga aca cca att tca aaa gca tca caa aaa att tct aga ctg 3475 Phe Phe Gly Thr Pro Ile Ser Lys Ala Ser Gln Lys Ile Ser Arg Leu 1115 1120 1125 cta gat ttg gtt gtt cag cct gtc ccc cgg cct tct cag tgt tcg gag 3523 Leu Asp Leu Val Val Gln Pro Val Pro Arg Pro Ser Gln Cys Ser Glu 1130 1135 1140 ttt att cag caa agc tcc atg aaa tct cct ttg tac cta gta tcc cgt 3571 Phe Ile Gln Gln Ser Ser Met Lys Ser Pro Leu Tyr Leu Val Ser Arg 1145 1150 1155 tca ctg ccc tca agt tcg caa tta aaa gga tcg cct cag gcc atc tcc 3619 Ser Leu Pro Ser Ser Ser Gln Leu Lys Gly Ser Pro Gln Ala Ile Ser 1160 1165 1170 agg gct tca gaa tta cat ttg ctt gaa act cct ctt gta gtt aag aaa 3667 Arg Ala Ser Glu Leu His Leu Leu Glu Thr Pro Leu Val Val Lys Lys 1175 1180 1185 1190 gct aaa agt ttg gcc atg tca gtt act act tct gga ttt tct gag ttc 3715 Ala Lys Ser Leu Ala Met Ser Val Thr Thr Ser Gly Phe Ser Glu Phe 1195 1200 1205 act cct cag tcc atc ctg agg tct act cct cga tca aca cct tta gca 3763 Thr Pro Gln Ser Ile Leu Arg Ser Thr Pro Arg Ser Thr Pro Leu Ala 1210 1215 1220 tct ccc tct cca tca cct gga agg tct cct caa cga ctt aaa gaa act 3811 Ser Pro Ser Pro Ser Pro Gly Arg Ser Pro Gln Arg Leu Lys Glu Thr 1225 1230 1235 aga att tca ttt gtg gaa gaa gat gtc cac cca aaa tgg att cct ggg 3859 Arg Ile Ser Phe Val Glu Glu Asp Val His Pro Lys Trp Ile Pro Gly 1240 1245 1250 gct gca gat gat agc aaa tta gaa gta ttt act aca cct aaa aaa tgt 3907 Ala Ala Asp Asp Ser Lys Leu Glu Val Phe Thr Thr Pro Lys Lys Cys 1255 1260 1265 1270 gca gtt cca gtg gaa act gga tgg aag agc aaa gat agg acc aca 3955 Ala Val Pro Val Glu Thr Glu Trp Pro Lys Ser Lys Asp Arg Thr Thr 1275 1280 1285 tct ttt ttc ctg aac agc cct gaa aag gag cat caa gaa atg gat gag 4003 Ser Phe Phe Leu Asn Ser Pro Glu Lys Glu His Gln Glu Met Asp Glu 1290 1295 1300 ggg tca caa agt tta gag aaa ctg gat gtg agc aaa gga aac agc agt 4051 Gly Ser Gln Ser Leu Glu Lys Leu Asp Val Ser Lys Gly Asn Ser Ser 1305 1310 1315 gtt tca atc aca tcc gat gag act acc tta gag tat cag gat gca ccg 4099 Val Ser Ile Thr Ser Asp Glu Thr Thr Leu Glu Tyr Gln Asp Ala Pro 1320 1325 1330 tca ccg gaa gac ctt gaa gag act gtt ttc acg gcc tct aag ccc aaa 4147 Ser Pro Glu Asp Leu Glu Glu Thr Val Phe Thr Ala Ser Lys Pro Lys 1335 1340 1345 1350 agc tct tcc act gca cta act act aat gta act gaa caa act gaa aag 4195 Ser Ser Ser Thr Ala Leu Thr Thr Asn Val Thr Glu Gln Thr Glu Lys 1355 1360 1365 gat gga gat aaa gat gta ttt gca tca gaa gta act cct tca gac cta 4243 Asp Gly Asp Lys Asp Val Phe Ala Ser Glu Val Thr Pro Ser Asp Leu 1370 1375 1380 cag aaa caa atg ggc aat tta gaa gat gca gaa aca aag gat ctc tta 4291 Gln Lys Gln Met Gly Asn Leu Glu Asp Ala Glu Thr Lys Asp Leu Leu 1385 1390 1395 gtt gca gca gag gca ttt tca gaa ttg aat cac tta agc ccg gtt caa 4339 Val Ala Ala Glu Ala Phe Ser Glu Leu Asn His Leu Ser Pro Val Gln 1400 1405 1410 gga act gaa gct tct ctt tgt gca cca tca gtc tat gaa ggg aaa 4387 Gly Thr Glu Ala Ser Leu Cys Ala Pro Ser Val Tyr Glu Gly Lys Ile 1415 1420 1425 1430 ttc acc cag aag tcc aag gta cca gtg ttg gac gaa gga tta aca tct 4435 Phe Thr Gln Lys Ser Lys Val Pro Val Leu Asp Glu Gly Leu Thr Ser 1435 1440 1445 gtt gaa acc tac acc cct gca att aga gca aat gac aat aaa tct atg 4483 Val Glu Thr Tyr Thr Pro Ala Ile Arg Ala Asn Asp Asn Lys Ser Met 1450 1455 1460 gct gat gtc ctt ggt gat ggt gga aac tcc tcg ctc act atc tct gaa 4531 Ala Asp Val Leu Gly Asp Gly Gly Asn Ser Ser Leu Thr Ile Ser Glu 1465 1470 1475 ggt cct att gtc tct gag cgc agg ctt aac cag gaa gta gcg ctg aac 4579 Gly I Ser Glu Arg Arg Leu Asn Gln Glu Val Ala Leu Asn 1480 1485 1490 tta aaa gaa gat cat gaa gta gaa gtt ggt gta cta aaa gaa agt gtt 4627 Leu Lys Glu Asp His Glu Val Glu Val Gly Val Leu Lys Glu Ser Val 1495 1500 1505 1510 gac tta cca gaa gaa aag ctt cca att tct gac agc cct cct gat act 4675 Asp Leu Pro Glu Glu Glu Lys Leu Pro Ile Ser Asp Ser Pro Pro Asp Thr 1515 1520 1525 caa gaa att cat ggt att gaa caa gaa aag ctt gaa gct caa gat tca 4723 Gln Glu Ile His Val Ile Glu Gln Glu Lys Leu Glu Ala Gln Asp Ser 1530 1535 1540 gga gaa gag gct agg aat ctt tca ttt aat gag tta tat ccc tct g Glu Glu Ala Arg Asn Leu Ser Phe Asn Glu Leu Tyr Pro Ser Gly 1545 1550 1555 aca ctt aag ctt cag tac aat ttt gat act att gac caa cag ttt tgt 4819 Thr Leu Lys Leu Gln Tyr Asn Phe Asp Thr Ile Asp Gln Gln Phe Cys 1560 1565 1570 gac tta gct gat aac aaa gac act gct gaa tgt gac att gct gaa gta 4867 Asp Leu Ala Asp Asn Lys Asp Thr Ala Glu Cys Asp Ile Ala Glu Val 1575 1580 1585 1590 gat ggg gaa ctt ttt gc gca aac ttt acc ttg ata ttg gaa 4915 Asp Gly Glu Leu Phe Val Ala Gln Ser Asn Phe Thr Leu Ile Leu Glu 1595 1600 1605 ggt gaa gaa gga gaa gtt gag cca ggt gat ttt gca tca tct gat gtg 4963 Gly Glu Glu Gly Glu Val Glu Pro Gly Asp Phe Ala Ser Ser Asp Val 1610 1615 1620 tta cct aaa gca gct aac aca gca act gaa gaa aaa ctt gta tgc agt 5011 Leu Pro Lys Ala Ala Ala Asn Thr Ala Thr Glu Glu Lys Leu Val Cys Ser 1625 1630 1635 ggg gaa aat gat aat cat gga caa att gca aat ttg cca tct gcc gta 5059 Gly Glu Asn Asp Asn His Gly Gln Ile Ala Asn Leu Pro Ser Ala Val 1640 1645 1650 act agt gac caa aag tcc caa aaa gta gac act tta cca tat gtg cct 5107 Thr Ser Asp Gln Lys Ser Gln Lys Val Asp Thr Leu Pro Tyr Val Pro 1655 1660 1665 1670 gaa cct att aaa gta gca att gca gaa aat tta cta gat gta att aaa 5155 Glu Pro Ile Lys Val Ala Ile Ala Glu Asn Leu Leu Asp Val Ile Lys 1675 1680 1685 gac aca aga agt aaa gaa att act tca gat aca atg gaa cag tcc att 5203 Asp Thr Arg Ser Lys Glu Ile Thr Ser Asp Thr Met Glu Gln Ser Ile 1690 1695 1700 cat gaa aca ata cct tta gtg agc caa aac ata atg tgt ccc act aaa 5251 His Glu Thr Ile Pro Leu Val Ser Gln Asn Ile Met Cys Pro Thr Lys 1705 1710 1715 ttg gtc aaa tct gca ttt aag act gct cag gaa aca agc aca atg act 5299 Leu Val Lys Ser Ala Phe Lys Thr Ala Gln Glu Thr Ser Thr Met Thr 1720 1725 1730 atg aat gtc agc cag gtt gat gac gtg gtt tcc tcc aaa act cgt acg 5347 Met Asn Val Ser Gln Val Asp Asp Val Val Ser Ser Lys Thr Arg Thr 1735 1740 1745 1750 aga ggt caa cgt atc caa aac gtg aat gtc aaa tca gca caa cag gaa 5395 Arg Gly Gln Arg Ile Gln Asn Val Asn Val Lys Ser Ala Gln Gln Glu 1755 1760 1765 gca tca gca gat gtt gct act cct aag atg cca ggg cag tca gtc agg 5443 Ala Ser Ala Asp Val Ala Thr Pro Lys Met Pro Gly Gln Ser Val Arg 1770 1775 1780 aag aaa act agg aag gca aaa gaa att tct gaa gct tct gaa aac atc 5491 Lys Lys Thr Arg Lys Ala Lys Glu Ile Ser Glu Ala Ser Glu Asn Ile 1785 1790 1795 tat tct gat gtc aga gga cta ttt cag aac cag caa ata cct caa aat 5539 Tyr Serp Arg Gly Leu Phe Gln Asn Gln Gln Ile Pro Gln Asn 1800 1805 1810 tct gtt acg cct agg aga gga agg aga aag aaa gaa gtt aat cag gac 5587 Ser Val Thr Pro Arg Arg Gly Arg Arg Lys Lys Glu Val Asn Gln Asp 1815 1820 1825 1830 ata cta gaa aac acc agt tct gtg gaa caa gaa tta cag atc act aca 5635 Ile Leu Glu Asn Thr Ser Ser Val Glu Gln Glu Leu Gln Ile Thr Thr 1835 1840 1845 ggt agg gaa tca aaa aga tta aaa tca tct cag ctg ttg gaa cca gca 5683 Gly Arg Glu Ser Lys Arg Leu Lys Ser Ser Gln Leu Leu Glu Pro Ala 1850 1855 1860 gtt gaa gaa act act aaa aaa gaa gtt aag gtt tca tct gtt acaaaa 57 Glu Glu Thr Thr Lys Lys Glu Val Lys Val Ser Ser Val Thr Lys 1865 1870 1875 agg act cct aga aga att aaa aga tct gta gaa aat cag gaa agt gtt 5779 Arg Thr Pro Arg Arg Ile Lys Arg Ser Val Glu Asn Gln Glu Ser Val 1880 1885 1890 gaa att ata aat gat cta aaa gtt agt acg gta aca agt cct agc aga 5827 Glu Ile Ile Asn Asp Leu Lys Val Ser Thr Val Thr Ser Pro Ser Arg 1895 1900 1905 1910 atg atc aga aaa ttg aga agt act aat tta gat gct tct gaa aat aca 5875 Met Ile Arg Lys Leu Arg Ser Thr Asn Leu Asp Ala Ser Glu Asn Thr 1915 1920 1925 gga aat aag caa gat gat aaa tcc agt gac aag cag ctg cg t att aaa 5923 Gly Asn Lys Gln Asp Asp Lys Ser Ser Asp Lys Gln Leu Arg Ile Lys 1930 1935 1940 cat gtt aga agg gtc aga ggg aga gaa gtt agt cca tca gat gtg aga 5971 His Val Arg Arg Val Arg Gly Arg Glu Val Ser Pro Ser Asp Val Arg 1945 1950 1955 gaa gac tcc aac ctt gag tca tct cag ttg act gtt caa gca gaa ttt 6019 Glu Asp Ser Asn Leu Glu Ser Ser Gln Leu Thr Val Gln Ala Glu Phe 1960 1965 1970 gat atg tct gcc ata cct aga aaa cgt ggt aga cca aga aaa atc aat 6067 Asp Met Ser Ala Ile Pro Arg Lys Arg Gly Arg Pro Arg Lys Ile Asn 1975 1980 1985 1990 cca tct gaa gat gta gga tct aag gct gtt aag gaa gag aga agc ccc 6115 Ser Glu Asp Val Gly Ser Lys Ala Val Lys Glu Glu Glu Arg Ser Pro 1995 2000 2005 aag aag aaa gaa gct ccc agc att aga agg aga tct aca aga aat acc 6163 Lys Lys Lys Glu Ala Pro Ser Ile Arg Arg Arg Ser Thr Arg Asn Thr 2010 2015 2020 cca gct aaa agt gaa aat gtt gat gtt gga aaa cca gct tta gga aaa 6211 Pro Ala Lys Ser Glu Asn Val Asp Val Gly Lys Pro Ala Leu Gly Lys 2025 2030 2035 tcc at t tta gtg cca aac gag gaa ctt tcg atg gtg atg agc tct aag 6259 Ser Ile Leu Val Pro Asn Glu Glu Leu Ser Met Val Met Ser Ser Lys 2040 2045 2050 aaa aaa ctt aca aaa aag act gaa agt caa agc caa aaa cgt ttg 6307 Lys Lys Leu Thr Lys Lys Thr Glu Ser Gln Ser Gln Lys Arg Ser Leu 2055 2060 2065 2070 cac tca gta tca gaa gaa cgc aca gat gaa atg aca cat aaa gaa aca 6355 His Ser Val Ser Glu Glu Arg Thr Asp Glu Met Thr His Lys Glu Thr 2075 2080 2085 aat gag cag gaa gaa aga ttg ctc gcc aca gct tcc ttc act aaa tca 6403 Asn Glu Gln Glu Glu Arg Leu Leu Ala Thr Ala Ser Phe Thr Lys Ser 2090 2095 2100 tcc cgc agc agg agg cag cgg tct agc aag gcc atc ttg ttg ccg gac 6451 Ser Arg Ser Ser Arg Thr Arg Ser Ser Lys Ala Ile Leu Leu Pro Asp 2105 2110 2115 ctt tct gaa cca aac aat gag cct tta ttt tct cca gcg tca gaa gttlu Seru Pro Asn Asn Glu Pro Leu Phe Ser Pro Ala Ser Glu Val 2120 2125 2130 cca agg aaa gca aaa gct aaa aaa ata gag gtt cct gca cag ctg aaa 6547 Pro Arg Lys Ala Lys Ala Lys Lys Ile Gl u Val Pro Ala Gln Leu Lys 2135 2140 2145 2150 gaa tta gtt tcg gat tta tct tct cag ttt gtc atc tca cct cct gct 6595 Glu Leu Val Ser Asp Leu Ser Ser Gln Phe Val Ile Ser Pro Pro Ala 2155 2160 2165 tta agg agc aga caa aaa aac aca tcc aat aag aac aag ctt gaa gat 6643 Leu Arg Ser Arg Gln Lys Asn Thr Ser Asn Lys Asn Lys Leu Glu Asp 2170 2175 2180 gaa ctg aaa gat gat gca caa tca gta gaa act ctg gga aagca Glu Leu Lys Asp Asp Ala Gln Ser Val Glu Thr Leu Gly Lys Pro Lys 2185 2190 2195 gcg aaa cga atc agg acg tca aaa aca aaa caa gca agc aaa aac aca 6739 Ala Lys Arg Ile Arg Thr Ser Lys Thr Lys Gln Ala Ser Lys Asn Thr 2200 2205 2210 gaa aaa gaa agt gct tgg tca ctt cct ccc ata gaa att cgg ctg att 6787 Glu Lys Glu Ser Ala Trp Ser Leu Pro Pro Ile Glu Ile Arg Leu Ile 2215 2220 2225 2230 tcc ccc ttg gct agc cca gga gtc aag agc aaa cca aga aaa 6835 Ser Pro Leu Ala Ser Pro Ala Asp Gly Val Lys Ser Lys Pro Arg Lys 2235 2240 2245 act aca gaa gtg aca gga aca ggt ctt gga agg aac aga aag aaa ctg 6883 Thr Thr Glu Val Thr Gly Thr Gly Leu Gly Arg Asn Arg Lys Lys Leu 2250 2255 2260 tct tcc tat cca aag caa att tta cgc aga aaa atg ctg taatttcttg 6932 Ser Ser Tyr Pro Lys Gln Ile Leu Arg Leg Met 2265 2270 2275 ggaagatttt aatgtacacc tatttgtaaa gtcatcagaa tagtgtggat tattaaatat 6992 ctagtttgga agaaaataat ttatataaat tattgtaaat ttttatgtaa acagaaggtc 7052 ttcaataagt aaagtaactc catatggagt gattgtttca gtccaggcaa tttttctatt 7112 ttatattaag acttcataca tttatatatg taaatatggc ttattaatgg aatgttaaat 7172 aaaatgtata cttctcaaaa aaaaaaaaaa aaaaaaaaaa aaa 7215

[Brief description of the drawings]

FIG. 1 is a micrograph showing the localization of YS68 in cells. YS68 with a Flag epitope was expressed in COS7 cells, and the cells were stained with an anti-Flag antibody to examine the expression site (right). The same cells were treated with Hoechst to specifically stain the nuclei (left).

FIG. 2 is an electrophoretic photograph showing the expression distribution of YS68 in a tissue. Liver of mouse embryonic day 14 (E14), day 18 (E18),
RNA was prepared from each tissue of the spleen, thymus, or adult mouse, and Northern blot hybridization was performed. The lower panel shows 18S ribosomal R before blotting.
NA indicated as control.

Fig. 3 Expression of YS68 in the yolk sac at each stage of the fetus
4 is an electrophoretic photograph showing the result of analysis by RT-PCR.

FIG. 4 (A) is an electrophoretic photograph showing the results of RT-PCR analysis of YS68 expression in the AGM region at each stage of the fetus. (B) The AGM region of embryonic day 10.5 was transformed into Oncostatin M (OS
M) Culture in the presence or absence of RNA.
Was prepared. Then, the expression of YS68 was compared with the uncultured AGM region by RT-PCR.

[Fig. 5] Mouse embryo (11.5 to 16.5 days of embryo), postnatal
7 days, and extract RNA from adult liver, thymus, spleen,
4 is an electrophoresis photograph showing the result of comparing the expression level of YS68 by RT-PCR.

[Fig. 6] A section was prepared from a fetus on day 11.5 of fetal life, and in situ
u is a photograph showing the result of performing hybridization. A, autoradiogram, B, images of the same section stained with hematoxylin. Li: liver.

[Fig. 7] A section was prepared from a fetus at 14.5 days of fetal life, and in situ
u is a photograph showing the result of performing hybridization. A, C, autoradiogram, B, D, images of the same section stained with hematoxylin. Li: liver, Lu: lung, T
h: thymus, N: neural tube.

FIG. 8 shows a comparison of the amino acid sequences of human and mouse YS68.

FIG. 9 compares the amino acid structures of human (Human) and mouse (Mouse) YS68.

FIG. 10 is a photograph showing the result of analyzing a protein that co-precipitates with YS68. The liver was firstly cultured on embryonic day 14.5 to prepare a cell lysate. Anti-YS68 antibody and protein A
(Lane 1), rabbit IgG and protein A (lane 2), protein
Only A (lane 3) was immunoprecipitated. After SDS PAGE, the gel was stained with silver. Arrow; YS68, *: Protein co-precipitated with YS68.

FIG. 11 is a photograph showing the result of immunostaining of a YS68 tissue. Dorsal aort of mouse embryonic day 11.5 (dorsal aort)
a) (A, B, C, D, E), umbilical artery
(F), embryonic day 9 yolk sac blood vessels (H) were converted to erythroid marker TER
The cells were stained with 119 (A, B, G) and anti-YS68 antibody (C, D, E, H).
B and D show enlarged views of A and C, respectively, and E shows another view of the aorta. Arrows indicate where blood cells are sprouting from the vascular endothelium.

FIG. 12. Primary cultured cells of liver at 14.5 days of embryonic age were cultured with anti-YS68
FIG. 2 is a photograph showing the results of staining with an antibody (A) or rabbit IgG (B). YS68 was strongly expressed in and around the nucleus.

FIG. 13: Blood cells were separated from the liver on day 14 of embryonic embryos, and YS
14 is a photograph showing the result of examining expression of 68. Giemsa-stained liver blood cells (A), embryonic 14.5 day liver blood cells
(B), CD34 negative cells (C) and CD34 positive cells (D) were stained with anti-YS68 antibody. (EH) It was confirmed whether the actually sorted cells were CD34-positive. EF and GH have the same field of view, E and G indicate fluorescence imaging, and F and H indicate bright field imaging. Most of the cells sorted on CD34 were CD34 weak to strong positive (E, F). CD3
Cells that passed through the four columns hardly expressed CD34 (G, H).

FIG. 14 is a photograph showing the localization of YS68 in cells. The left photo shows a low magnification and the right photo shows a high magnification. Cells derived from fetal liver were stained with an anti-YS68 antibody to examine the endogenous YS68 expression site (upper row). In COS7 cells, pEFBOSE-F-YS68 (5-1148) expressing the N-terminal region of YS68 (middle) or pEFBOSE-F-YS68 (981-224) expressing the C-terminal region of YS68 were used.
3) was transfected (lower) and stained with an anti-Flag antibody to examine the intracellular localization.

FIG. 15A shows the region of YS68 used as bait. Numbers indicate amino acid numbers. These regions were expressed in adenine- and histidine-requiring yeast (AH109), and were wound on a plate lacking adenine and histidine and supplemented with X-?-Gal. As a result, it was revealed that a yeast strain expressing the 1080-1630 region grew on this plate, and that this region had a transcription activating ability. B is a photograph showing the result of detecting the transcription activation ability of each region of YS68.

FIG. 16 is a photograph showing a result of yeast two-hybrid screening. YS68 35-604 (upper), 583-1134
(Lower) AH109 strain expressing vector (1), protein
kinase C inhibitor (2), EST1 (3), EST2 (4), CD27
binding protein 1 (5), CD27 binding protein 1 (6)
And leucine, tryptophan-deficient (-LT) plate (left), leucine, tryptophan, histidine,
Adenine deletion, X-?-Gal added (-LTHA) plates were plated.

──────────────────────────────────────────────────の Continued on the front page (51) Int.Cl. ⁷ Identification symbol FI Theme coat ゛ (Reference) C12N 1/21 C12P 21/02 C 4H045 5/10 C12Q 1/68 A C12P 21/02 G01N 33/15 Z C12Q 1/68 33/50 Z G01N 33/15 33/53 D 33/50 C12P 21/08 33/53 C12N 15/00 ZNAA // C12P 21/08 5/00 A (31) Priority claim number Patent application 2000-123721 (P2000-123721) (32) Date of priority April 19, 2000 (April 19, 2000) (33) Countries claiming priority Japan (JP) Application for Patent Law Article 30 (1) has been filed August 25, 2011 Published in “Biochemistry, Vol. 71, No. 8” published by The Biochemical Society of Japan F term (reference) 2G045 AA34 AA35 AA40 DA12 DA13 DA36 FB01 FB02 FB03 FB05 FB06 4B024 AA01 BA41 BA80 CA04 CA09 CA11 DA02 DA12 EA04 GA03 GA13 HA01 HA13 4B063 QA01 QA18 QQ02 QQ08 QQ42 QQ52 QR08 QR32. FA72 FA74

Claims (14)

[Claims] 1. The DNA according to any one of the following (a) to (d). (A) DNA encoding a protein consisting of the amino acid sequence of SEQ ID NO: 12 or 14. (B) DNA containing the coding region of the nucleotide sequence of SEQ ID NO: 11 or 13. (C) an amino acid sequence represented by SEQ ID NO: 12 or 14 wherein one or more amino acids are substituted, deleted, inserted, and / or added; Encoding a protein functionally equivalent to the protein
A. (D) a DNA which hybridizes with a DNA consisting of the nucleotide sequence of SEQ ID NO: 11 or 13 under stringent conditions, and functionally binds to a protein consisting of the amino acid sequence of SEQ ID NO: 12 or 14 DNA encoding equivalent protein. 2. A DNA encoding a partial peptide of a protein consisting of the amino acid sequence of SEQ ID NO: 12 or 14. 3. A protein or peptide encoded by the DNA according to claim 1 or 2. 4. A vector into which the DNA according to claim 1 or 2 has been inserted. 5. A host cell carrying the vector according to claim 4. 6. The method for producing a protein or peptide according to claim 3, comprising a step of culturing the host cell according to claim 5, and recovering the expressed protein from the host cell or a culture supernatant thereof. 7. A polynucleotide comprising at least 15 nucleotides complementary to a DNA consisting of the nucleotide sequence of SEQ ID NO: 11 or 13 or a complementary strand thereof. 8. A method for screening a compound that binds to a protein according to claim 3, wherein (a) a step of bringing a test sample into contact with the protein or a partial peptide thereof, and (b) a protein or a partial peptide thereof. Detecting a binding activity between the protein and a test sample; and (c) selecting a compound having an activity of binding to the protein or a partial peptide thereof. 9. A compound that binds to the protein according to claim 3. 10. The compound according to claim 9, which is an antibody. 11. A compound that binds to the protein of claim 3, which can be isolated by the method of claim 8. 12. The protein according to claim 3 and PKCI or
A method for screening a compound that regulates binding to siva1, comprising: (a) contacting the protein or its partial peptide according to claim 3 with PKCI or siva1 in the presence of a test sample; Detecting the binding of
(C) selecting a compound having an activity of promoting or inhibiting the binding of the compound as compared to the case where the compound is detected in the absence of the test sample. 13. The protein according to claim 3 and PKCI or
A compound that regulates binding to siva1. 14. The protein of claim 3 and PKCI or siva, which can be isolated by the method of claim 13.
A compound that regulates binding to 1.