JPH08301898A - New polypeptide, its production, dna coding the polypeptide, vector comprising the dna, host cell transformed with the vector, antibody of the polypeptide, and pharmacological composition containing the peptide or antibody - Google Patents

Abstract

PURPOSE: To obtain a new polypeptide having a specific amino acid sequence, produced by the mallow stroma cells of a mouse, and useful for preventing and treating diseases relating to the aplasia or abnormal multiplication of hematopoietic cells, the sthenia or depression of nervous system functions and immune system functions, etc. CONSTITUTION: A new polypeptide having an amino acid sequence containing an amino acid sequence of the formula and comprising 361 amino acids produced from mouse mallow stroma cells, and a human homologue comprising 362, 254 and 450 amino acids, its fragment or the homologue of the fragment. The new polypeptide is useful for medicines for treating nerve damages or diseases related to the sthenia and depression of nervous system functions and immune system functions, etc., for medicines for preventing and treating bone metabolism disorders, etc. The polypeptide is obtained by extracting a mRNA from a mouse stroma cell strain SR2, producing a cDNA library in the presence of the mRNA as a template, screening the produced cDNA library, recovering a DNA from the positive clone, binding the DNA to a vector, and subsequently expressing the DNA in host cells.

Description

Detailed Description of the Invention

[0001]

The present invention relates to a novel polypeptide,
A method for producing the same, a DNA encoding the polypeptide,
It relates to a vector consisting of the DNA, a host cell transformed with the vector, an antibody of the polypeptide, and a pharmaceutical composition containing the peptide or antibody.
More specifically, a novel polypeptide produced by a certain mouse stromal cell line, a human homologue of the polypeptide, a method for producing the polypeptide, a DNA encoding the polypeptide, a vector comprising the DNA, The present invention relates to host cells transformed with the vectors, antibodies to the polypeptides, and pharmaceutical compositions containing the polypeptides or antibodies.

[0002]

BACKGROUND OF THE INVENTION Bone marrow stromal cells form the bone marrow microenvironment of the immune system, hematopoietic system, etc., and are factors that are essential for inducing proliferation and differentiation of these stem cells, such as IL-7, SCF, I.
L-11, M-CSF, G-CSF, GM-CSF, I
It is known to produce and secrete factors such as L-6, TGF-β and LIF. It has also been revealed that some bone marrow stromal cells are involved in bone metabolism.
However, with only the isolated factor groups so far,
It cannot fully play the role of stromal cells. This suggests that there are factors that have not yet been isolated.

[0003]

OBJECTS OF THE INVENTION The present inventors have paid attention to this point, and in order to find out the novel factor (polypeptide) produced by a certain type of stromal cell, particularly secretory protein and membrane protein, Diligent study was conducted.

Conventionally, in order to obtain a specific polypeptide or a DNA encoding the same, a desired biological activity was confirmed in a tissue or cell culture medium, and then the polypeptide was isolated and purified to obtain a gene. , Or a method of expressing and cloning a gene using its biological activity as an index has been generally used. However, in vivo biologically active polypeptides often have various biological activities, and as a result of cloning a gene using a certain activity as an index, it was confirmed that it was identical to a known polypeptide. Increasing numbers of cases are being discovered later. Moreover, most of the factors produced by bone marrow stromal cells are produced only in a small amount, which makes isolation, purification and confirmation of biological activity difficult.

[0005] In recent years, cDNA production technology and sequencing technology have rapidly developed, and a large amount of cDNA can be sequenced rapidly. Therefore, using these techniques, cDNA libraries were prepared from various cells and tissues, cDNA was randomly cloned to determine the nucleotide sequence, the corresponding polypeptide was expressed, and the physiological function was analyzed. The method of doing is developing. This method has the feature that a gene can be cloned and the information of its nucleotide sequence can be obtained without requiring any biochemical or genetic analysis. Large contingent element.

The present inventors have so far studied cloning of genes for growth differentiation factors that work in the hematopoietic system and the immune system. Most of secretory proteins such as growth and differentiation factors (for example, various cytokines) and membrane proteins such as their receptors (hereinafter collectively referred to as secretory proteins and the like) have a signal peptide at their N-terminals. Focusing on the fact that it has a so-called sequence, we have intensively studied a method for efficiently and selectively cloning a gene encoding a signal peptide. As a result, we found a method for efficiently amplifying the N-terminal fragment and easily searching for the presence or absence of a signal peptide (see Japanese Patent Application No. 613951), and using this method, bone marrow stromal cells are produced. The present invention has been completed by succeeding in finding a novel factor (peptide) and DNA encoding the same.

Swiss Plot (Swiss Prot Release 29
As a result of investigating the amino acid sequence of the known polypeptide registered in (4), the polypeptide of the present invention has a calcium binding protein (Ca binding protei) such as Calmodulin and Troponin C.
n) EF-hand (calcium binding region (Ca bindin
g domein)) motif, but no homologous or similar sequence was found. From this, it was confirmed that the polypeptide of the present invention is a novel calcium-binding protein.

[0008]

The present invention comprises (1) a polypeptide comprising the amino acid sequence shown by SEQ ID NO: 1, (2) the above (1)
DNA encoding the polypeptide described in (3), DNA having the nucleotide sequence shown in SEQ ID NO: 2, (4) DNA having the nucleotide sequence shown in SEQ ID NO: 3, (5) SEQ ID NO: 5, 9 or 13 A polypeptide comprising the amino acid sequence represented by: (6) a DNA encoding the polypeptide described in (5) above, (7) a DNA having the nucleotide sequence represented by SEQ ID NO: 6, 10 or 14, and (8) It relates to a DNA having the nucleotide sequence shown in SEQ ID NO: 7, 11 or 15.

The present invention relates to a polypeptide consisting of the amino acid sequence shown in SEQ ID NO: 1, 5, 9 or 13 in a substantially pure form, a homologue thereof, a fragment of the sequence and a homologue thereof. The invention further relates to DNA encoding those polypeptides. More specifically, SEQ ID NOs: 2, 3, 6, 7, 10, 11, 1
DNA having the nucleotide sequence shown by 4 or 15, and SEQ ID NO: 2, 3, 6, 7, 10, 11, 14 or 1
The present invention relates to a DNA having a fragment that selectively hybridizes to the base sequence shown in 5.

SEQ ID NOs: 1, 5 in substantially pure form
The polypeptide having the amino acid sequence represented by 9 or 13 is generally a polypeptide having 90% or more, for example, 95, 98 or 99% of the polypeptide at the time of production, having the amino acid sequence represented by SEQ ID NO: 1. Means that.

The homologue of the polypeptide consisting of the amino acid sequence shown in SEQ ID NO: 1, 5, 9 or 13 is
Generally at least 20, preferably at least 30
At least 70%, preferably at least 8 in a contiguous amino acid region of 40, 60 or 100 amino acids, for example
The homologues are 0 or 90%, more preferably 95% or more homologous, and such homologues are hereinafter described as the polypeptide of the present invention.

Further, a fragment of the polypeptide consisting of the amino acid sequence shown in SEQ ID NO: 1, 5, 9 or 13 or a homologue fragment thereof is
At least 10 amino acids, preferably at least 15 amino acids, such as 20, 25, 30, 40, 50 or 6
It means 0 amino acid moiety.

SEQ ID NOs: 2, 3, 6, 7, 10, 11, 1
The DNA which selectively hybridizes to the DNA having the nucleotide sequence represented by 4 or 15 is generally at least 20, preferably at least 30, for example 40,
At least 70%, preferably at least 80 or 90% in 60 or 100 contiguous base sequence regions,
More preferably, it is 95% or more homologous, and such a DNA is hereinafter described as the DNA of the present invention.

SEQ ID NOs: 2, 3, 6, 7, 10, 11, 1
The fragment of DNA having the nucleotide sequence represented by 4 or 15 means at least 10 bases, preferably at least 15 bases, for example, 20, 25, 30 or 40 base portions, and such a fragment is also DN of the present invention.
Included in A.

Further, the present invention includes a replication or expression vector comprising the DNA of the present invention. Examples of the vector include a plasmid, virus or phage vector comprising an ori region and, if necessary, a promoter for expressing the above-mentioned DNA, a promoter regulatory factor and the like. The vector may include one or more selectable marker genes, such as the ampicillin resistance gene. The vector is in vit
ro), for example, production of RNA corresponding to DNA,
It can be used to transform host cells.

Further, in the present invention, SEQ ID NOs: 2, 3 and
Also included is a host cell transformed with a vector for replicating or expressing the DNA of the present invention containing the DNA having the nucleotide sequence of 6, 7, 10, 11, 14 or 15 or an open reading frame thereof. . Cells include, for example, bacteria, yeast, insect cells or mammalian cells.

Further, the present invention also includes a method for producing the polypeptide of the present invention, which comprises culturing the host cell of the present invention under conditions for expressing the polypeptide of the present invention. Culturing is preferably carried out under conditions in which the polypeptide of the present invention is expressed and produced by a host cell.

The DNA of the present invention is inserted into the antisense region of the vector as described above to obtain antisense RN.
A can also be produced. Such antisense R
NA can also be used to control the levels of the polypeptides of the invention in cells.

The present invention also includes monoclonal or polyclonal antibodies of the polypeptides of the present invention. Furthermore, the method for producing a monoclonal or polyclonal antibody of the polypeptide of the present invention is also included. A monoclonal antibody can be produced by a usual hybridoma technique using the peptide of the present invention or a fragment thereof as an antigen. The polyclonal antibody can be produced by a usual method in which a host animal (for example, rat or rabbit) is inoculated with the polypeptide of the present invention and immune serum is collected.

The present invention also includes a pharmaceutical composition containing the polypeptide of the present invention, an antibody thereof and a pharmaceutically acceptable excipient and / or carrier.

The polypeptide of the present invention (1) (SEQ ID NO: 1) or (5) (SEQ ID NO: 5, 9 or 13) has the amino acid sequence shown in SEQ ID NO: 1 or 5, 9 or 13. In addition to the above, those lacking a part thereof (for example, in SEQ ID NO: 1 or 5, 9 or 13 lacking a polypeptide consisting of only an essential part for expression of biological activity, cytoplasmic domain, transmembrane domain region) Soluble polypeptides, etc.), those in which a part thereof is replaced with other amino acids (for example, those in which an amino acid having similar physical properties are substituted), and those in which other amino acids are added or inserted are also included. As is well known, the codon that encodes one amino acid is 1 to
Six types (for example, one type for Met and six types for Leu) are known. Therefore, the base sequence of DNA can be changed without changing the amino acid sequence of the polypeptide.

The DNA of the present invention specified in (2) or (6) includes SEQ ID NO: 1 or (5) of each (1).
All base sequence groups that encode the polypeptide shown in SEQ ID NO: 5, 9 or 13 are included. The productivity of the polypeptide may be improved by changing the base sequence. (3) (SEQ ID NO: 2) or (7) (SEQ ID NO: 6, 10)
Alternatively, the DNA specified in 14) is one embodiment of the DNA shown in each of (2) or (6) and represents a native sequence. (4) (SEQ ID NO: 3) or (8) (SEQ ID NO: 7, 11)
Alternatively, the DNA shown in 15) shows the sequence obtained by adding the natural untranslated portion to the DNA specified in (3) or (7).

D having the nucleotide sequence shown in SEQ ID NO: 3
The NA is produced according to the following method.

Preparation of cDNA Library of Signal Peptide (1) After synthesizing single-stranded DNA for mRNA isolated from target cells using random primers, 3'end of the obtained single-stranded DNA (2) A double-stranded DNA for the single-stranded DNA obtained in (1) was synthesized using a poly C oligomer linked with a specific restriction enzyme (enzyme I) site as a primer. , (3) disrupt the double-stranded DNA obtained in (2),
After fractionation by the fragment size, a linker containing a specific restriction enzyme (enzyme II) site different from enzyme I is ligated and fractionated again (4) A primer containing enzyme I site and a primer containing enzyme II site CD amplified by polymerase chain reaction (hereinafter abbreviated as PCR) using
NA was digested with enzyme I-enzyme II and then fractionated, and (5) the obtained cDNA fragment was ligated upstream of a gene such as a known secretory protein from which a signal peptide was deleted to obtain a eukaryotic expression plasmid vector. It comprises a step of transforming after being incorporated.

Each step will be described in detail. In step (1), the cells of interest are stimulated with an appropriate stimulant if necessary, and then the method (Method i) such as Okayama H.
n Enzymology, 154 , 3 (1987)).
Isolation of A is performed. The target cells may be any cells that may possibly produce secretory proteins and the like. Examples include neural cells and hematopoietic cells.
The synthesis of single-stranded cDNA using random primers is performed by a known method. A commercially available random primer is used. Subsequently, oligo dC is added to the 3'end of the single-stranded cDNA by terminal deoxytransferase.

The double-stranded DNA in step (2) is also synthesized by a known method. Any restriction enzyme (enzyme I) site linked to the poly G oligomer serving as a primer and a restriction enzyme (enzyme II) site used in the next step (3) may be used as long as they are different from each other. Preferably, Sal I is used as enzyme I and Sac I is used as enzyme II.

In step (3), the cDNA is processed by ultrasonication.
Fragmented to an average length of 500 bp and 400-800 bp cDNA by agarose electrophoresis (AGE).
After fractionation into A, the ends are blunted with T4 DNA polymerase, an enzyme II adapter is ligated, and again fractionated into a DNA of 400 to 800 bp by agarose electrophoresis. The enzyme II may be any enzyme different from the enzyme I as described above. This step increases the possibility that the cDNA fragment containing the signal peptide will be present in the region sandwiched by enzyme I and enzyme II.

In step (4), PCR is carried out in order to further enhance the establishment of the portion sandwiched between enzyme I and enzyme II which may contain a signal peptide. PCR is a well-known method, and automated devices therefor are commercially available. Amplification of 25 to 30 times is sufficient. Amplified cd
NA is digested with enzyme I-enzyme II and fractionated into cDNA of 400 to 800 bp by agarose electrophoresis.

In the step (5), a gene such as a known secretory protein in which a signal peptide has been deleted from a plasmid vector for eukaryotic cell expression (referred to as a reporter gene) and the cDNA fragment obtained in (4) upstream thereof. Is a step of transforming by incorporating. Although various types of eukaryotic expression plasmid vectors are known here, for example, pcDL-SRα and pcEV- that function in E. coli.
4 is used.

Further, as the reporter gene, genes of soluble secretory proteins of all kinds and mature protein portions of membrane proteins are used. Furthermore, the expression of these reporter genes must be confirmed by some method such as antibody method. The human IL-2 receptor α gene is preferably used. Many host Escherichia coli strains for transformation are already known, and any one may be used, but DH5 competent cells (described in Gene, 96 , 23 (1990)) are preferable.
The transformant is cultured by a conventional method to obtain the cDNA library of the present invention (a conceptual diagram is shown in FIG. 1).

In the method for preparing a cDNA library of the present invention, it is highly possible that the library contains a gene fragment encoding a signal peptide, but not all clones contain the fragment, and all are unknown. It is not always the gene fragment encoding the (novel) signal peptide of. Therefore, next, it is necessary to screen a gene fragment encoding an unknown signal peptide from the library.

That is, the cDNA library is subdivided into pools having an appropriate size and incorporated into an expression system. Expression systems for producing a polypeptide include mammalian cells (eg, monkey COS-7 cells, Chinese hamster CHO cells, mouse L cells, etc.). Transfection is performed by a known method, for example, the DEAE-dextran method. After culturing, the presence or absence of expression of the reporter gene is determined.

The reporter gene is known to be expressed even if the signal peptide is unique to other secreted proteins. That is, expression of the reporter gene indicates that a signal peptide of some secretory protein was incorporated in the library. The positive pool is further subdivided, and expression and determination are repeated until a single clone is obtained. The determination of the expression of the reporter gene depends on the type of the reporter gene.
Law) etc.

Next, the nucleotide sequence of the isolated positive clone was determined, and for the cDNA that was found to encode an unknown protein, the full-length clone was isolated by using it as a probe to obtain the full-length nucleotide sequence. You can decide. These operations are all performed by methods known to those skilled in the art. For example, the nucleotide sequence is determined by the Maxam-Gilbert method or the dideoxy terminator method. The full-length sequence is based on Molecular Cloning (Sambrook, J., Frits
ch, EF and Maniatis, T., Cold Spring Harb
or Laboratory Press, published in 1989).

When the nucleotide sequences shown in SEQ ID NOS: 2 and 3 are partially, preferably all, determined, the DNA encoding the protein of the present invention present in mammals or the DNA encoding homologues and subsets of the protein of the present invention. Can be obtained. By synthesizing an oligonucleotide having an appropriate mouse nucleotide sequence and using it, a cDNA library or mRNA derived from a mammal is subjected to a PCR method, or by hybridizing a fragment of the appropriate mouse nucleotide sequence as a probe,
DNA encoding a mammalian type protein of interest from a mammalian cDNA library or genomic library
Can be obtained.

More specifically, for example, (i) in order to isolate the cDNA encoding the human peptide of the present invention, cells producing the peptide can be selected by Northern blot analysis or PCR. (ii) It has been confirmed by Northern blotting or the like that a gene for a human homologue of the mouse peptide has been expressed, for example, mRNA is isolated from a cell line or tissue, and (iii) the first strand (single-stranded cDNA is isolated from the mRNA. ), Then a second strand (double-stranded cDNA) was synthesized (cDN
Synthesis of A), (iv) incorporating the cDNA into an appropriate phage vector, (v) infecting a host cell with the obtained recombinant phage (preparation of cDNA library), (vi)
Screening the obtained cDNA library by plaque hybridization using a cDNA fragment encoding the labeled peptide as a probe,
(vii) Phage DNA from the obtained positive clone
Can be prepared by subcloning the excised cDNA into a plasmid vector and sequencing (viii) full length.

More specifically, step (i) comprises the step of stimulating a human glioblastoma cell line with a suitable stimulant (eg IL-).
(1 etc.) or left unstimulated, Okayama et al. (Method in Enzymology, 154 , 3 (1987))
According to. The cell producing the present polypeptide is preferably the human glioblastoma cell line T98.
G (ATCC strain number, CRL-1690),
Any cell may be used as long as it produces the peptide.

The process of the process (ii) is Okayama (H)
Et al. (See Method in Enzymology, 154 , 3 (1987))
According to. The steps (iii), (iv) and (v) are steps for preparing a cDNA library and are performed according to the modified Gubler & Hoffman method (described in Gene, 25 , 263 (1983)).

The plasmid vector used in step (iv) is a plasmid vector (for example, pBR32
2, pBluescript II, etc.) and phage vectors (eg,
λgt10, λDASH II, etc.) are known, but preferably the phage vector λgt22A (42.73k).
bp sold by BRL) is used.

The host cells used in step (v) include
Preferably, Escherichia coli Y1090 (r-) (sold by BRL)
Is used. Steps (vi) and (vii) are based on Molecular Cl
oning (Sambrook, J, Fritsh, E, F. and Maniatis,
T, Cold Spring Harbor Laboratory Press (1989)). Sequencing of step (viii) is performed by Maxam-Gilbert
Method and dideoxy terminator method.

The cDNA thus obtained is the c
It is necessary to confirm that the DNA is full length or almost full length. This confirmation uses the cDNA as a probe,
Northern analysis (see Mole above)
See cular Cloning). The size of the mRNA obtained from the hybridized band and the size of the cDNA are compared, and if they are almost the same, the cDNA is considered to be almost full length.

SEQ ID NOs: 2, 3, 6, 7, 10, 11, 1
Once the base sequences shown in 4 and 15 are confirmed,
Thereafter, the DNA of the present invention can be obtained by chemical synthesis, or by chemically synthesizing a fragment of the base sequence and hybridizing it with a probe. Furthermore, the target DNA can be obtained in a required amount by introducing the vector DNA containing the present DNA into an appropriate host and allowing it to grow.

As the method for obtaining the polypeptide of the present invention, (1) a method of purifying and isolating from living organisms or cultured cells, (2) a method of peptide synthesis, or (3) a method of producing using a gene recombination technique. , And the like, but the method described in (3) is industrially preferable.

Examples of the expression system (host-vector system) for producing a polypeptide using gene recombination technology include expression systems of bacteria, yeast, insect cells and mammalian cells.

For example, in the case of expression in Escherichia coli, a start codon (ATG) is added to the 5'end of the DNA encoding the mature protein portion, and the resulting DNA is used as an appropriate promoter (for example, trp promoter, lac). (Promoter, λPL promoter, T7 promoter, etc.)
And is inserted into a vector that functions in E. coli (for example, pBR322, pUC18, pUC19, etc.) to prepare an expression vector.

Next, E. coli transformed with this expression vector (for example, E. Coli DH1, E. Coli JM109,
E. Coli HB101 strain) can be cultured in an appropriate medium to obtain the target polypeptide from the cells. Also, bacterial signal peptides (eg p
By using the signal peptide of elB), the desired polypeptide can be secreted into the periplasm. Furthermore, it is also possible to produce a fusion protein with another polypeptide.

When expressed in mammalian cells, for example, a DNA having the nucleotide sequence represented by SEQ ID NO: 3 or 7 is used as a suitable vector (eg, retrovirus vector, papillomavirus vector, vaccinia virus vector, SV40). Appropriate promoter (eg, SV40 promoter, LTR)
(A promoter, a metallothionein promoter, etc.) to prepare an expression vector. Then, using the obtained expression vector, suitable mammalian cells (for example, monkey CO
S-7 cells, Chinese hamster CHO cells, mouse L cells, etc.) and the transformant is cultured in an appropriate medium, whereby the desired polypeptide is secreted into the culture solution. The polypeptide obtained as described above can be isolated and purified by a general biochemical method.

[0048]

The polypeptide of the present invention (human type α, β,
γ and mouse α are calmodulin (Calmoduli)
n) and homology with troponin C and other calcium binding proteins. These known proteins include those around 150aa (including EF-hand motief) or 25 of the polypeptide of the present invention.
Around 0aa (including EF-hand-like sequences), α type 300a
Homology was observed with at least one of the three partial sequences (20-50 aa length) near a (including EF-hand motief). In addition, EF-hand (Ca bin
ding domein; 109-122aa, 148-161aa, 289-302aa). From this, the polypeptide of the present invention is a protein that functions under the control of Ca, or Ca
It is considered to be a protein involved in the preparation of 3 types of c
DNA is an alternative splicing (Alternativ
esplicing), which results in different amino acids at the C-termini, so it is possible that the region is changed. Furthermore, since the polypeptide of the present invention is produced and secreted by a stromal cell line, differentiation, proliferation, growth or survival maintenance of hematopoietic cells, biological activity associated with the function of the immune system, It is considered to have biological activity related to tumor growth, growth or inflammation, and biological activity related to bone metabolism.

For example, proliferation of B cells, T cells and mast cells, class-specific induction by promoting immunoglobulin class switching, differentiation of B cells into antibody-producing cells, proliferation or differentiation of granulocyte progenitor cells, monocytes / Proliferation or differentiation of macrophage progenitor cells, neutrophils, monocytes / macrophages,
Eosinophil, basophil proliferation or hyperactivity, megakaryocyte progenitor cell proliferation, neutrophil progenitor cell proliferation or differentiation, B or T progenitor cell proliferation or differentiation, erythrocyte production promotion, erythrocyte, neutrophil , Eosinophil, basophil, monocyte / macrophage, mast cell, megakaryocyte progenitor cell growth support, neutrophil, monocyte / macrophage, B cell or T cell migration promotion,
Thymocyte proliferation, adipocyte differentiation inhibition, natural killer cell proliferation, hematopoietic stem cell proliferation, stem cell and various hematopoietic progenitor cell proliferation inhibition, or promotion of bone resorption by promoting monocyte to osteoclast differentiation, There is a possibility that it may have an action of promoting the differentiation or proliferation of leaf-like stem cells to osteoblasts or of activating osteoclasts by the present polypeptide alone or by acting synergistically with other cytokines.

In addition to these, it is predicted that any factor that acts on the immune system will also act on the nervous system. Therefore, differentiation into various neurotransmitter-acting nerve cells and their survival maintenance, glial It may also act to promote cell proliferation, neurite outgrowth, ganglion cell survival maintenance, astrocyte proliferation or differentiation promotion, peripheral nerve proliferation or survival maintenance, Schwann cell proliferation, motor nerve proliferation or survival maintenance There is a nature.

Furthermore, the present polypeptide is used in the process of early embryonic development to form organs of the epidermis, brain, spine, and nerves by the action of ectoderm, and connective tissues of the dorsal cord (bone, muscle, tendon), blood cells by the action of mesoderm induction. Promotes or suppresses the formation of cells, heart, kidneys, gonads, or the formation of digestive organs (stomach, intestine, liver, pancreas) and respiratory system (lung, trachea) by endoderm induction In addition to the above, there is a possibility that the above-mentioned organs also have a growth or growth-suppressing action on the above-mentioned organs.

Therefore, the polypeptide of the present invention is, by itself, a disease associated with poor development or abnormal proliferation of hematopoietic cells, a decrease or enhancement of the function of the immune system or nervous system or bone metabolism, such as an inflammatory disease (rheumatic, Ulcerative colitis, etc.), hematopoietic stem cell depletion after bone marrow transplantation, leukocytes after irradiation or chemotherapy for cancer and leukemia,
Platelet, B cell or T cell depletion, anemia, infection,
Remedies for cancer, leukemia, AIDS, various degenerative diseases (Alzheimer's disease, multiple sclerosis, etc.) or nerve damage,
It is expected to be used as a preventive or therapeutic drug for abnormal bone metabolism (osteoporosis, etc.).

Further, since the present polypeptide may have the action of differentiating or proliferating organs derived from ectoderm, mesoderm or endoderm, each polypeptide (epidermis, bone, muscle, tendon, heart, kidney, stomach, intestine) , Liver, pancreas, lung, trachea, etc.).

Further, the amount of the polypeptide in the living body can be quantified by using the polyclonal antibody or the monoclonal antibody of the polypeptide, and thus it can be used for the study of the relationship between the polypeptide and the disease or the diagnosis of the disease. it can. Polyclonal antibodies and monoclonal antibodies can be prepared by a conventional method using the polypeptide or a fragment thereof as an antigen.

The DNA of the present invention not only serves as an important and essential template for producing the polypeptide of the present invention, which is expected to have great utility, but also can be used for diagnosis and treatment of genetic diseases (treatment of gene deficiency). Alternatively, it can be used for the treatment by stopping the expression of the polypeptide by antisense DNA (RNA). Moreover, genomic DNA can be separated using the DNA of the present invention as a probe. Similarly, it is also possible to isolate a gene of a human related polypeptide having high homology with the DNA of the present invention or a gene of a polypeptide highly homologous to the polypeptide of the present invention in an organism other than human.

[0056]

[Application to pharmaceuticals] Developmental failure or abnormal proliferation of hematopoietic cells,
Diseases associated with increased or decreased nervous system function, or increased or decreased immune system function, such as inflammatory diseases (rheumatism, ulcerative colitis, etc.), hematopoietic stem cell depletion after bone marrow transplantation, post-radiotherapy or chemotherapy administration Subsequent white blood cells, platelets, B or T cell depletion, anemia, infection, cancer, leukemia, AI
The polypeptide of the present invention for the prevention or treatment of DS, various degenerative diseases (Alzheimer's disease, multiple sclerosis, etc.) or nerve damage, the prevention or treatment of abnormal bone metabolism (osteoporosis, etc.), tissue repair, etc. Is usually administered systemically or locally, generally in oral or parenteral form. Oral administration, intravenous administration and intracerebroventricular administration are preferred.

The dose depends on age, body weight, symptoms, therapeutic effect,
Depending on the administration method, treatment time, etc., it is usually administered once per adult in the range of 100 μg to 100 mg once or several times a day orally, or 10 μg per adult per administration. In the range of 1 to 100 mg, it is parenterally administered once to several times a day. Of course, as described above, since the dose varies depending on various conditions, a dose smaller than the above dose may be sufficient in some cases, or a dose exceeding the range may be necessary.

When the compound of the present invention is administered, it is used as solid compositions, liquid compositions and other compositions for oral administration, injections for parenteral administration, external preparations, suppositories and the like. Solid compositions for oral administration include tablets,
Pills, capsules, powders, granules and the like are included. Capsules include soft capsules and hard capsules.

In such solid compositions, the one or more active substances comprise at least one inert diluent (eg lactose, mannitol, glucose, hydroxypropylcellulose, microcrystalline cellulose, starch, polyvinyl). Pyrrolidone, magnesium aluminometasilicate, etc.). The composition may be an additive other than an inert diluent, such as a lubricant (magnesium stearate, etc.), a disintegrating agent (calcium fibrin glycolate, etc.), a stabilizer (human serum albumin, lactose, etc.) according to a conventional method. ), And a solubilizing agent (arginine, aspartic acid, etc.) may be contained.

The tablets or pills may be coated with a gastric or enteric film of sucrose, gelatin, hydroxypropylcellulose, hydroxypropylmethylcellulose phthalate or the like, if necessary, or with two or more layers. Good. Also included are capsules of absorbable material such as gelatin.

Liquid compositions for oral administration include pharmaceutically acceptable emulsions, solutions, suspensions, syrups, elixirs and the like, and generally used inert diluents (eg, Purified water, ethanol, etc.) may be contained. Such a composition comprises a wetting agent, in addition to an inert diluent,
An auxiliary agent such as a suspension agent, a sweetening agent, a flavoring agent, an aromatic agent, and a preservative may be contained.

Other compositions for oral administration include spray formulations which contain one or more active substances and are formulated in a manner known per se. This composition contains, in addition to an inert diluent, a stabilizer such as sodium bisulfite and a stabilizer that imparts isotonicity, an isotonic agent such as sodium chloride, sodium citrate or citric acid. May be. The method for producing the spray agent is described in, for example, U.S. Pat.
It is described in detail in No. 355.

The injections for parenteral administration according to the present invention include sterile aqueous or non-aqueous solutions, suspensions and emulsions. As aqueous or non-aqueous solutions or suspensions, one or more active substances are mixed with at least one inert diluent. Examples of the aqueous diluent include distilled water for injection and physiological saline. Examples of non-aqueous diluents include propylene glycol, polyethylene glycol, vegetable oils such as olive oil, alcohols such as ethanol, and Polysorbate 80 (registered trademark).

Such a composition may further contain a preservative, a wetting agent, an emulsifying agent, a dispersing agent, a stabilizing agent (eg, human serum albumin, lactose, etc.), a solubilizing agent (eg, arginine, aspartic acid, etc.). The auxiliary agent may be included.

These are sterilized by filtration through a bacteria-retaining filter, blending of a bactericide or irradiation.
They can also be used by preparing a sterile solid composition (for example, by a freeze-drying method) and dissolving it in sterile distilled water for injection or other solvent before use.

Other compositions for parenteral administration include external preparations, ointments, coatings, suppositories and pessaries for rectal administration, which contain one or more active substances and are formulated by a conventional method. Etc. are included.

[0067]

The present invention will be described in more detail with reference to the following examples and reference examples, but these do not limit the scope of the present invention.

Example 1: Construction of plasmid for producing expression vector The pcD-SRα296 vector produced by Takebe et al. (Mol.
Cell. Biol., 8 , 966 (1988)), SV40 early promoter, RTR R region of HTLV-I and U5.
Promoter system (SR
It is an excellent expression vector for mammalian cells having α). However, there are drawbacks such as (1) the cloning site of the insert is only EcoRI, and (2) the vector recovery from E. coli is low because the pBR322 vector is the backbone of the vector region. Therefore, pcD- which has a multi-cloning site of the insert with the pUC19 vector, which has a large yield from E. coli, as the backbone
The SRα296 modified vector was prepared by the following method.

The pcD-SRα296 vector (provided by Mr. Takebe, National Institute of Preventive Health) was cleaved with SalI, and the 1.7 kbp fragment containing the SRα promoter was separated and recovered by agarose electrophoresis. This fragment was treated with Klenow to make a blunt end. pU
2.4 kbp containing the region of AmpR and pUCori after cutting the C19 vector with Nde I and Hind III
Fragment was separated and collected by agarose gel electrophoresis, treated with Klenow to make blunt ends, and then B
It was treated with AP (bacterial alkaline phosphatase) to remove the phosphate group at the 5'end.

The 1.7 kbp fragment containing the SRα promoter and the 2.4 kbp fragment containing pUCori thus obtained were circularized by ligation to construct a new vector. A PstI-KpnI fragment was removed from the obtained vector, and a synthetic polylinker having the following T7 and sp6 promoters,

[0071]

Embedded image

Was replaced with The plasmid vector constructed in this way (about 3.9 kbp, shown in FIG. 2) was transformed into p.
It was named UCSRαML2.

PUCSRαML2 has the following characteristics as a multipurpose plasmid vector. 1. Okayama-Berg method and Gubler & Hoffman method can be applied. 2. High plasmid yield per culture. 3. Single-stranded DNA can also be prepared. 4. It is easy to cut out the cDNA insert. 5. Delation Mutant for Sequence
(Deletion mutant) can be easily produced. 6. Transcription in vitro is possible. 7. It has a promoter that is expressed in mammalian cells.

Example 2 Construction of Vector for Generating cDNA Library Selective for Signal Peptide Next, a protein in which the signal sequence of hTac (human IL-2 receptor α, used as a reporter gene) was removed was encoded. To the pUCSRαM
A plasmid incorporated into L2 was constructed, and pUCSRα-
It was named hTac. Incorporating a cDNA downstream of the SRα promoter of the vector and upstream of the hTac gene,
If a fusion protein is formed with hTac by translating a protein from a cDNA having a signal sequence,
The fusion protein will be expressed on the membrane.

(1) That is, pBluescript SK (+)
(Sold by Stratagene, pBS) pBS-hTac in which the hTac cDNA was incorporated into the Hind III site.
Was digested with KpnI, blunted with T4 DNA polymerase, then digested with SacI to remove the leader sequence and then ligated with the SacI-EcoRI adapter to obtain an EcoRI-blunted end fragment. S this fragment
EcoRI of pUCSRαML2 that destroyed the acI site
-Incorporate into the SmaI site, pUCSRαML2-h
Tac was obtained (shown in FIG. 3).

Example 3 Preparation of cDNA Library Selective for Signal Peptide Mouse stromal cell line ST2 (cells that support the survival and proliferation of hematopoietic stem cells and the proliferation and differentiation of B cells and myeloid cell lines, EMBO J., 7 , 1337 (1988)), AGPC (acid guanidine-phenol-chloroform) method (Cell engineering experiment protocol (published by Shujunsha), page 28-31. The total RNA was extracted with
Poly A-RNA was purified using T) -latex (Oligotex-dT30 (trade name, sold by Takara Shuzo Co., Ltd.)). Single-stranded cDNA was synthesized by reverse transcriptase using random hexamers as primers, and dC was added to its 3'end by terminal deoxytransferase. 17me in which a restriction enzyme site containing SalI was ligated
dC of r,

[0077]

Embedded image

Was annealed, and double-stranded cDNA was synthesized using it as a primer. After sonicating so that the average length was 500 bp to fragment the cDNA, 400-800 bp cDNA was fractionated by agarose gel electrophoresis. After blunting the ends with T4 DNA polymerase,
A lawn linker containing a SacI site,

[0079]

Embedded image

(See Nucreic Acids Res., 18 , 4293 (1990)) and ligated again by agarose electrophoresis.
The cDNA of 00 to 800 bp was fractionated. A primer containing a SalI site (NLC),

[0081]

[Chemical 4]

And a primer containing SacI site (LL
HES),

[0083]

Embedded image

PCR was performed for 25 cycles under conditions of 94 ° C. for 1 minute, 50 ° C. for 2 minutes, and 72 ° C. for 2 minutes. The amplified cDNA was digested with SacI and EcoRI, and then subjected to agarose gel electrophoresis at 400-800b.
The cDNA of p was fractionated. This cDNA and pUCSR
αML2-hTac (prepared in Example 2) was Sac.
I and the SalI-digested plasmid were ligated with T4 DNA ligase, and Escherichia coli DH5α strain was transformed to obtain a cDNA library having selectivity for the signal peptide.

Example 4 Screening and Analysis of cDNA Encoding Signal Peptide The colonies obtained from the library prepared in Example 3 were subdivided into pools (about 50 colonies / pool). For each pool, the plasmid was isolated by the miniplet method, and
EAE-Dextran method (Current Protocol in Molecu
lar Biology, §9.2.1) was used to transfect COS-7 cells. After 48 hours, cells were detached from the dish and incubated with mouse anti-Tac IgG antibody for 20 minutes on ice. After removing the free antibody, it was incubated with FITC (fluorescein isothiocyanate) -labeled goat anti-mouse IgG antibody for 20 minutes on ice to remove the free antibody again.
Fluorescent staining for Tac on the cell surface was performed. A positive pool was then selected using a fluorescence microscope. For one pool, the colonies were further subdivided, and the same method was repeated until a single clone was obtained.
One positive clone (pBS-ST-NT3) was obtained.
Next, two kinds of synthetic primers specific to the pUCSRαML2-hTac vector

[0086]

[Chemical 6]

The base sequence of the ST-NT3 insert was determined using. DNA sequencing was performed by the cycle sequencing method using the fluorescent dye-terminator of ABI (Applied Biosystems Inc.) based on the dideoxy terminator method of Sanger (F) et al. Also, for reading the sequence,
A DNA sequencer (Model 373A) manufactured by ABI was used. As a result of conducting a homology search with a database at the DNA and amino acid level, it was revealed that ST-NT3 encodes an unknown protein.

Example 5: Screening of full-length cDNA and determination of nucleotide sequence A cDNA library was prepared by using Superscript (Supe) from mRNA derived from mouse stroma cell line ST2.
r Script, a registered trademark) Lambda System (sold by BRL). This cDNA was ligated with λgt22A (sold by BRL) having phosphatase-treated SalI and NotI arms (arm). In vitro packaging is based on in vitro packaging
In vitro Packaging Kit According to the protocol of LAMDA INN (Nippon Gene), the recombinant phage was infected with host Escherichia coli Y1090 (r-) (sold by BRL). As a result, a cDNA library consisting of 1 million plaques was obtained. Next, pBS-S
T-NT3 was digested with SalI and NotI, and an ST-NT3 fragment was prepared by agarose electrophoresis. Oligo labeled S
A large number of positive clones were obtained by screening the library using the T-NT3 cDNA fragment as a probe. Among them, a clone with an insert of about 1.0 kbp was SalI-excised from the λgt22A vector.
The NotI fragment was subcloned into pUCSRαML2 to obtain plasmid pUCSRαML2-ST-NT3. Using the T7 primer, the nucleotide sequence of 300 bp on the 5'side of the ST-NT3 cDNA was determined, and the S-probe
First, it was confirmed that a sequence corresponding to T-NT3 was present at the most 5'side of ST-NT3 derived from the phage library.

Example 6: Determination of full-length sequence of cDNA and determination of open reading frame The full-length sequence of cDNA was Molecular Cloning.
(Sambrook, J., Fritsch, EF and Maniatis, T.
Random sequencing was performed according to the method described in "Cold Spring Harbor Laboratory Press" (published in 1989).

That is, the plasmid was recovered from the ST-NT3 clone, and the cDNA insert was separated and purified.
Ligate and fragment this,
Blunt the ends of the DNA fragment with T4 polymerase,
A DNA fragment having a length of about 400 bp was recovered. The obtained DNA fragment was used as a plasmid vector, BLUESCRIPT II (S
The product was cloned into the SmaI site (sold by Tratagene) and transformed into E. coli. 20 colonies were picked up randomly and plasmid DN
After preparing A, DNA sequencing was performed on these 20 plasmids (all of which have an ST-NT3 cDNA fragment as an insert). D
The sequencing of NA and the reading of the sequence were performed by the method described in Example 4.

The sequence data of the cDNA fragment of ST-NT3 was edited into a continuous sequence by using the DNA sequence ligation program of DNASIS to obtain the nucleotide sequence shown in SEQ ID NO: 3. An open reading frame was determined from this cDNA full-length sequence data and further translated into an amino acid sequence to obtain the sequence shown in SEQ ID NO: 1.

Example 7: Confirmation of Source of cDNA Encoding Human ST-NT3 Peptide Human glioblastoma cell line T98G (ATCC strain number,
CRL-1690) was homogenized and incubated with oligo dT-cellulose. After cleaning unnecessary materials
Poly A-RNA was eluted and collected (Vennstorm,
B. et. Al., Cell, 28 , 135 (1982)). This RN
A2μg was electrophoresed on 1.0% agarose gel and then blotted on a nitrocellulose membrane to prepare 3 probes.
When 2P-labeled mouse ST-NT3 cDNA was hybridized, the expression of mRNA of about 2.4 kb was observed.

Example 8: Separation and Purification of Human mRNA One 3 × 10 ⁷ human glioblastoma cell line T98G was used.
After stimulating with 100 units / ml of human IL-1β for 4 hours, the method of Okayama et al. (Method in Enzymology, 154 , 3
(1987)). That is, after solubilizing the cells with a 5.5 M GTC solution (5.5 metaganidin thiocyanate, 25 mM sodium citrate, 0.5% sodium lauryl sarcosine), cell lysate (cell lysate) was added at a density of 1.51 to cesium trifluoroacetate (cell lysate). Place on a cushion of CsTFA) solution and perform ultracentrifugation (120,000 xg,
20 hours) and 1.26 mg of total RNA was recovered during the precipitation. This was passed twice through an oligo dT cellulose column to recover 46 μg of Poly (A) ⁺ RNA.

Example 9: Preparation of human cDNA library The cDNA library was prepared by the Gubler & Hoffmann method (Gen
e, 25, 263 (1983)). From Poly (A) ⁺ RNA (5 μg) prepared in Example 8, No
First strand was synthesized by reverse transcriptase using an oligo dT primer having a tI site. Subsequently, second strands were synthesized, ligated with SalI adapter and digested with NotI, followed by gel filtration column chromatography (Sephacryl S-500HR).
(Sold by Pharmacia) were used to remove the adapter and the primer, and 820 ng of the cDNA fraction was recovered. The above cDNA synthesis steps were performed using a kit of SuperScript Lambda System (SuperScript System, sold by BRL).

Example 10: Human ST by cross-hybridization from a human cDNA library
-Screening of NT3 cDNA Recombinant phages were prepared as described in Example 5 to obtain a cDNA library consisting of 1 million plaques. Blotting 1 million plaques obtained on LB plates onto a nitrocellulose membrane,
Eighty positive clones were obtained using ³² P-labeled mouse ST-NT3 cDNA (the same fragment as that prepared in Example 5 and shown in SEQ ID NO: 3) as a probe.

Example 11: Human ST-NT3 cDNA
Isolation of positive clones of
After digestion with NotI and SalI, the length of the insert cDNA was confirmed by agarose gel electrophoresis. 2 out of 5 clones were 1.9 kb, 2 clones were 2.1 k
b, 1 clone was 2.7 kb. From the results of Northern analysis, this cDNA was considered to be almost the entire length of human ST-NT3. Therefore, these three lengths of cDNA digested with NotI and SalI were subjected to agarose gel electrophoresis and then cut out to obtain NotI.pI of pUCCSRaML2.
Subcloned into SalI site
Named β and γ.

Example 12: Human ST-NT3 cDNA
From these full length cDNA sequencing data, the open reading frame (SEQ ID NO: 6, 10 and 1
4) and translated into an amino acid sequence,
The sequences shown in SEQ ID NOs: 5, 9 and 13 were obtained. For the N-terminal 30 to 40 amino acids of the obtained amino acid sequence,
By comparing with the known signal peptide, the signal peptide part in this peptide was estimated (Von Heua
ne, G. Nucleic Acids Res. 14 , 4683 (1986)),
The sequences shown in SEQ ID NOs: 8, 12 and 16 were obtained.

The nucleotide sequence of the DNA was determined by the cycle sequence method using a fluorescent dye terminator (sold by Applied Biosystems Inc.). For sequence reading, a DNA sequencer (Model 373A, Applie
d Biosystems Inc.).

[0099]

[Sequence list]

 SEQ ID NO: 1 Sequence length: 361 Sequence type: Amino acid Topology: Linear Sequence type: Protein sequence Met Val Trp Leu Val Ala Met Thr Pro Arg Gln Ser Ser Leu Cys Gly -35 -30 -25 -20 Leu Ala Ala His Gly Leu Trp Phe Leu Gly Leu Val Leu Leu Met Asp -15 -10 -5 Ala Thr Ala Arg Pro Ala Asn His Ser Ser Thr Arg Glu Arg Ala Ala 1 5 10 Asn Arg Glu Glu Asn Glu Ile Met Pro Pro Asp His Leu Asn Gly Val 15 20 25 Lys Leu Glu Met Asp Gly His Leu Asn Lys Asp Phe His Gln Glu Val 30 35 40 45 Phe Leu Gly Lys Asp Met Asp Gly Phe Asp Glu Asp Ser Glu Pro Arg 50 55 60 Arg Ser Arg Arg Lys Leu Met Val Ile Phe Ser Lys Val Asp Val Asn 65 70 75 Thr Asp Arg Arg Ile Ser Ala Lys Glu Met Gln His Trp Ile Met Glu 80 85 90 Lys Thr Ala Glu His Phe Gln Glu Ala Val Lys Glu Asn Lys Leu His 95 100 105 Phe Arg Ala Val Asp Pro Asp Gly Asp Gly His Val Ser Trp Asp Glu 110 115 120 125 Tyr Lys Val Lys Phe Leu Ala Ser Lys Gly His Asn Glu Arg Glu Ile 130 135 140 Ala Glu Ala Ile Lys Asn His Glu Glu Leu Lys Val Asp Glu Glu Thr 145 150 155 Gln Glu Val Leu Gly Asn Leu Arg Asp Arg Trp Tyr Gln Ala Asp Asn 160 165 170 Pro Pro Ala Asp Leu Leu Leu Thr Glu Asp Glu Phe Leu Ser Phe Leu 175 180 185 His Pro Glu His Ser Arg Gly Met Leu Lys Phe Met Val Lys Glu Ile 190 195 200 205 Phe Arg Asp Leu Asp Gln Asp Gly Asp Lys Gln Leu Ser Leu Pro Glu 210 215 220 Phe Ile Ser Leu Pro Val Gly Thr Val Glu Asn Gln Gln Gly Gln Asp 225 230 235 Ile Asp Asp Asn Trp Val Lys Asp Arg Lys Lys Glu Phe Glu Glu Leu 240 245 250 Ile Asp Ser Asn His Asp Gly Ile Val Thr Met Glu Glu Leu Glu Asn 255 260 265 Tyr Met Asp Pro Met Asn Glu Tyr Asn Ala Leu Asn Glu Ala Lys Gln 270 275 280 285 Met Ile Ala Ile Ala Asp Glu Asn Gln Asn His His Leu Glu Pro Glu 290 295 300 Glu Ile Leu Lys Tyr Ser Glu Phe Phe Thr Gly Ser Lys Leu Met Asp 305 310 315 Tyr Ala Arg Asn Val His Glu Glu Phe 320 325

SEQ ID NO: 2 Sequence Length: 1086 Sequence Type: Nucleic Acid Number of Strands: Single Strand Topology: Linear Sequence Type: cDNA to mRNA Sequence ATGGTCTGGC TGGTGGCAAT GACGCCCAGG CAGAGTTCCC TCTGTGGTCT AGCTGCTCAT 60 GGCCTCTGGT TCTTGGGCCT TGGCCTTCTG ATGG TGCCAACCAC 120 TCATCTACTC GGGAAAGAGC GGCTAACAGG GAGGAAAATG AGATCATGCC CCCAGACCAC 180 CTGAATGGGG TGAAGCTGGA GATGGATGGA CACCTCAACA AGGACTTCCA TCAGGAGGTT 240 TTCCTGGGAA AGGACATGGA TGGGTTTGAT GAAGACTCAG AGCCACGGAG AAGCCGGAGG 300 AAGCTGATGG TCATCTTTTC CAAGGTAGAC GTGAACACTG ACCGGAGGAT CAGCGCTAAG 360 GAGATGCAGC ACTGGATTAT GGAGAAAACA GCAGAGCACT TCCAGGAGGC CGTCAAGGAA 420 AACAAACTGC ACTTCAGGGC TGTGGACCCT GACGGTGACG GCCATGTTTC CTGGGATGAA 480 TATAAAGTGA AGTTTTTGGC AAGCAAAGGC CACAATGAAA GGGAGATTGC TGAAGCCATC 540 AAGAACCATG AGGAGCTCAA AGTGGATGAG GAGACACAGG AAGTCCTTGG GAACCTCAGA 600 GACCGATGGT ATCAGGCAGA CAATCCTCCT GCAGACCTGC TGCTGACTGA GGACGAGTTC 660 CTTTCATTCC TTCACCCTGA GCACAGCCGGGGGGCATGCTCA AGTTCATGG T CAAGGAGATC 720 TTTCGGGACT TGGATCAGGA TGGTGATAAG CAGTTATCTC TGCCCGAGTT CATCTCTCTG 780 CCTGTGGGTA CTGTTGAGAA CCAACAAGGC CAAGACATTG ATGACAACTG GGTGAAAGAC 840 AGGAAGAAGG AGTTTGAGGA ACTGATTGAC TCTAACCATG ATGGGATTGT GACCATGGAG 900 GAGCTAGAGA ACTACATGGA CCCCATGAAT GAATACAATG CCCTCAACGA AGCCAAACAG 960 ATGATTGCCA TTGCTGATGA GAACCAGAAC CACCACCTGG AGCCCGAGGA GATCCTCAAG 1020 TACAGTGAGT TCTTCACCGG CAGCAAGCTC ATGGACTATG CCCGCAACGT GCATGAAGAG 1080 TTCTGA 1086

SEQ ID NO: 3 Sequence Length: 1792 Sequence Type: Nucleic Acid Number of Strands: Single Strand Topology: Linear Sequence Type: cDNA to mRNA Sequence CTGCGCGACT CGGACGCTGG AAGGCGCCTG CGCGACTCGA CTCGGACGCT GGAAGGCGCC 60 TGCGCGACTC GGAGACTGTGCGCGGCCGGCT AGCCCCGAGG 120 CGCACCGTGG CTGGGGGTTT CGACTCCCCG GAGCTGATAT CCAGGGCGGC TTTGAAGGAT 180 AGGGCTGGGC CCAGAATGAT TGACACGTGA CCCCGCGCGC CATTCCTATT GGTTGCTTCG 240 CAGAGGCTGC TTCCGCCCAT AAGAAAGATG GCGGCTTCGA GATTCGTACC GGAGGCCGGG 300 TGTTTTTTTC CCCACCGTTC CCGGTTGTCT TATTCGCCCC TTCCCGGAAG AAAGATGGCG 360 GCGGCCTGAG CCGAGTTCTG CGTAAGACTT GGCGGGCGCC CAGACCCGGC GCATCAGAAG 420 CCGGGGAGTC CCATGGTCTG GCTGGTGGCA ATGACGCCCA GGCAGAGTTC CCTCTGTGGT 480 CTAGCTGCTC ATGGCCTCTG GTTCTTGGGC CTTGTCCTTC TGATGGATGC AACCGCTAGA 540 CCTGCCAACC ACTCATCTAC TCGGGAAAGA GCGGCTAACA GGGAGGAAAA TGAGATCATG 600 CCCCCAGACC ACCTGAATGG GGTGAAGCTG GAGATGGATG GACACCTCAA CAAGGACTTC 660 CATCAGGAGG TTTTCCTGGG AAAGGACATG GATGGGTTTG ATGAAGACT C AGAGCCACGG 720 AGAAGCCGGA GGAAGCTGAT GGTCATCTTT TCCAAGGTAG ACGTGAACAC TGACCGGAGG 780 ATCAGCGCTA AGGAGATGCA GCACTGGATT ATGGAGAAAA CAGCAGAGCA CTTCCAGGAG 840 GCCGTCAAGG AAAACAAACT GCACTTCAGG GCTGTGGACC CTGACGGTGA CGGCCATGTT 900 TCCTGGGATG AATATAAAGT GAAGTTTTTG GCAAGCAAAG GCCACAATGA AAGGGAGATT 960 GCTGAAGCCA TCAAGAACCA TGAGGAGCTC AAAGTGGATG AGGAGACACA GGAAGTCCTT 1020 GGGAACCTCA GAGACCGATG GTATCAGGCA GACAATCCTC CTGCAGACCT GCTGCTGACT 1080 GAGGACGAGT TCCTTTCATT CCTTCACCCT GAGCACAGCC GGGGCATGCT CAAGTTCATG 1140 GTCAAGGAGA TCTTTCGGGA CTTGGATCAG GATGGTGATA AGCAGTTATC TCTGCCCGAG 1200 TTCATCTCTC TGCCTGTGGG TACTGTTGAG AACCAACAAG GCCAAGACAT TGATGACAAC 1260 TGGGTGAAAG ACAGGAAGAA GGAGTTTGAG GAACTGATTG ACTCTAACCA TGATGGGATT 1320 GTGACCATGG AGGAGCTAGA GAACTACATG GACCCCATGA ATGAATACAA TGCCCTCAAC 1380 GAAGCCAAAC AGATGATTGC CATTGCTGAT GAGAACCAGA ACCACCACCT GGAGCCCGAG 1440 GAGATCCTCA AGTACAGTGA GTTCTTCACC GGCAGCAAGC TCATGGACTA TGCCCGCAAC 1500 GTGCATGAAG AGTTCTGAGC ATGCCCCTCA GCTGGGGCCA GGTCGCCAGT GTCCTGTCT G 1560 ACCCTTCCCA TGCTTCGAGG AGGCTATGGT CCAGTGAGAC CTGTGGATGG CTGGGTCCAG 1620 ATTGGGTCTA AAATGCTATC TACCCTGAGT CCGGAGGCTT CCATAGGCCC CAAGAAAGTG 1680 CTGTTCCAAA ACCTCCTCCTTCTGCCATCTCTTGTTCATG

SEQ ID NO: 4 Sequence length: 1792 Sequence type: Nucleic acid Number of strands: Single strand Topology: Linear Sequence type: cDNA to mRNA Origin: Organism name: Mouse Cell line: ST2 Sequence features Characteristic symbol: CDS Location: 433..1518 Characteristic determining method: P Characteristic symbol: sig_peptide Existing location: 433..537 Characteristic determining method: S Characteristic symbol: mat_peptide Existing location: 538 ..1518 method to determine the characteristics: S sequence CTGCGCGACT CGGACGCTGG AAGGCGCCTG CGCGACTCGA CTCGGACGCT GGAAGGCGCC 60 TGCGCGACTC GGAGACTGGC GCGGCCGGTT AGGCCTGGTA ACGTGGGAGG AGCCCCGAGG 120 CGCACCGTGG CTGGGGGTTT CGACTCCCCG GAGCTGATAT CCAGGGCGGC TTTGAAGGAT 180 AGGGCTGGGC CCAGAATGAT TGACACGTGA CCCCGCGCGC CATTCCTATT GGTTGCTTCG 240 CAGAGGCTGC TTCCGCCCAT AAGAAAGATG GCGGCTTCGA GATTCGTACC GGAGGCCGGG 300 TGTTTTTTTC CCCACCGTTC CCGGTTGTCT TATTCGCCCC TTCCCGGAAG AAAGATGGCG 360 GCGGCCTGAG CCGAGTTCTG CGTAAGACTT GGC GGGCGCC CAGACCCGGC GCATCAGAAG 420 CCGGGGAGTC CC ATG GTC TGG CTG GTG GCA ATG ACG CCC AGG CAG AGT 468 Met Val Trp Leu Val Ala Met Thr Pro Arg Gln Ser -35 -30 -25 TCC CTC TGT GGT CTA GCT GCT CAT GGC CTC TGG TTC TTG GGC CTT GTC 516 Ser Leu Cys Gly Leu Ala Ala His Gly Leu Trp Phe Leu Gly Leu Val -20 -15 -10 CTT CTG ATG GAT GCA ACC GCT AGA CCT GCC AAC CAC TCA TCT ACT CGG 564 Leu Leu Met Asp Ala Thr Ala Arg Pro Ala Asn His Ser Ser Thr Arg -5 15 GAA AGA GCG GCT AAC AGG GAG GAA AAT GAG ATC ATG CCC CCA GAC CAC 612 Glu Arg Ala Ala Asn Arg Glu Glu Asn Glu Ile Met Pro Pro Asp His 10 15 20 25 CTG AAT GGG GTG AAG CTG GAG ATG GAT GGA CAC CTC AAC AAG GAC TTC 660 Leu Asn Gly Val Lys Leu Glu Met Asp Gly His Leu Asn Lys Asp Phe 30 35 40 CAT CAG GAG GTT TTC CTG GGA AAG GAC ATG GAT GGG TTT GAT GAA GAC 708 His Gln Glu Val Phe Leu Gly Lys Asp Met Asp Gly Phe Asp Glu Asp 45 50 55 TCA GAG CCA CGG AGA AGC CGG AGG AAG CTG ATG GTC ATC TTT TCC AAG 756 Ser Glu Pro Arg Arg Ser Arg Arg Lys Leu Met Val Ile Phe Ser Lys 60 65 70 GTA GAC GTG AAC ACT GAC CGG AGG ATC AGC GCT AAG GAG ATG CAG CAC 804 Val Asp Val Asn Thr Asp Arg Arg Ile Ser Ala Lys Glu Met Gln His 75 80 85 TGG ATT ATG GAG AAA ACA GCA GAG CAC TTC CAG GAG GCC GTC AAG GAA 852 Trp Ile Met Glu Lys Thr Ala Glu His Phe Gln Glu Ala Val Lys Glu 90 95 100 105 AAC AAA CTG CAC TTC AGG GCT GTG GAC CCT GAC GGT GAC GGC CAT GTT 900 Asn Lys Leu His Phe Arg Ala Val Asp Pro Asp Gly Asp Gly His Val 110 115 120 TCC TGG GAT GAA TAT AAA GTG AAG TTT TTG GCA AGC AAA GGC CAC AAT 948 Ser Trp Asp Glu Tyr Lys Val Lys Phe Leu Ala Ser Lys Gly His Asn 125 130 135 GAA AGG GAG ATT GCT GAA GCC ATC AAG AAC CAT GAG GAG CTC AAA GTG 996 Glu Arg Glu Ile Ala Glu Ala Ile Lys Asn His Glu Glu Leu Lys Val 140 145 150 GAT GAG GAG ACA CAG GAA GTC CTT GGG AAC CTC AGA GAC CGA TGG TAT 1044 Asp Glu Glu Thr Gln Glu Val Leu Gly Asn Leu Arg Asp Arg Trp Tyr 155 160 165 CAG GCA GAC AAT CCT CCT GCA GAC CTG CTG CTG ACT GAG GAC GAG TTC 1092 Gln Ala Asp Asn Pro Pro Ala Asp Leu Leu Leu Thr Glu Asp Glu Phe 170 175 180 185 CTT TCA TTC CTT CAC CCT GAG CAC AGC CGG GGC ATG CTC AAG TTC ATG 1140 Leu Ser Phe Leu His Pro Glu His Ser Arg Gly Met Leu Lys Phe Met 190 195 200 GTC AAG GAG ATC TTT CGG GAC TTG GAT CAG GAT GGT GAT AAG CAG TTA 1188 Val Lys Glu Ile Phe Arg Asp Leu Asp Gln Asp Gly Asp Lys Gln Leu 205 210 215 TCT CTG CCC GAG TTC ATC TCT CTG CCT GTG GGT ACT GTT GAG AAC CAA 1236 Ser Leu Pro Glu Phe Ile Ser Leu Pro Val Gly Thr Val Glu Asn Gln 220 225 230 CAA GGC CAA GAC ATT GAT GAC AAC TGG GTG AAA GAC AGG AAG AAG GAG 1284 Gln Gly Gln Asp Ile Asp Asp Asn Trp Val Lys Asp Arg Lys Lys Glu 235 240 245 TTT GAG GAA CTG ATT GAC TCT AAC CAT GAT GGG ATT GTG ACC ATG GAG 1332 Phe Glu Glu Leu Ile Asp Ser Asn His Asp Gly Ile Val Thr Met Glu 250 255 260 265 GAG CTA GAG AAC TAC ATG GAC CCC ATG AAT GAA TAC AAT GCC CTC AAC 1380 Glu Leu Glu Asn Tyr Met Asp Pro Met Asn Glu Tyr Asn Ala Leu Asn 270 275 280 GAA GCC AAA CAG ATG ATT GCC ATT GCT GAT GAG AAC CAG AAC CAC CAC 1428 Glu Ala Lys Gln Met Ile Ala Ile Ala Ile Ala la Asp Glu Asn Gln Asn His His 285 290 295 CTG GAG CCC GAG GAG ATC CTC AAG TAC AGT GAG TTC TTC ACC GGC AGC 1476 Leu Glu Pro Glu Glu Ile Leu Lys Tyr Ser Glu Phe Phe Thr Gly Ser 300 305 310 AAG CTC ATG GAC TAT GCC CGC AAC GTG CAT GAA GAG TTC TGA 1518 Lys Leu Met Asp Tyr Ala Arg Asn Val His Glu Glu Phe * 315 320 325 GCATGCCCCT CAGCTGGGGC CAGGTCGCCA GTGTCCTGTC TGACCCTTCC CATGCTTCGA 1578 GGAGGCTATG GTCCAGTGAG ACCTGTGGAT GGCTGGGTCC AGATTGGGTC TAAAATGCTA 1638 TCTACCCTGA GTCCGGAGGC TTCCATAGGC CCCAAGAAAG TGCTGTTCCA GAACCTCCTC 1698 CTTCTGTTGT GTCCATATGC TCTGGCTTAG CTCCTGCCAT GGCATGCCCC AGGTTTATTA 1758 AAGATATGCT TAATAGGCAA AAAAAAAAAA AAAA 1792

SEQ ID NO: 5 Sequence length: 362 Sequence type: Amino acid Topology: Linear Sequence type: Protein sequence Met Val Trp Pro Trp Val Ala Met Ala Ser Arg Trp Gly Pro Leu Ile -36 -35- 30 -25 Gly Leu Ala Pro Cys Cys Leu Trp Leu Leu Gly Ala Val Leu Leu Met -20 -15 -10 -5 Asp Ala Ser Ala Arg Pro Ala Asn His Ser Ser Thr Arg Glu Arg Val 1 5 10 Ala Asn Arg Glu Glu Asn Glu Ile Leu Pro Pro Asp His Leu Asn Gly 15 20 25 Val Lys Leu Glu Met Asp Gly His Leu Asn Arg Gly Phe His Gln Glu 30 35 40 Val Phe Leu Gly Lys Asp Leu Gly Gly Phe Asp Glu Asp Ala Glu Pro 45 50 55 60 Arg Arg Ser Arg Arg Lys Leu Met Val Ile Phe Ser Lys Val Asp Val 65 70 75 Asn Thr Asp Arg Lys Ile Ser Ala Lys Glu Met Gln Arg Trp Ile Met 80 85 90 Glu Lys Thr Ala Glu His Phe Gln Glu Ala Met Glu Glu Ser Lys Thr 95 100 105 His Phe Arg Ala Val Asp Pro Asp Gly Asp Gly His Val Ser Trp Asp 110 115 120 Glu Tyr Lys Val Lys Phe Leu Ala Ser Lys Gly His Ser Glu Lys Glu 125 130 135 140 Val Ala A sp Ala Ile Arg Leu Asn Glu Glu Leu Lys Val Asp Glu Glu 145 150 155 Thr Gln Glu Val Leu Glu Asn Leu Lys Asp Arg Trp Tyr Gln Ala Asp 160 165 170 Ser Pro Pro Ala Asp Leu Leu Leu Thr Glu Glu Glu Phe Leu Ser Phe 175 180 185 Leu His Pro Glu His Ser Arg Gly Met Leu Arg Phe Met Val Lys Glu 190 195 200 Ile Val Arg Asp Leu Asp Gln Asp Gly Asp Lys Gln Leu Ser Val Pro 205 210 215 220 Glu Phe Ile Ser Leu Pro Val Ala Thr Val Glu Asn Gln Gln Gly Gln 225 230 235 Asp Ile Asp Asp Asn Trp Val Lys Asp Arg Lys Lys Glu Phe Glu Glu 240 245 250 Leu Ile Asp Ser Asn His Asp Gly Ile Val Thr Ala Glu Glu Leu Glu 255 260 265 Ser Tyr Met Asp Pro Met Asn Glu Tyr Asn Ala Leu Asn Glu Pro Lys 270 275 275 280 Gln Met Ile Ala Val Ala Asp Glu Asn Gln Asn His His Leu Glu Pro 285 290 295 300 Glu Glu Val Leu Lys Tyr Ser Glu Phe Phe Thr Gly Ser Lys Leu Val 305 310 315 Asp Tyr Ala Arg Ser Val His Glu Glu Phe 320 325

SEQ ID NO: 6 Sequence Length: 1089 Sequence Type: Nucleic Acid Number of Strands: Single Strand Topology: Linear Sequence Type: cDNA to mRNA Sequence ATGGTCTGGC CCTGGGTGGC GATGGCGTCC AGGTGGGGTC CCCTCATTGG CCTGGCTCCG 60 TGCTGCCTCT GGCTCCTGGG GGCACGCCTT CTGA GCCTGCCAAC 120 CACTCGTCCA CTCGAGAGAG AGTAGCCAAC AGGGAGGAGA ATGAGATCCT GCCCCCAGAC 180 CACCTGAACG GGGTGAAGCT GGAGATGGAC GGGCACCTCA ATCGCGGCTT CCACCAGGAG 240 GTCTTCCTAG GCAAGGACCT GGGTGGCTTT GATGAGGACG CGGAGCCGCG GCGGAGCCGG 300 AGGAAGCTGA TGGTCATCTT TTCCAAGGTG GATGTGAACA CTGACCGGAA GATCAGTGCC 360 AAGGAGATGC AGCGCTGGAT CATGGAGAAG ACGGCCGAGC ACTTCCAGGA GGCCATGGAG 420 GAGAGCAAGA CACACTTCCG CGCCGTGGAC CCTGACGGGG ACGGTCACGT GTCTTGGGAC 480 GAGTATAAGG TGAAGTTTTT GGCGAGTAAA GGCCATAGCG AGAAGGAGGT TGCCGACGCC 540 ATCAGGCTCA ACGAGGAACT CAAAGTGGAT GAGGAAACAC AGGAAGTCCT GGAGAACCTG 600 AAGGACCGCT GGTACCAGGC GGACAGCCCC CCTGCAGACC TGCTGCTGAC GGAGGAGGAG 660 TTCCTGTCGT TCCTCCACCC CGAGCACAGC CGGGGAATGC TCAGGTTCA T GGTGAAGGAG 720 ATCGTCCGGG ACCTGGACCA GGACGGTGAC AAGCAGCTCT CTGTGCCCGA GTTCATCTCC 780 CTGCCCGTGG CCACCGTGGA GAACCAGCAG GGCCAGGACA TTGACGACAA CTGGGTGAAA 840 GACAGAAAAA AGGAGTTTGA GGAGCTCATT GACTCCAACC ACGACGGCAT CGTGACCGCC 900 GAGGAGCTGG AGAGCTACAT GGACCCCATG AACGAGTACA ACGCGCTGAA CGAGCCCAAG 960 CAGATGATCG CCGTCGCCGA CGAGAACCAG AACCACCACC TGGAGCCCGA GGAGGTGCTC 1020 AAGTACAGCG AGTTCTTCAC GGGCAGCAAG CTGGTGGACT ACGCGCGCAG CGTGCACGAG 1080 GAGTTTTGA 1089

SEQ ID NO: 7 Sequence Length: 1912 Sequence Type: Nucleic Acid Number of Strands: Single Strand Topology: Linear Sequence Type: cDNA to mRNA Sequence CCTGATCCGG GCCCGTTGGCGCGGCGTCACTG ACGCTTCGCT CCGGTCCTCG GATCCCGAGC 60 GCGGGGAGGC AGACCGACTG TGTCCCCGGTCGCCCATGTCGCCCATCATCCGGTCGCCCATGTCCGACAT CTGGGGACAC 120 AGAGCCCTCC GTGGTGCCCG GGGATTGGAT TGGAGCCAGG ACCTCACTTC CTCCTCTGCC 180 CCTGCCCCTG CCCCTCCCAG CACCTGGCCC ACACCCTGCA GCCCGCCCCA TGGTCTGGCC 240 CTGGGTGGCG ATGGCGTCCA GGTGGGGTCC CCTCATTGGC CTGGCTCCGT GCTGCCTCTG 300 GCTCCTGGGG GCAGTCCTTC TGATGGACGC GTCTGCACGG CCTGCCAACC ACTCGTCCAC 360 TCGAGAGAGA GTAGCCAACA GGGAGGAGAA TGAGATCCTG CCCCCAGACC ACCTGAACGG 420 GGTGAAGCTG GAGATGGACG GGCACCTCAA TCGCGGCTTC CACCAGGAGG TCTTCCTAGG 480 CAAGGACCTG GGTGGCTTTG ATGAGGACGC GGAGCCGCGG CGGAGCCGGA GGAAGCTGAT 540 GGTCATCTTT TCCAAGGTGG ATGTGAACAC TGACCGGAAG ATCAGTGCCA AGGAGATGCA 600 GCGCTGGATC ATGGAGAAGA CGGCCGAGCA CTTCCAGGAG GCCATGGAGG AGAGCAAGAC 660 ACACTTCCGC GCCGTGGACC CTGACGGGGA CGGTCACGTG TCTTGGGAC G AGTATAAGGT 720 GAAGTTTTTG GCGAGTAAAG GCCATAGCGA GAAGGAGGTT GCCGACGCCA TCAGGCTCAA 780 CGAGGAACTC AAAGTGGATG AGGAAACACA GGAAGTCCTG GAGAACCTGA AGGACCGCTG 840 GTACCAGGCG GACAGCCCCC CTGCAGACCT GCTGCTGACG GAGGAGGAGT TCCTGTCGTT 900 CCTCCACCCC GAGCACAGCC GGGGAATGCT CAGGTTCATG GTGAAGGAGA TCGTCCGGGA 960 CCTGGACCAG GACGGTGACA AGCAGCTCTC TGTGCCCGAG TTCATCTCCC TGCCCGTGGC 1020 CACCGTGGAG AACCAGCAGG GCCAGGACAT TGACGACAAC TGGGTGAAAG ACAGAAAAAA 1080 GGAGTTTGAG GAGCTCATTG ACTCCAACCA CGACGGCATC GTGACCGCCG AGGAGCTGGA 1140 GAGCTACATG GACCCCATGA ACGAGTACAA CGCGCTGAAC GAGCCCAAGC AGATGATCGC 1200 CGTCGCCGAC GAGAACCAGA ACCACCACCT GGAGCCCGAG GAGGTGCTCA AGTACAGCGA 1260 GTTCTTCACG GGCAGCAAGC TGGTGGACTA CGCGCGCAGC GTGCACGAGG AGTTTTGAGC 1320 GCCCGGCCGC GCCCCGCGCC GCCCCCCACG CACCACCGGG GGGGCCTCGC GGGTGACTCC 1380 GGCCTCCGTG GCTGTCCCGG ACCCCACCTC TTCTCTGCCG CCCACCACCG GCCGACCGAC 1440 CGCGGCTTCC CCAGTTGATG AGCGGCGTGT CCCCTCTGCA GCGCGCACCC CGGCGGGGCT 1500 TTGGCTGTGA CGCGGTCGGG GCGCGGGGCT GGTCTGTGGC CCCGCGGCGC GCCTCCTCC C 1560 TGGTCCCTCG AAATCGTGGC ATCTCACTTC TGAGAACGAA ATCTCGCTTC AGTCACTCTG 1620 CCGAAGGCGC TGACGGCATC GCGGCCGGAA CCTCTGGGCC CGGCCCCTCC CAGGGCCGCC 1680 GCTCCGTGGG AAAAAACAGC TCCTCCATTT CCTTGGAAAC TGAACGATTA TTAAAAATAG 1740 ATAAACTTCG CTGGAAATGA GTAGCCAGGA AGTTCAGGGG AGGGTCCGGG GTCCTTCCCG 1800 GGGCCTGGCG TGTCGGAACC ACCCAGGTCC CGCAGCTGCC TCTGAGAAAA TCCAAATATT 1860 TTTTGTGACA AGAATCACAA ACATTTACTT TAAATATAAA AAAAAAAAAA AA 1912

SEQ ID NO: 8 Sequence length: 1912 Sequence type: Nucleic acid Number of strands: Single strand Topology: Linear Sequence type: cDNA to mRNA Origin: Organism name: Homo Sapiens Cell line: T98G Sequence Feature Symbol representing the feature: CDS Location: 230..1318 Method by which the feature was determined: P Symbol representing the feature: sig_peptide Location: 230..337 Method by which the feature was determined: S Symbol representing the feature: mat_peptide Location: 338..1318 method to determine the characteristics: S sequence CCTGATCCGG GCCCGTTGGC GGCGTCACTG ACGCTTCGCT CCGGTCCTCG GATCCCGAGC 60 GCGGGGAGGC AGACCGACTG TGAGCTGCTT GTCCCCATCC TGCGGACGTC CTGGGGACAC 120 AGAGCCCTCC GTGGTGCCCG GGGATTGGAT TGGAGCCAGG ACCTCACTTC CTCCTCTGCC 180 CCTGCCCCTG CCCCTCCCAG CACCTGGCCC ACACCCTGCA GCCCGCCCC ATG GTC 235 Met Val -36 -35 TGG CCC TGG GTG GCG ATG GCG TCC AGG TGG GGT CCC CTC ATT GGC CTG 283 Trp Pro Trp Val Ala Met Ala Ser Arg Trp Gly Pro Leu Ile Gly Leu -30 -25 -20 GCT CCG TGC TG C CTC TGG CTC CTG GGG GCA GTC CTT CTG ATG GAC GCG 331 Ala Pro Cys Cys Leu Trp Leu Leu Gly Ala Val Leu Leu Met Asp Ala -15 -10 -5 TCT GCA CGG CCT GCC AAC CAC TCG TCC ACT CGA GAG AGA GTA GCC AAC 379 Ser Ala Arg Pro Ala Asn His Ser Ser Thr Arg Glu Arg Val Ala Asn 1 5 10 AGG GAG GAG AAT GAG ATC CTG CCC CCA GAC CAC CTG AAC GGG GTG AAG 427 Arg Glu Glu Asn Glu Ile Leu Pro Pro Asp His Leu Asn Gly Val Lys 15 20 25 30 CTG GAG ATG GAC GGG CAC CTC AAT CGC GGC TTC CAC CAG GAG GTC TTC 475 Leu Glu Met Asp Gly His Leu Asn Arg Gly Phe His Gln Glu Val Phe 35 40 45 CTA GGC AAG GAC CTG GGT GGC TTT GAT GAG GAC GCG GAG CCG CGG CGG 523 Leu Gly Lys Asp Leu Gly Gly Phe Asp Glu Asp Ala Glu Pro Arg Arg 50 55 60 AGC CGG AGG AAG CTG ATG GTC ATC TTT TCC AAG GTG GAT GTG AAC ACT 571 Ser Arg Arg Lys Leu Met Val Ile Phe Ser Lys Val Asp Val Asn Thr 65 70 75 GAC CGG AAG ATC AGT GCC AAG GAG ATG CAG CGC TGG ATC ATG GAG AAG 619 Asp Arg Lys Ile Ser Ala Lys Glu Met Gln Arg Trp Ile Met Glu Lys 80 85 90 ACG GCC GAG CAC TTC CAG GAG GCC ATG GAG GAG AGC AAG ACA CAC TTC 667 Thr Ala Glu His Phe Gln Glu Ala Met Glu Glu Ser Lys Thr His Phe 95 100 105 110 CGC GCC GTG GAC CCT GAC GGG GAC GGT CAC GTG TCT TGG GAC GAG TAT 715 Arg Ala Val Asp Pro Asp Gly Asp Gly His Val Ser Trp Asp Glu Tyr 115 120 125 AAG GTG AAG TTT TTG GCG AGT AAA GGC CAT AGC GAG AAG GAG GTT GCC 763 Lys Val Lys Phe Leu Ala Ser Lys Gly His Ser Glu Lys Glu Val Ala 130 135 140 GAC GCC ATC AGG CTC AAC GAG GAA CTC AAA GTG GAT GAG GAA ACA CAG 811 Asp Ala Ile Arg Leu Asn Glu Glu Leu Lys Val Asp Glu Glu Thr Gln 145 150 155 GAA GTC CTG GAG AAC CTG AAG GAC CGC TGG TAC CAG GCG GAC AGC CCC 859 Glu Val Leu Glu Asn Leu Lys Asp Arg Trp Tyr Gln Ala Asp Ser Pro 160 165 170 CCT GCA GAC CTG CTG CTG ACG GAG GAG GAG TTC CTG TCG TTC CTC CAC 907 Pro Ala Asp Leu Leu Leu Thr Glu Glu Glu Phe Leu Ser Phe Leu His 175 180 185 190 CCC GAG CAC AGC CGG GGA ATG CTC AGG TTC ATG GTG AAG GAG ATC GTC 955 Pro Glu His Ser Arg Gly Met Leu Arg Phe Met Val Lys Glu Ile Val 195 200 205 CGG GACCTG GAC CAG GAC GGT GAC AAG CAG CTC TCT GTG CCC GAG TTC 1003 Arg Asp Leu Asp Gln Asp Gly Asp Lys Gln Leu Ser Val Pro Glu Phe 210 215 220 ATC TCC CTG CCC GTG GCC ACC GTG GAG AAC CAG CAG GGC CAG GAC ATT 1051 Ile Ser Leu Pro Val Ala Thr Val Glu Asn Gln Gln Gly Gln Asp Ile 225 230 235 GAC GAC AAC TGG GTG AAA GAC AGA AAA AAG GAG TTT GAG GAG CTC ATT 1099 Asp Asp Asn Trp Val Lys Asp Arg Lys Lys Glu Phe Glu Glu Leu Ile 240 245 250 GAC TCC AAC CAC GAC GGC ATC GTG ACC GCC GAG GAG CTG GAG AGC TAC 1147 Asp Ser Asn His Asp Gly Ile Val Thr Ala Glu Glu Leu Glu Ser Tyr 255 260 265 270 ATG GAC CCC ATG AAC GAG TAC AAC GCG CTG AAC GAG CCC AAG CAG ATG 1195 Met Asp Pro Met Asn Glu Tyr Asn Ala Leu Asn Glu Pro Lys Gln Met 275 280 285 ATC GCC GTC GCC GAC GAG AAC CAG AAC CAC CAC CTG GAG CCC GAG GAG 1243 Ile Ala Val Ala Asp Glu Asn Gln Asn His His Leu Glu Pro Glu Glu 290 295 300 GTG CTC AAG TAC AGC GAG TTC TTC ACG GGC AGC AAG CTG GTG GAC TAC 1291 Val Leu Lys Tyr Ser Glu Phe Phe Thr Gly Ser Lys Leu Val Asp Tyr 3 05 310 315 GCG CGC AGC GTG CAC GAG GAG TTT TGA GCGCCCGGCC GCGCCCCGCG 1338 Ala Arg Ser Val His Glu Glu Phe * 320 325 CCGCCCCCCA CGCACCACCG GGGGGGCCTC GCGGGTGACT CCGGCCTCCG TGGCTGTCCC 1398 GGACCCCACC TCTTCTCTGC CGCCCACCAC CGGCCGACCG ACCGCGGCTT CCCCAGTTGA 1458 TGAGCGGCGT GTCCCCTCTG CAGCGCGCAC CCCGGCGGGG CTTTGGCTGT GACGCGGTCG 1518 GGGCGCGGGG CTGGTCTGTG GCCCCGCGGC GCGCCTCCTC CCTGGTCCCT CGAAATCGTG 1578 GCATCTCACT TCTGAGAACG AAATCTCGCT TCAGTCACTC TGCCGAAGGC GCTGACGGCA 1638 TCGCGGCCGG AACCTCTGGG CCCGGCCCCT CCCAGGGCCG CCGCTCCGTG GGAAAAAACA 1698 GCTCCTCCAT TTCCTTGGAA ACTGAACGAT TATTAAAAAT AGATAAACTT CGCTGGAAAT 1758 GAGTAGCCAG GAAGTTCAGG GGAGGGTCCG GGGTCCTTCC CGGGGCCTGG CGTGTCGGAA 1818 CCACCCAGGT CCCGCAGCTG CCTCTGAGAA AATCCAAATA TTTTTTGTGA CAAGAATCAC 1878 AAACATTTAC TTTAAATATA AAAAAAAAAA AAAA 1912

SEQ ID NO: 9 Sequence length: 254 Sequence type: Amino acid Topology: Linear Sequence type: Protein sequence Met Val Trp Pro Trp Val Ala Met Ala Ser Arg Trp Gly Pro Leu Ile -36 -35- 30 -25 Gly Leu Ala Pro Cys Cys Leu Trp Leu Leu Gly Ala Val Leu Leu Met -20 -15 -10 -5 Asp Ala Ser Ala Arg Pro Ala Asn His Ser Ser Thr Arg Glu Arg Val 1 5 10 Ala Asn Arg Glu Glu Asn Glu Ile Leu Pro Pro Asp His Leu Asn Gly 15 20 25 Val Lys Leu Glu Met Asp Gly His Leu Asn Arg Gly Phe His Gln Glu 30 35 40 Val Phe Leu Gly Lys Asp Leu Gly Gly Phe Asp Glu Asp Ala Glu Pro 45 50 55 60 Arg Arg Ser Arg Arg Lys Leu Met Val Ile Phe Ser Lys Val Asp Val 65 70 75 Asn Thr Asp Arg Lys Ile Ser Ala Lys Glu Met Gln Arg Trp Ile Met 80 85 90 Glu Lys Thr Ala Glu His Phe Gln Glu Ala Met Glu Glu Ser Lys Thr 95 100 105 His Phe Arg Ala Val Asp Pro Asp Gly Asp Gly His Val Ser Trp Asp 110 115 120 Glu Tyr Lys Val Lys Phe Leu Ala Ser Lys Gly His Ser Glu Lys Glu 125 130 135 140 Val Ala A sp Ala Ile Arg Leu Asn Glu Glu Leu Lys Val Asp Glu Glu 145 150 155 Ser Cys Ala Pro Leu Ser Thr Gly Ser Pro Gly Glu Pro Glu Gly Pro 160 165 170 Leu Val Pro Gly Gly Gln Pro Pro Cys Arg Pro Ala Ala Asp Gly Gly 175 180 185 Gly Val Pro Val Gly Pro Pro Pro Arg Ala Gln Pro Gly Asn Ala Gln 190 195 200 Val His Gly Glu Gly Asp Arg Pro Gly Pro Gly Pro Gly Arg 205 210 215

SEQ ID NO: 10 Sequence length: 765 Sequence type: Nucleic acid Number of strands: Single strand Topology: Linear Sequence type: cDNA to mRNA Sequence ATGGTCTGGC CCTGGGTGGC GATGGCGTCC AGGTGGGGTC CCCTCATTGG CCTGGCTCCG 60 TGCTGCCTCT GGCTCCTGGG GGCACGCCTT CTGA GCCTGCCAAC 120 CACTCGTCCA CTCGAGAGAG AGTAGCCAAC AGGGAGGAGA ATGAGATCCT GCCCCCAGAC 180 CACCTGAACG GGGTGAAGCT GGAGATGGAC GGGCACCTCA ATCGCGGCTT CCACCAGGAG 240 GTCTTCCTAG GCAAGGACCT GGGTGGCTTT GATGAGGACG CGGAGCCGCG GCGGAGCCGG 300 AGGAAGCTGA TGGTCATCTT TTCCAAGGTG GATGTGAACA CTGACCGGAA GATCAGTGCC 360 AAGGAGATGC AGCGCTGGAT CATGGAGAAG ACGGCCGAGC ACTTCCAGGA GGCCATGGAG 420 GAGAGCAAGA CACACTTCCG CGCCGTGGAC CCTGACGGGG ACGGTCACGT GTCTTGGGAC 480 GAGTATAAGG TGAAGTTTTT GGCGAGTAAA GGCCATAGCG AGAAGGAGGT TGCCGACGCC 540 ATCAGGCTCA ACGAGGAACT CAAAGTGGAT GAGGAAAGCT GCGCCCCTCT CAGCACAGGA 600 AGTCCTGGAG AACCTGAAGG ACCGCTGGTA CCAGGCGGAC AGCCCCCCTG CAGACCTGCT 660 GCTGACGGAG GAGGAGTTCC TGTCGGTCCT CCACCCCGAG CACAGCCGG G GAATGCTCAG 720 GTTCATGGTG AAGGAGATCG TCCGGGACCT GGACCAGGAC GGTGA 765

SEQ ID NO: 11 Sequence length: 2114 Sequence type: Nucleic acid Number of strands: Single-strand topology: Linear Sequence type: cDNA to mRNA Sequence GGCGCCGGCG TCCCCTCCGTGAGGTCGCGC CCGTTCGCAC CGCCCCCGCC CGCAAGAAAG 60 ATGGCAGTGG CCTGATCCGGCGCCACTTGG GG CCGGTCCTCG 120 GATCCCGAGC GCGGGGAGGC AGACCGACTG TGAGCTGCTT GTCCCCATCC TGCGGACGTC 180 CTGGGGACAC AGAGCCCTCC GTGGTGCCCG GGGATTGGAT TGGAGCCAGG ACCTCACTTC 240 CTCCTCTGCC CCTGCCCCTG CCCCTCCCAG CACCTGGCCC ACACCCTGCA GCCCGCCCCA 300 TGGTCTGGCC CTGGGTGGCG ATGGCGTCCA GGTGGGGTCC CCTCATTGGC CTGGCTCCGT 360 GCTGCCTCTG GCTCCTGGGG GCAGTCCTTC TGATGGACGC GTCTGCACGG CCTGCCAACC 420 ACTCGTCCAC TCGAGAGAGA GTAGCCAACA GGGAGGAGAA TGAGATCCTG CCCCCAGACC 480 ACCTGAACGG GGTGAAGCTG GAGATGGACG GGCACCTCAA TCGCGGCTTC CACCAGGAGG 540 TCTTCCTAGG CAAGGACCTG GGTGGCTTTG ATGAGGACGC GGAGCCGCGG CGGAGCCGGA 600 GGAAGCTGAT GGTCATCTTT TCCAAGGTGG ATGTGAACAC TGACCGGAAG ATCAGTGCCA 660 AGGAGATGCA GCGCTGGATC ATGGAGAAGA CGGCCGAGCA CTTCCA GGAG GCCATGGAGG 720 AGAGCAAGAC ACACTTCCGC GCCGTGGACC CTGACGGGGA CGGTCACGTG TCTTGGGACG 780 AGTATAAGGT GAAGTTTTTG GCGAGTAAAG GCCATAGCGA GAAGGAGGTT GCCGACGCCA 840 TCAGGCTCAA CGAGGAACTC AAAGTGGATG AGGAAAGCTG CGCCCCTCTC AGCACAGGAA 900 GTCCTGGAGA ACCTGAAGGA CCGCTGGTAC CAGGCGGACA GCCCCCCTGC AGACCTGCTG 960 CTGACGGAGG AGGAGTTCCT GTCGGTCCTC CACCCCGAGC ACAGCCGGGG AATGCTCAGG 1020 TTCATGGTGA AGGAGATCGT CCGGGACCTG GACCAGGACG GTGACAAGCA GCTCTCTGTG 1080 CCCGAGTTCA TCTCCCTGCC CGTGGGCACC GTGGAGAACC AGCAGGGCCA GGACATTGAC 1140 GACAACTGGG TGAAAGACAG AAAAAAGGAG TTTGAGGAGC TCATTGACTC CAACCACGAC 1200 GGCATCGTGA CCGCCGAGGA GCTGGAGAAC GTGCCCACAC TCCCGCTGCA GCCAATAGGC 1260 ACCTTAAATA GCCACTTCGT GCGGCTGGCC GCGGAGCTCG GAGGGGGAAA GGCGACGCTG 1320 ACCTGTGCCC CGCTCGCCCG CAGAGCTACA TGGACCCCAT GAACGAGTAC AACGCGCTGA 1380 ACGAGGCCAA GCAGATGATC GCCGTCGCCG ACGAGAACCA GAACCACCAC CTGGAGCCCG 1440 AGGAGGTGCT CAAGTACAGC GAGTTCTTCA CGGGCAGCAA GCTGGTGGAC TACGCGCGCA 1500 GCGTGCACGA GGAGTTTTGA GCGCCCGGCC GCGCCCCGCG CCGCCCCCCA CGCACC ACCG 1560 GGGGGGCCTC GCGGGTGACT CCGGCCTCCG TGGCTGTCCC GGACCCCACC TCTTCTCTGC 1620 CGCCCACCAC CGGCCGACCG ACCGCGGCTT CCCCAGTTGA TGAGCGGCGT GTCCCCTCTG 1680 CAGCGCGCAC CCCGGCGGGG CTTTGGCTGT GACGCGGTCG GGGCGCGGGG CTGGTCTGTG 1740 GCCCCGCGGC GCGCCTCCTC CCTGGTCCCT CGAAATCGTG GCATCTCACT TCTGAGAACG 1800 AAATCTCGCT TCAGTCACTC TGCCGAAGGC GCTGACGGCA TCGCGGCCGG AACCTCTGGG 1860 CCCGGCCCCT CCCAGGGCCG CCGCTCCGTG GGAAAAAACA GCTCCTCCAT TTCCTTGGAA 1920 ACTGAACGAT TATTAAAAAT AGATAAACTT CGCTGGAAAT GAGTAGCCAG GAAGTTCAGG 1980 GGAGGGTCCG GGGTCCTTCC CGGGGCCTGG CGTGTCGGAA CCACCCAGGT CCCGCAGCTG 2040 CCTCTGAGAA AATCCAAATA TTTTTTGTGA CAAGAATCAC AAACATTTAC TTTAAATATA 2100 AAAAAAAAAA AAAA 2114

SEQ ID NO: 12 Sequence length: 2114 Sequence type: Nucleic acid Number of strands: Single strand Topology: Linear Sequence type: cDNA to mRNA Origin: Organism name: Homo Sapiens Cell line: T98G Sequence Feature Symbol that represents the feature: CDS Location: 300..1064 Method that determines the feature: P Feature symbol: sig_peptide Location: 300..407 Method that determines the feature: S Symbol that represents the feature: mat_peptide Location: 408..1064 method to determine the characteristics: S sequence GGCGCCGGCG TCCCCTCCGT GAGGTCGCGC CCGTTCGCAC CGCCCCCGCC CGCAAGAAAG 60 ATGGCAGTGG CCTGATCCGG GCCCGTTGGC GGCGTCACTG ACGCTTCGCT CCGGTCCTCG 120 GATCCCGAGC GCGGGGAGGC AGACCGACTG TGAGCTGCTT GTCCCCATCC TGCGGACGTC 180 CTGGGGACAC AGAGCCCTCC GTGGTGCCCG GGGATTGGAT TGGAGCCAGG ACCTCACTTC 240 CTCCTCTGCC CCTGCCCCTG CCCCTCCCAG CACCTGGCCC ACACCCTGCA GCCCGCCCC 299 ATG GTC TGG CCC TGG GTG GCG ATG GCG TCC AGG TGG GGT CCC CTC ATT 347 Met Val Trp Pro Trp Val Ala Me t Ala Ser Arg Trp Gly Pro Leu Ile -36 -35 -30 -25 GGC CTG GCT CCG TGC TGC CTC TGG CTC CTG GGG GCA GTC CTT CTG ATG 395 Gly Leu Ala Pro Cys Cys Leu Trp Leu Leu Gly Ala Val Leu Leu Met -20 -15 -10 -5 GAC GCG TCT GCA CGG CCT GCC AAC CAC TCG TCC ACT CGA GAG AGA GTA 443 Asp Ala Ser Ala Arg Pro Ala Asn His Ser Ser Thr Arg Glu Arg Val 1 5 10 GCC AAC AGG GAG GAG AAT GAG ATC CTG CCC CCA GAC CAC CTG AAC GGG 491 Ala Asn Arg Glu Glu Asn Glu Ile Leu Pro Pro Asp His Leu Asn Gly 15 20 25 GTG AAG CTG GAG ATG GAC GGG CAC CTC AAT CGC GGC TTC CAC CAG GAG 539 Val Lys Leu Glu Met Asp Gly His Leu Asn Arg Gly Phe His Gln Glu 30 35 40 GTC TTC CTA GGC AAG GAC CTG GGT GGC TTT GAT GAG GAC GCG GAG CCG 587 Val Phe Leu Gly Lys Asp Leu Gly Gly Phe Asp Glu Asp Ala Glu Pro 45 50 55 60 CGG CGG AGC CGG AGG AAG CTG ATG GTC ATC TTT TCC AAG GTG GAT GTG 635 Arg Arg Ser Arg Arg Lys Leu Met Val Ile Phe Ser Lys Val Asp Val 65 70 75 AAC ACT GAC CGG AAG ATC AGT GCC AAG GAG ATG CAG CGC TGG ATC ATG 683 Asn Thr Asp Arg Lys Ile Ser A la Lys Glu Met Gln Arg Trp Ile Met 80 85 90 GAG AAG ACG GCC GAG CAC TTC CAG GAG GCC ATG GAG GAG AGC AAG ACA 731 Glu Lys Thr Ala Glu His Phe Gln Glu Ala Met Glu Glu Ser Lys Thr 95 100 105 CAC TTC CGC GCC GTG GAC CCT GAC GGG GAC GGT CAC GTG TCT TGG GAC 779 His Phe Arg Ala Val Asp Pro Asp Gly Asp Gly His Val Ser Trp Asp 110 115 120 GAG TAT AAG GTG AAG TTT TTG GCG AGT AAA GGC CAT AGC GAG AAG GAG 827 Glu Tyr Lys Val Lys Phe Leu Ala Ser Lys Gly His Ser Glu Lys Glu 125 130 135 140 GTT GCC GAC GCC ATC AGG CTC AAC GAG GAA CTC AAA GTG GAT GAG GAA 875 Val Ala Asp Ala Ile Arg Leu Asn Glu Glu Leu Lys Val Asp Glu Glu 145 150 155 AGC TGC GCC CCT CTC AGC ACA GGA AGT CCT GGA GAA CCT GAA GGA CCG 923 Ser Cys Ala Pro Leu Ser Thr Gly Ser Pro Gly Glu Pro Glu Gly Pro 160 165 170 CTG GTA CCA GGC GGA CAG CCC CCC TGC AGA CCT GCT GCT GAC GGA GGA 971 Leu Val Pro Gly Gly Gln Pro Pro Cys Arg Pro Ala Ala Asp Gly Gly 175 180 185 GGA GTT CCT GTC GGT CCT CCA CCC CGA GCA CAG CCG GGG AAT GCT CAG 1019 Gly Val Pro Val Gly Pro Pro Pro Arg Ala Gln Pro Gly Asn Ala Gln 190 195 200 GTT CAT GGT GAA GGA GAT CGT CCG GGA CCT GGA CCA GGA CGG TGA 1064 Val His Gly Glu Gly Asp Arg Pro Gly Pro Gly Pro Gly Arg * 205 210 215 CAAGCAGCTC TCTGTGCCCG AGTTCATCTC CCTGCCCGTG GGCACCGTGG AGAACCAGCA 1124 GGGCCAGGAC ATTGACGACA ACTGGGTGAA AGACAGAAAA AAGGAGTTTG AGGAGCTCAT 1184 TGACTCCAAC CACGACGGCA TCGTGACCGC CGAGGAGCTG GAGAACGTGC CCACACTCCC 1244 GCTGCAGCCA ATAGGCACCT TAAATAGCCA CTTCGTGCGG CTGGCCGCGG AGCTCGGAGG 1304 GGGAAAGGCG ACGCTGACCT GTGCCCCGCT CGCCCGCAGA GCTACATGGA CCCCATGAAC 1364 GAGTACAACG CGCTGAACGA GGCCAAGCAG ATGATCGCCG TCGCCGACGA GAACCAGAAC 1424 CACCACCTGG AGCCCGAGGA GGTGCTCAAG TACAGCGAGT TCTTCACGGG CAGCAAGCTG 1484 GTGGACTACG CGCGCAGCGT GCACGAGGAG TTTTGAGCGC CCGGCCGCGC CCCGCGCCGC 1544 CCCCCACGCA CCACCGGGGG GGCCTCGCGG GTGACTCCGG CCTCCGTGGC TGTCCCGGAC 1604 CCCACCTCTTTT CTCTGCCGCC CACCACCGGC CGACCGAGGGG CGGCTTCCCC AGTTGATGAG 1664 CGGCGGGGGCGCCCCCCCAGTTGATGAG 1664 CGGCGGGGGCGCCCCCTCTGCAGCTT CGCGC CTCCTCCCTG GTCCCTCGAA ATCGTGGCAT 1784 CTCACTTCTG AGAACGAAAT CTCGCTTCAG TCACTCTGCC GAAGGCGCTG ACGGCATCGC 1844 GGCCGGAACC TCTGGGCCCG GCCCCTCCCA GGGCCGCCGC TCCGTGGGAA AAAACAGCTC 1904 CTCCATTTCC TTGGAAACTG AACGATTATT AAAAATAGAT AAACTTCGCT GGAAATGAGT 1964 AGCCAGGAAG TTCAGGGGAG GGTCCGGGGT CCTTCCCGGG GCCTGGCGTG TCGGAACCAC 2024 CCAGGTCCCG CAGCTGCCTC TGAGAAAATC CAAATATTTT TTGTGACAAG AATCACAAAC 2084 ATTTACTTTA AATATAAAAA AAAAAAAAAA 2114

SEQ ID NO: 13 Sequence length: 450 Sequence type: Amino acid Topology: Linear Sequence type: Protein sequence Met Val Trp Pro Trp Val Ala Met Ala Ser Arg Trp Gly Pro Leu Ile -36 -35- 30 -25 Gly Leu Ala Pro Cys Cys Leu Trp Leu Leu Gly Ala Val Leu Leu Met -20 -15 -10 -5 Asp Ala Ser Ala Arg Pro Ala Asn His Ser Ser Thr Arg Glu Arg Val 1 5 10 Ala Asn Arg Glu Glu Asn Glu Ile Leu Pro Pro Asp His Leu Asn Gly 15 20 25 Val Lys Leu Glu Met Asp Gly His Leu Asn Arg Gly Phe His Gln Glu 30 35 40 Val Phe Leu Gly Lys Asp Leu Gly Gly Phe Asp Glu Asp Ala Glu Pro 45 50 55 60 Arg Arg Ser Arg Arg Lys Leu Met Val Ile Phe Ser Lys Val Asp Val 65 70 75 Asn Thr Asp Arg Lys Ile Ser Ala Lys Glu Met Gln Arg Trp Ile Met 80 85 90 Glu Lys Thr Ala Glu His Phe Gln Glu Ala Met Glu Glu Ser Lys Thr 95 100 105 His Phe Arg Ala Val Asp Pro Asp Gly Asp Gly His Val Ser Trp Asp 110 115 120 Glu Tyr Lys Val Lys Phe Leu Ala Ser Lys Gly His Ser Glu Lys Glu 125 130 135 140 Val A la Asp Ala Ile Arg Leu Asn Glu Glu Leu Lys Val Asp Glu Glu 145 150 155 Thr Gln Glu Val Leu Glu Asn Leu Lys Asp Arg Trp Tyr Gln Ala Asp 160 165 170 Ser Pro Pro Ala Asp Leu Leu Leu Thr Glu Glu Glu Phe Leu Ser Phe 175 180 185 Leu His Pro Glu His Ser Arg Gly Met Leu Arg Phe Met Val Lys Glu 190 195 200 Ile Val Arg Asp Leu Asp Gln Asp Gly Asp Lys Gln Leu Ser Val Pro 205 210 215 220 Glu Phe Ile Ser Leu Pro Val Gly Thr Val Glu Asn Gln Gln Gly Gln 225 230 235 Asp Ile Asp Asp Asn Trp Val Lys Asp Arg Lys Lys Glu Phe Glu Glu 240 245 250 Leu Ile Asp Ser Asn His Asp Gly Ile Val Thr Ala Glu Glu Leu Glu 255 260 265 Val Ser Pro Gly Ala Ala Val Ser Arg Ser Arg Pro Cys Glu Val Leu 270 275 280 Trp Arg Glu Gly Leu Val Asp Leu Gly Leu Arg Gln Glu Ala Val Leu 285 290 295 300 Val Ser Gly Leu Arg Leu His Leu Gly Trp Ser Leu Gly Arg Leu Leu 305 310 315 Ser Gly Val His Gln Ala Ala Trp Pro Leu Leu Ala Phe Arg Phe Arg 320 325 330 Arg Ile Arg Asp Arg Arg Ala Trp Pro Gly Ser Ile Leu Ser Pro Leu 335 340 345 Arg Arg A rg Gln Gly Ser His Ser Pro Phe Leu Thr Gly Pro Gln Gly 350 355 360 Glu Ala His Ala Gly Pro Ser Met Glu Ala Gly Pro Arg Pro Ser Arg 365 370 375 380 Cys Arg Trp Ala Gly Arg Pro Val Pro His Gly Trp Asn Leu Pro Pro 385 390 395 Ser Ser Glu Ala Phe Ser Leu Val Ser Cys Val Gly Leu Phe Phe Ser 400 405 410 His Cys

SEQ ID NO: 14 Sequence Length: 1353 Sequence Type: Nucleic Acid Number of Strands: Single Strand Topology: Linear Sequence Type: cDNA to mRNA Sequence ATGGTCTGGC CCTGGGTGGC GATGGCGTCC AGGTGGGGTC CCCTCATTGG CCTGGCTCCG 60 TGCTGCCTCT GGCTCCTGGG GGCACGCCTT CTGA GCCTGCCAAC 120 CACTCGTCCA CTCGAGAGAG AGTAGCCAAC AGGGAGGAGA ATGAGATCCT GCCCCCAGAC 180 CACCTGAACG GGGTGAAGCT GGAGATGGAC GGGCACCTCA ATCGCGGCTT CCACCAGGAG 240 GTCTTCCTAG GCAAGGACCT GGGTGGCTTT GATGAGGACG CGGAGCCGCG GCGGAGCCGG 300 AGGAAGCTGA TGGTCATCTT TTCCAAGGTG GATGTGAACA CTGACCGGAA GATCAGTGCC 360 AAGGAGATGC AGCGCTGGAT CATGGAGAAG ACGGCCGAGC ACTTCCAGGA GGCCATGGAG 420 GAGAGCAAGA CACACTTCCG CGCCGTGGAC CCTGACGGGG ACGGTCACGT GTCTTGGGAC 480 GAGTATAAGG TGAAGTTTTT GGCGAGTAAA GGCCATAGCG AGAAGGAGGT TGCCGACGCC 540 ATCAGGCTCA ACGAGGAACT CAAAGTGGAT GAGGAAACAC AGGAAGTCCT GGAGAACCTG 600 AAGGACCGCT GGTACCAGGC GGACAGCCCC CCTGCAGACC TGCTGCTGAC GGAGGAGGAG 660 TTCCTGTCGT TCCTCCACCC CGAGCACAGC CGGGGAATGC TCAGGT TCAT GGTGAAGGAG 720 ATCGTCCGGG ACCTGGACCA GGACGGTGAC AAGCAGCTCT CTGTGCCCGA GTTCATCTCC 780 CTGCCCGTGG GCACCGTGGA GAACCAGCAG GGCCAGGACA TTGACGACAA CTGGGTGAAA 840 GACAGAAAAA AGGAGTTTGA GGAGCTCATT GACTCCAACC ACGACGGCAT CGTGACCGCC 900 GAGGAGCTGG AGGTGAGCCC TGGCGCAGCC GTGTCCCGGA GCCGGCCCTG CGAGGTGCTG 960 TGGCGGGAGG GGCTGGTGGA TCTGGGCCTG AGGCAGGAAG CTGTGCTGGT GTCTGGCCTG 1020 AGACTCCATC TGGGCTGGTC ACTGGGGCGT TTGCTCAGCG GTGTCCACCA GGCTGCATGG 1080 CCGTTGTTGG CGTTTAGGTT CAGACGGATC AGAGACAGGC GAGCCTGGCC GGGCTCCATC 1140 CTCAGCCCCT TGCGGAGGCG TCAGGGTTCT CACAGCCCCT TTTTAACGGG ACCACAAGGG 1200 GAAGCTCATG CTGGGCCCAG CATGGAGGCA GGTCCAAGGC CCAGCAGGTG CAGGTGGGTCGTGCTCTGTGTCTGTTGCGGCGGGCCTG TGCCACATGG CTGGAATTTA CCACCTTCCT TCTCAGCGG

SEQ ID NO: 15 Sequence Length: 2681 Sequence Type: Nucleic Acid Number of Strands: Single Strand Topology: Linear Sequence Type: cDNA to mRNA Sequence CCTGATCCGG GCCCGTTGGCGCGGCGTCACTG ACGCTTCGCT CCGGTCCTCG GATCCCGAGC 60 GCGGGGAGGC AGACCGACTG TGTCCCCATGTCCCCCATGTCCCCCATCATCGA CTGGGGACAC 120 AGAGCCCTCC GTGGTGCCCG GGGATTGGAT TGGAGCCAGG ACCTCACTTC CTCCTCTGCC 180 CCTGCCCCTG CCCCTCCCAG CACCTGGCCC ACACCCTGCA GCCCGCCCCA TGGTCTGGCC 240 CTGGGTGGCG ATGGCGTCCA GGTGGGGTCC CCTCATTGGC CTGGCTCCGT GCTGCCTCTG 300 GCTCCTGGGG GCAGTCCTTC TGATGGACGC GTCTGCACGG CCTGCCAACC ACTCGTCCAC 360 TCGAGAGAGA GTAGCCAACA GGGAGGAGAA TGAGATCCTG CCCCCAGACC ACCTGAACGG 420 GGTGAAGCTG GAGATGGACG GGCACCTCAA TCGCGGCTTC CACCAGGAGG TCTTCCTAGG 480 CAAGGACCTG GGTGGCTTTG ATGAGGACGC GGAGCCGCGG CGGAGCCGGA GGAAGCTGAT 540 GGTCATCTTT TCCAAGGTGG ATGTGAACAC TGACCGGAAG ATCAGTGCCA AGGAGATGCA 600 GCGCTGGATC ATGGAGAAGA CGGCCGAGCA CTTCCAGGAG GCCATGGAGG AGAGCAAGAC 660 ACACTTCCGC GCCGTGGACC CTGACGGGGA CGGTCACGTG TCTTGG GACG AGTATAAGGT 720 GAAGTTTTTG GCGAGTAAAG GCCATAGCGA GAAGGAGGTT GCCGACGCCA TCAGGCTCAA 780 CGAGGAACTC AAAGTGGATG AGGAAACACA GGAAGTCCTG GAGAACCTGA AGGACCGCTG 840 GTACCAGGCG GACAGCCCCC CTGCAGACCT GCTGCTGACG GAGGAGGAGT TCCTGTCGTT 900 CCTCCACCCC GAGCACAGCC GGGGAATGCT CAGGTTCATG GTGAAGGAGA TCGTCCGGGA 960 CCTGGACCAG GACGGTGACA AGCAGCTCTC TGTGCCCGAG TTCATCTCCC TGCCCGTGGG 1020 CACCGTGGAG AACCAGCAGG GCCAGGACAT TGACGACAAC TGGGTGAAAG ACAGAAAAAA 1080 GGAGTTTGAG GAGCTCATTG ACTCCAACCA CGACGGCATC GTGACCGCCG AGGAGCTGGA 1140 GGTGAGCCCT GGCGCAGCCG TGTCCCGGAG CCGGCCCTGC GAGGTGCTGT GGCGGGAGGG 1200 GCTGGTGGAT CTGGGCCTGA GGCAGGAAGC TGTGCTGGTG TCTGGCCTGA GACTCCATCT 1260 GGGCTGGTCA CTGGGGCGTT TGCTCAGCGG TGTCCACCAG GCTGCATGGC CGTTGTTGGC 1320 GTTTAGGTTC AGACGGATCA GAGACAGGCG AGCCTGGCCG GGCTCCATCC TCAGCCCCTT 1380 GCGGAGGCGT CAGGGTTCTC ACAGCCCCTT TTTAACGGGA CCACAAGGGG AAGCTCATGC 1440 TGGGCCCAGC ATGGAGGCAG GTCCAAGGCC CAGCAGGTGC AGGTGGGCGG GGCGGCCTGT 1500 GCCACATGGC TGGAATTTAC CACCTTCCTC TGAAGCGTTT TCACTGGTAT CATGTG TAGG 1560 CTTGTTTTTC TCCCACTGCT GAGTGAGTCA TCTTGTTTTT ATGTAGAATC CTGTGATTCC 1620 TGGCGACAGC CAGTGGGCCC GGCCCAGGTT AGGGATCCTT CAGAACTGGG GTCCAGGCCT 1680 GTGTAGCCCC TGTGCCCCGT TACCCCTGCT GCCCCCGGGC AGGCCTTCCG GGGCCACCGG 1740 CTTCTCCCTG CCCTGTGTTT TAATTTGTCC CGCCTCCCTT CGGGAACCTT CCAGAACGTG 1800 CCCACACTCC CGCTGCAGCC AATAGGCACC TTAAATAGCC ACTTCGTGCG GCTGGCCGCG 1860 GAGCTCGGAG GGGGAAAGGC GACGCTGACC TGTGCCCCGC TCGCCCGCAG AGCTACATGG 1920 ACCCCATGAA CGAGTACAAC GCGCTGAACG AGGCCAAGCA GATGATCGCC GTCGCCGACG 1980 AGAACCAGAA CCACCACCTG GAGCCCGAGG AGGTGCTCAA GTACAGCGAG TTCTTCACGG 2040 GCAGCAAGCT GGTGGACTAC GCGCGCAGCG TGCACGAGGA GTTTTGAGCG CCCGGCCGCG 2100 CCCCGCGCCG CCCCCCACGC ACCACCGGGG GGGCCTCGCG GGTGACTCCG GCCTCCGTGG 2160 CTGTCCCGGA CCCCACCTCT TCTCTGCCGC CCACCACCGG CCGACCGACC GCGGCTTCCC 2220 CAGTTGATGA GCGGCGTGTC CCCTCTGCAG CGCGCACCCC GGCGGGGCTT TGGCTGTGAC 2280 GCGGTCGGGG CGCGGGGCTG GTCTGTGGCC CCGCGGCGCG CCTCCTCCCT GGTCCCTCGA 2340 AATCGTGGCA TCTCACTTCT GAGAACGAAA TCTCGCTTCA GTCACTCTGC CGAAGGCGCT 2 400 GACGGCATCG CGGCCGGAAC CTCTGGGCCC GGCCCCTCCC AGGGCCGCCG CTCCGTGGGA 2460 AAAAACAGCT CCTCCATTTC CTTGGAAACT GAACGATTAT TAAAAATAGA TAAACTTCGC 2520 TGGAAATGAG TAGCCAGGAA GTTCAGGGGA GGGTCCGGGG TCCTTCCCGG GGCCTGGCGT 2580 GTCGGAACCA CCCAGGTCCC GCAGCTGCCT CTGAGAAAAT CCAAATATTT TTTGTGACAA 2640 GAATCACAAA CATTTACTTT AAATATAAAA AAAAAAAAAA A 2681

SEQ ID NO: 16 Sequence length: 2681 Sequence type: Nucleic acid Number of strands: Single strand Topology: Linear Sequence type: cDNA to mRNA Origin: Organism name: Homo Sapiens Cell line: T98G Sequence Feature Symbol representing the feature: CDS Location: 230..1582 Method by which the feature was determined: P Feature symbol: sig_peptide Location: 230..337 Method by which the feature was determined: S Symbol representing feature: mat_peptide Location: 338..1582 method to determine the characteristics: S sequence CCTGATCCGG GCCCGTTGGC GGCGTCACTG ACGCTTCGCT CCGGTCCTCG GATCCCGAGC 60 GCGGGGAGGC AGACCGACTG TGAGCTGCTT GTCCCCATCC TGCGGACGTC CTGGGGACAC 120 AGAGCCCTCC GTGGTGCCCG GGGATTGGAT TGGAGCCAGG ACCTCACTTC CTCCTCTGCC 180 CCTGCCCCTG CCCCTCCCAG CACCTGGCCC ACACCCTGCA GCCCGCCCC ATG GTC 235 Met Val -36 -35 TGG CCC TGG GTG GCG ATG GCG TCC AGG TGG GGT CCC CTC ATT GGC CTG 283 Trp Pro Trp Val Ala Met Ala Ser Arg Trp Gly Pro Leu Ile Gly Leu -30 -25 -20 GCT CCG TGC TGC CTC TGG CTC CTG GGG GCA GTC CTT CTG ATG GAC GCG 331 Ala Pro Cys Cys Leu Trp Leu Leu Gly Ala Val Leu Leu Met Asp Ala -15 -10 -5 TCT GCA CGG CCT GCC AAC CAC TCG TCC ACT CGA GAG AGA GTA GCC AAC 379 Ser Ala Arg Pro Ala Asn His Ser Ser Thr Arg Glu Arg Val Ala Asn 1 5 10 AGG GAG GAG AAT GAG ATC CTG CCC CCA GAC CAC CTG AAC GGG GTG AAG 427 Arg Glu Glu Asn Glu Ile Leu Pro Pro Asp His Leu Asn Gly Val Lys 15 20 25 30 CTG GAG ATG GAC GGG CAC CTC AAT CGC GGC TTC CAC CAG GAG GTC TTC 475 Leu Glu Met Asp Gly His Leu Asn Arg Gly Phe His Gln Glu Val Phe 35 40 45 CTA GGC AAG GAC CTG GGT GGC TTT GAT GAG GAC GCG GAG CCG CGG CGG 523 Leu Gly Lys Asp Leu Gly Gly Phe Asp Glu Asp Ala Glu Pro Arg Arg 50 55 60 AGC CGG AGG AAG CTG ATG GTC ATC TTT TCC AAG GTG GAT GTG AAC ACT 571 Ser Arg Arg Lys Leu Met Val Ile Phe Ser Lys Val Asp Val Asn Thr 65 70 75 GAC CGG AAG ATC AGT GCC AAG GAG ATG CAG CGC TGG ATC ATG GAG AAG 619 Asp Arg Lys Ile Ser Ala Lys Glu Met Gln Arg Trp Ile Met Glu Lys 80 85 90 ACG GCC GAG CAC TTC CAG GAG GCC ATG GAG GAG AGC AAG ACA CAC TTC 667 Thr Ala Glu His Phe Gln Glu Ala Met Glu Glu Ser Lys Thr His Phe 95 100 105 110 CGC GCC GTG GAC CCT GAC GGG GAC GGT CAC GTG TCT TGG GAC GAG TAT 715 Arg Ala Val Asp Pro Asp Gly Asp Gly His Val Ser Trp Asp Glu Tyr 115 120 125 AAG GTG AAG TTT TTG GCG AGT AAA GGC CAT AGC GAG AAG GAG GTT GCC 763 Lys Val Lys Phe Leu Ala Ser Lys Gly His Ser Glu Lys Glu Val Ala 130 135 140 GAC GCC ATC AGG CTC AAC GAG GAA CTC AAA GTG GAT GAG GAA ACA CAG 811 Asp Ala Ile Arg Leu Asn Glu Glu Leu Lys Val Asp Glu Glu Thr Gln 145 150 155 GAA GTC CTG GAG AAC CTG AAG GAC CGC TGG TAC CAG GCG GAC AGC CCC 859 Glu Val Leu Glu Asn Leu Lys Asp Arg Trp Tyr Gln Ala Asp Ser Pro 160 165 170 CCT GCA GAC CTG CTG CTG ACG GAG GAG GAG TTC CTG TCG TTC CTC CAC 907 Pro Ala Asp Leu Leu Leu Leu Thr Glu Glu Glu Phe Leu Ser Phe Leu His 175 180 185 190 CCC GAG CAC AGC CGG GGA ATG CTC AGG TTC ATG GTG AAG GAG ATC GTC 955 Pro Glu His Ser Arg Gly Met Leu Arg Phe Met Val Lys Glu Ile Val 195 200 205 CGG G AC CTG GAC CAG GAC GGT GAC AAG CAG CTC TCT GTG CCC GAG TTC 1003 Arg Asp Leu Asp Gln Asp Gly Asp Lys Gln Leu Ser Val Pro Glu Phe 210 215 220 ATC TCC CTG CCC GTG GGC ACC GTG GAG AAC CAG CAG GGC CAG GAC ATT 1051 Ile Ser Leu Pro Val Gly Thr Val Glu Asn Gln Gln Gly Gln Asp Ile 225 230 235 GAC GAC AAC TGG GTG AAA GAC AGA AAA AAG GAG TTT GAG GAG CTC ATT 1099 Asp Asp Asn Trp Val Lys Asp Arg Lys Lys Glu Phe Glu Glu Leu Ile 240 245 250 GAC TCC AAC CAC GAC GGC ATC GTG ACC GCC GAG GAG CTG GAG GTG AGC 1147 Asp Ser Asn His Asp Gly Ile Val Thr Ala Glu Glu Leu Glu Val Ser 255 260 265 270 CCT GGC GCA GCC GTG TCC CGG AGC CGG CCC TGC GAG GTG CTG TGG CGG 1195 Pro Gly Ala Ala Val Ser Arg Ser Arg Pro Cys Glu Val Leu Trp Arg 275 280 285 GAG GGG CTG GTG GAT CTG GGC CTG AGG CAG GAA GCT GTG CTG GTG TCT 1243 Glu Gly Leu Val Asp Leu Gly Leu Arg Gln Glu Ala Val Leu Val Ser 290 295 300 GGC CTG AGA CTC CAT CTG GGC TGG TCA CTG GGG CGT TTG CTC AGC GGT 1291 Gly Leu Arg Leu His Leu Gly Trp Ser Leu Gly Arg Leu Leu Ser G ly 305 310 315 GTC CAC CAG GCT GCA TGG CCG TTG TTG GCG TTT AGG TTC AGA CGG ATC 1339 Val His Gln Ala Ala Trp Pro Leu Leu Ala Phe Arg Phe Arg Arg Ile 320 325 330 AGA GAC AGG CGA GCC TGG CCG GGC TCC ATC CTC AGC CCC TTG CGG AGG 1387 Arg Asp Arg Arg Ala Trp Pro Gly Ser Ile Leu Ser Pro Leu Arg Arg 335 340 345 350 CGT CAG GGT TCT CAC AGC CCC TTT TTA ACG GGA CCA CAA GGG GAA GCT 1435 Arg Gln Gly Ser His Ser Pro Phe Leu Thr Gly Pro Gln Gly Glu Ala 355 360 365 CAT GCT GGG CCC AGC ATG GAG GCA GGT CCA AGG CCC AGC AGG TGC AGG 1483 His Ala Gly Pro Ser Met Glu Ala Gly Pro Arg Pro Ser Arg Cys Arg 370 375 380 TGG GCG GGG CGG CCT GTG CCA CAT GGC TGG AAT TTA CCA CCT TCC TCT 1531 Trp Ala Gly Arg Pro Val Pro His Gly Trp Asn Leu Pro Pro Ser Ser 385 390 395 GAA GCG TTT TCA CTG GTA TCA TGT GTA GGC TTG TTT TTC TCC CAC TGC 1579 Glu Ala Phe Ser Leu Val Ser Cys Val Gly Leu Phe Phe Ser His Cys 400 405 410 TGA GTGAGTCATC TTGTTTTTAT GTAGAATCCT GTGATTCCTG GCGACAGCCA 1632 * 415 GTGGGCCCGG CCCAGGTTAG GGATCCTTCA GAACTGGG GT CCAGGCCTGT GTAGCCCCTG 1692 TGCCCCGTTA CCCCTGCTGC CCCCGGGCAG GCCTTCCGGG GCCACCGGCT TCTCCCTGCC 1752 CTGTGTTTTA ATTTGTCCCG CCTCCCTTCG GGAACCTTCC AGAACGTGCC CACACTCCCG 1812 CTGCAGCCAA TAGGCACCTT AAATAGCCAC TTCGTGCGGC TGGCCGCGGA GCTCGGAGGG 1872 GGAAAGGCGA CGCTGACCTG TGCCCCGCTC GCCCGCAGAG CTACATGGAC CCCATGAACG 1932 AGTACAACGC GCTGAACGAG GCCAAGCAGA TGATCGCCGT CGCCGACGAG AACCAGAACC 1992 ACCACCTGGA GCCCGAGGAG GTGCTCAAGT ACAGCGAGTT CTTCACGGGC AGCAAGCTGG 2052 TGGACTACGC GCGCAGCGTG CACGAGGAGT TTTGAGCGCC CGGCCGCGCC CCGCGCCGCC 2112 CCCCACGCAC CACCGGGGGG GCCTCGCGGG TGACTCCGGC CTCCGTGGCT GTCCCGGACC 2172 CCACCTCTTC TCTGCCGCCC ACCACCGGCC GACCGACCGC GGCTTCCCCA GTTGATGAGC 2232 GGCGTGTCCC CTCTGCAGCG CGCACCCCGG CGGGGCTTTG GCTGTGACGC GGTCGGGGCG 2292 CGGGGCTGGT CTGTGGCCCC GCGGCGCGCC TCCTCCCTGG TCCCTCGAAA TCGTGGCATC 2352 TCACTTCTGA GAACGAAATC TCGCTTCAGT CACTCTGCCG AAGGCGCTGA CGGCATCGCG 2412 GCCGGAACCT CTGGGCCCGG CCCCTCCCAG GGCCGCCGCT CCGTGGGAAA AAACAGCTCC 2472 TCCATTTCCT TGGAAACTGA ACGATTATTA AAAATAGATA AAC TTCGCTG GAAATGAGTA 2532 GCCAGGAAGT TCAGGGGAGG GTCCGGGGTC CTTCCCGGGG CCTGGCGTGT CGGAACCACC 2592 CAGGTCCCGC AGCTGCCTCT GAGAAAATCC AAATATTTTT TGTGACAAGA ATCACAAACA 2652 TTTACTTTAA ATATAAAAAA AAAAAAAAA 2681

[Brief description of drawings]

FIG. 1 is a conceptual diagram of a method for preparing a cDNA library of the present invention.

FIG. 2 is a construction diagram of a plasmid vector pUCSRαML2.

FIG. 3 is a construction diagram of pUCSRαML2-hTac.

FIG. 4 Mouse type ST-NT3 and human type ST-NT3
Shows a comparison of α, 3β and 3γ sequences.

─────────────────────────────────────────────────── ─── Continuation of the front page (51) Int.Cl. ⁶ Identification code Internal reference number FI Technical display location A61K 39/395 8828-4B C12N 1/21 C12P 21/02 C C07H 21/04 21/08 C07K 16 / 18 A61K 48/00 C12N 1/21 37/02 AAA 15/09 ZNA ABA C12P 21/02 ABJ 21/08 ABY // A61K 48/00 9162-4B C12N 15/00 ZNAA (C12N 1/21 C12R 1: 19) (C12P 21/02 C12R 1:19) (72) Inventor Hideaki Tada 3-1-1 Sakurai, Shimamoto-cho, Mishima-gun, Osaka Prefecture Minase Research Laboratory, Ono Pharmaceutical Co., Ltd. (72) Daikichi Fukushima Mishima, Osaka Prefecture 3-1-1, Sakurai, Korishimamoto-cho Minase Research Laboratory, Ono Pharmaceutical Co., Ltd. (54) [Title of Invention] Novel polypeptide, its production method, its Which encodes the polypeptide, a vector comprising the DNA, a host cell transformed with the vector, an antibody of the polypeptide, and a pharmaceutical composition containing the peptide or antibody.

Claims (20)

[Claims] 1. A polypeptide comprising the amino acid sequence represented by SEQ ID NO: 1, which is in a substantially pure form, a homologue thereof, a fragment thereof or a homologue of a fragment thereof. 2. The polypeptide according to claim 1, which consists of the amino acid sequence represented by SEQ ID NO: 1. 3. A DNA encoding the polypeptide according to claim 2. 4. The DNA according to claim 3, which has the base sequence represented by SEQ ID NO: 2, or a fragment which selectively hybridizes to the sequence. 5. The DNA according to claim 3, which has the nucleotide sequence represented by SEQ ID NO: 3, or a fragment which selectively hybridizes to the sequence. 6. A replication or expression vector comprising the DNA according to any one of claims 3 to 5. 7. A host cell transformed with the replication or expression vector of claim 6. 8. A method for producing the polypeptide according to claim 1, which comprises culturing the host cell according to claim 7 under conditions for expressing the polypeptide according to claim 1 or 2. 9. A monoclonal or polyclonal antibody of the polypeptide according to claim 1 or 2. 10. A pharmaceutical composition comprising the polypeptide according to claim 1 or 2 or the antibody according to claim 9 and a pharmaceutically acceptable excipient and / or carrier. 11. SEQ ID NO: 5 in substantially pure form,
A polypeptide comprising the amino acid sequence represented by 9 or 13, a homologue thereof, a fragment thereof or a homologue of the fragment. 12. The polypeptide according to claim 11, which consists of the amino acid sequence shown in SEQ ID NO: 5, 9 or 13. 13. A DNA encoding the polypeptide according to claim 11. 14. The DNA according to claim 13, which has the nucleotide sequence represented by SEQ ID NO: 6, 10 or 14, or a fragment which selectively hybridizes to the sequence. 15. The DNA according to claim 13, which has the nucleotide sequence represented by SEQ ID NO: 7, 11 or 15, or a fragment which selectively hybridizes to the sequence. 16. A replication or expression vector comprising the DNA according to any one of claims 13 to 15. 17. A host cell transformed with the replication or expression vector of claim 16. 18. A method for producing the polypeptide according to claim 17, which comprises culturing the host cell according to claim 17 under conditions for expressing the polypeptide according to claim 11 or 12. 19. A monoclonal or polyclonal antibody of the polypeptide according to claim 11 or 12. 20. A pharmaceutical composition comprising the polypeptide according to claim 11 or 12 or the antibody according to claim 19 and a pharmaceutically acceptable excipient and / or carrier.