KR20060107489A - Human protooncogene trg and protein encoded therein - Google Patents

Abstract

The present invention relates to a primary cancer gene and a protein encoded by the primary cancer gene, wherein the primary cancer gene of the present invention is involved in human cancer and can be effectively used for diagnosis of cancer including uterine cancer, leukemia, lymphoma, colorectal cancer, lung cancer and skin cancer. have.

Description

HUMAN PROTOONCOGENE TRG AND PROTEIN ENCODED THEREIN}

1 shows H20-121 DNA fragment (a) in normal cervical tissue, cervical tumor tissue, metastatic lymph node tumor tissue and CUMC-6 cancer cells; H93-811 DNA fragment (b); H117-321 DNA fragment (c); H38-211 DNA fragment (d); H38-621 DNA fragment (e); H96 DNA fragment (f); H94 DNA fragment (g); H42 DNA fragment (h); H109 DNA fragment (i); H119 DNA fragment (j); H201 DNA fragment (k); H151 DNA fragment (l); H132 DNA fragment (m); H141 DNA fragment (n); H181 DNA fragment (o); H134 It shows the result of differential development reverse transcription polymerase chain reaction (DDRT-PCR) confirming the expression of DNA fragment (p).

2 shows TRG3 (top a) of the present invention in normal cervical tissue, cervical cancer tissue, cervical metastatic lymph node tissue and cervical cancer cell lines; TRG4 (top b); TRG5 (c top); TRG6 (d top); TRG7 (top e); TRG9 (f top); TRG10 (g top); TRG11 (h top); TRG12 (top i) TRG13 (top j); TRG14 (k top); TRG15 (l top); TRG16 (top m); TRG17 (n top); TRG18 (top top); TRB20 (p top) shows the results of Northern blot analysis confirming the expression of the primary cancer gene, the bottom of each Figure 2 shows the results obtained by hybridizing the same samples as the top of each Figure 2 with β-actin,

Figure 3a shows the results of the Northern blot analysis confirming the expression of the TRG3 primary cancer gene of the present invention in normal human 12-row multiple tissues, Figure 3b shows the results obtained by hybridizing the same sample as in Figure 3a with β-actin ;

Figure 4a shows the results of Northern blot analysis confirming the expression of the TRG4 primary cancer gene of the present invention in normal human 12-row multiple tissue, Figure 4b shows the results obtained by hybridizing the same sample as β-actin 4a ,

Figure 5a shows the results of Northern blot analysis confirming the expression of the TRG5 primary cancer gene of the present invention in normal human 12-row multiple tissues, Figure 5b shows the results obtained by hybridizing the same sample as β-actin 5a ,

Figure 6a shows the results of Northern blot analysis confirming the expression of the TRG6 primary cancer gene of the present invention in normal human 12-row multiple tissues, Figure 6b shows the results obtained by hybridizing the same sample as β-actin 6a ;

Figure 7a shows the results of Northern blot analysis confirming the expression of the TRG7 primary cancer gene of the present invention in normal human 12-row multiple tissues, Figure 7b shows the results obtained by hybridizing the same sample as β-actin 7a ,

Figure 8a shows the results of Northern blot analysis confirming the expression of the TRG9 primary cancer gene of the present invention in normal human 12-row multiple tissues, Figure 8b shows the result obtained by hybridizing the same sample as β-actin 8a ,

Figure 9a shows the results of Northern blot analysis confirming the expression of the TRG10 primary cancer gene of the present invention in normal human 12-row multiple tissues, Figure 9b shows the results obtained by hybridizing the same sample as β-actin 9a ;

Figure 10a shows the results of Northern blot analysis confirming the expression of the TRG11 primary cancer gene of the present invention in normal human 12-row multiple tissues, Figure 10b shows the results obtained by hybridizing the same sample as β-actin 10a ,

Figure 11a shows the results of Northern blot analysis confirming the expression of the TRG12 primary cancer gene of the present invention in normal human 12-row multiple tissues, Figure 11b shows the results obtained by hybridizing the same sample as β-actin 11a ,

Figure 12a shows the results of Northern blot analysis confirming the expression of the TRG13 primary cancer gene of the present invention in normal human 12-row multiple tissues, Figure 12b shows the results obtained by hybridizing the same sample as β-actin with Figure 12a ;

Figure 13a shows the results of the Northern blot analysis confirming the expression of the TRG14 primary cancer gene of the present invention in normal human 12-row multiple tissues, Figure 13b shows the results obtained by hybridizing the same sample as β-actin 13a ,

Figure 14a shows the results of the Northern blot analysis confirming the expression of the TRG15 primary cancer gene of the present invention in normal human 12-row multiple tissues, Figure 14b shows the results obtained by hybridizing the same sample as β-actin 14a ,

Figure 15a shows the results of Northern blot analysis confirming the expression of the TRG16 primary cancer gene of the present invention in normal human 12-row multiple tissues, Figure 15b shows the results obtained by hybridizing the same sample as β-actin 15a ;

Figure 16a shows the results of the Northern blot analysis confirming the expression of the TRG17 primary cancer gene of the present invention in normal human 12-row multiple tissue, Figure 16b shows the results obtained by hybridizing the same sample as β-actin 16a ,

Figure 17a shows the results of Northern blot analysis confirming the expression of the TRG18 primary cancer gene of the present invention in normal human 12-row multiple tissues, Figure 17b shows the results obtained by hybridizing the same sample as β-actin Figure 17a ,

Figure 18a shows the results of Northern blot analysis confirming the expression of the TRG20 proto-oncogene of the present invention in normal human 12-row multiple tissues, Figure 18b shows the results obtained by hybridizing the same sample as β-actin in Figure 18a .

Figure 19a shows the results of Northern blot analysis confirming the expression of the TRG3 primary oncogene of the present invention in human cancer cell lines, Figure 19b shows the results obtained by hybridizing the same sample as in Figure 19a with β-actin,

Figure 20a shows the results of Northern blot analysis confirming the expression of the TRG4 primary oncogenes in human cancer cell lines, Figure 20b shows the results obtained by hybridizing the same sample as in Figure 20a with β-actin,

Figure 21a shows the results of Northern blot analysis confirming the expression of the TRG5 primary cancer gene of the present invention in human cancer cell lines, Figure 21b shows the result obtained by hybridizing the same sample as β-actin, Figure 21a,

Figure 22a shows the results of Northern blot analysis confirming the expression of the TRG6 primary oncogene of the present invention in human cancer cell lines, Figure 22b shows the results obtained by hybridizing the same sample as in Figure 22a with β-actin;

Figure 23a shows the results of Northern blot analysis confirming the expression of the TRG7 primary oncogene of the present invention in human cancer cell lines, Figure 23b shows the result obtained by hybridizing the same sample as in Figure 23a with β-actin,

Figure 24a shows the results of Northern blot analysis confirming the expression of the TRG9 primary cancer gene of the present invention in human cancer cell lines, Figure 24b shows the result obtained by hybridizing the same sample as in Figure 24a with β-actin,

Figure 25a shows the results of Northern blot analysis confirming the expression of the TRG10 primary cancer gene of the present invention in human cancer cell lines, Figure 25b shows the results obtained by hybridizing the same sample as in Figure 25a with β-actin;

Figure 26a shows the results of Northern blot analysis confirming the expression of the TRG11 primary cancer gene of the present invention in human cancer cell lines, Figure 26b shows the results obtained by hybridizing the same sample as in Figure 26a with β-actin,

Figure 27a shows the results of Northern blot analysis confirming the expression of the TRG12 proto-oncogene of the present invention in human cancer cell lines, Figure 27b shows the results obtained by hybridizing the same sample as in Figure 27a with β-actin,

FIG. 28a shows the results of Northern blot analysis confirming the expression of the TRG13 proto-oncogene of the present invention in human cancer cell lines, and FIG. 28b shows the result obtained by hybridizing the same sample as in FIG. 28a with β-actin; FIG.

FIG. 29A shows a result of Northern blot analysis confirming the expression of TRG14 proto-oncogene of the present invention in human cancer cell lines, and FIG. 29B shows the result obtained by hybridizing the same sample as that of FIG. 29A with β-actin. FIG.

FIG. 30A shows a result of Northern blot analysis confirming the expression of TRG15 proto-oncogene of the present invention in human cancer cell lines, and FIG. 30B shows the result obtained by hybridizing the same sample as that of FIG. 30A with β-actin. FIG.

Figure 31a shows the results of Northern blot analysis confirming the expression of the TRG16 primary oncogene of the present invention in human cancer cell lines, Figure 31b shows the results obtained by hybridizing the same sample as in Figure 31a with β-actin;

32a shows the results of Northern blot analysis confirming the expression of the TRG17 proto-oncogene of the present invention in human cancer cell lines, and FIG. 32b shows the result obtained by hybridizing the same sample as in FIG. 32a with β-actin. FIG.

Figure 33a shows the results of Northern blot analysis confirming the expression of the TRG18 primary oncogene of the present invention in human cancer cell lines, Figure 33b shows the results obtained by hybridizing the same sample as in Figure 33a with β-actin,

Figure 34a shows the results of Northern blot analysis confirming the expression of the TRG20 primary cancer gene of the present invention in human cancer cell lines, Figure 34b shows the result obtained by hybridizing the same sample as β-actin 34a.

35 shows TRG3 (a) of the present invention; TRG4 (b); TRG5 (c); TRG6 (d); TRG7 (e); TRG9 (f); TRG10 (g); TRG11 (h); TRG12 (i) TRG13 (j); TRG14 (k); TRG15 (l); TRG16 (m); TRG17 (n); TRG18 (o); After transduction of TRG20 (p) prokaryotic gene into E. coli, the size of protein expressed before and after L-Arabinose induction was electrophoresed on sodium dodecyl sulfate-polyacrylamide gel (SDS-PAGE). The result is.

The present invention relates to novel primary oncogenes and proteins encoded by them that exhibit cancer development and cancer metastasis.

Higher animals, including humans, have about 30,000 genes, but only about 15% of them are expressed in each individual. Thus, which gene is selected and expressed determines all phenomena of life: development, differentiation, homeostasis, response to stimuli, cell division cycle regulation, aging and apoptosis (program cell death). (Liang, P. and AB Pardee, Science 257: 967-971, 1992).

Pathological phenomena, such as tumorigenesis, are caused by genetic mutations that eventually lead to changes in gene expression. Thus, comparison of gene expressions between different cells is a fundamental and effective approach to understanding various biological phenomena. For example, Liang, P. and AB Pardee, Science 257: 967-971, 1992) suggested that the differential display method of mRNA is currently used to search for tumor suppressor genes, cell cycle-related genes, and apoptosis-related transcriptional regulatory genes. It is effectively used, and is also used in a variety of ways to correlate various genes in a single cell.

Taken together, studies on tumor development suggest that several genetic changes, such as loss of chromosomal heterozygosity, activation of proto-oncogenes, and inactivation of other tumor suppressor genes, including the p53 gene, may be present in tumor tissues. Accumulate and cause human tumors (Bishop, JM, Cell) 64: 235-248, 1991; Hunter, T., Cell 64: 249-270, 1991). In addition, 10 to 30% of cancers are reported to be activated by the amplification of the primary cancer gene, activation of the primary cancer gene plays an important role in the etiology of many cancers, it is required to reveal this.

Accordingly, the inventors of the present invention have approached the developmental mechanism of cervical cancer at the level of the primary cancer gene, and found that the primary cancer gene named human transformation-related gene (TRG) is specifically increased only in cancer cells. The primary cancer gene can be effectively used for the diagnosis, prevention and treatment of various cancers such as uterine cancer, leukemia, lymphoma, colon cancer, lung cancer and skin cancer.

It is an object of the present invention to provide novel far-oncogenes and fragments thereof.

Another object of the present invention is to provide a recombinant vector comprising the above-described oncogene or a fragment thereof and a microorganism transformed thereby.

Still another object of the present invention is to provide a protein and fragment thereof encoded by the primary cancer gene.

Still another object of the present invention is to provide a kit for diagnosing cancer, comprising the original oncogene or a fragment thereof.

Still another object of the present invention is to provide a kit for diagnosing cancer comprising the protein or fragment thereof.

According to the above object, in the present invention, SEQ ID NO: 1; SEQ ID NO: 5; SEQ ID NO: 9; SEQ ID NO: 13; SEQ ID NO: 17; SEQ ID NO: 21; SEQ ID NO: 25; SEQ ID NO: 29; SEQ ID NO: 33; SEQ ID NO: 37; SEQ ID NO: 41; SEQ ID NO: 45; SEQ ID NO: 49; SEQ ID NO: 53; SEQ ID NO: 57; Or a primary cancer gene or fragment thereof having the nucleotide sequence of SEQ ID NO: 61.

According to the above another object, the present invention provides a recombinant vector and the microorganism transformed by the above-described oncogene or a fragment thereof.

According to another object of the present invention, SEQ ID NO: 2 encoded by the primary cancer gene; SEQ ID NO: 6; SEQ ID NO: 10; SEQ ID NO: 14; SEQ ID NO: 18; SEQ ID NO: 22; SEQ ID NO: 26; SEQ ID NO: 30; SEQ ID NO: 34; SEQ ID NO: 38; SEQ ID NO: 42; SEQ ID NO: 46; SEQ ID NO: 50; SEQ ID NO: 54; SEQ ID NO: 58; Or a protein having an amino acid sequence of SEQ ID NO: 62 or a fragment thereof.

In accordance with another object, the present invention provides a kit for diagnosing cancer, including the primary cancer gene or a fragment thereof.

In accordance with another object, the present invention provides a kit for diagnosing cancer, comprising the original cancer protein or fragment thereof.

According to another object of the present invention, the present invention has a base sequence complementary to all or part of the mRNA sequence transcribed from the primary cancer gene or fragment thereof, and binds to the mRNA to inhibit the expression of the primary cancer gene or fragment. Provide sense genes.

In accordance with another object, the present invention provides an anticancer and anti-transition composition comprising the anti-sense gene as an active ingredient.

Hereinafter, the present invention will be described in detail.

One. TRG  3

Human transformation-related gene 3 (hereinafter referred to as TRG3), which is a protocogene of the present invention, has an entire sequence of 1,703 bp in length as set forth in SEQ ID NO: 1.

In the nucleotide sequence of SEQ ID NO: 1, the open reading frame corresponding to the nucleotide number 113 to 1522 site (1520-1522: end codon) is the entire protein coding region and the amino acid sequence derived therefrom is SEQ ID NO: It is shown in 2 and consists of 469 amino acids (hereinafter referred to as "TRG3 protein").

The base sequence of SEQ ID NO: 1 was registered in the NIH GenBank database of the National Institutes of Health as registration number AY189688 (published: April 8, 2005). Homo sapiens CGI-51 protein gene and gene sequence were confirmed to be similar. However, this study identified TRG3 proto-oncogenes, which are highly expressed in various human tumors, including uterine cancer, and very low in various normal tissues.

However, due to the degeneracy of the codon or in consideration of the preferred codon in the organism to express the primary cancer gene, the primary cancer gene of the present invention does not change the amino acid sequence of the carcinogenic protein expressed from the coding region. In the coding region, various modifications may be made, and various modifications or modifications may be made within a range that does not affect the expression of genes in portions other than the coding region, and such modified genes are also included in the scope of the present invention. Accordingly, the present invention also encompasses polynucleotides and fragments of these genes having base sequences substantially the same as the original oncogene of SEQ ID NO: 1. Substantially identical polynucleotides are DNAs encoding a protein translation product identical to SEQ ID NO: 2, meaning those having sequence homology of at least 80%, preferably at least 90%, most preferably at least 95% with SEQ ID NO: 1 do.

The protein expressed from the proto-oncogene of the present invention consists of 469 amino acids, has an amino acid sequence set forth in SEQ ID NO: 2, and is about 52 kDa in size. However, even in the amino acid sequence of the protein, substitution, addition or deletion of amino acids may be made within a range that does not affect the function of the protein, and only a part of the protein may be used depending on the purpose. Such modified amino acid sequences are also within the scope of the present invention. Accordingly, the present invention also encompasses polypeptides and fragments thereof having substantially the same amino acid sequence as the carcinogenic protein, wherein substantially identical polypeptides comprise at least 80%, preferably at least 90% and most preferably at least 95% sequences Means those with homology.

2 . TRG 4

The human gene transformation-related gene 4 (hereinafter referred to as TRG4), which is a protocogene of the present invention, has a total base sequence of 2,576 bp in length as described in SEQ ID NO: 5.

In the nucleotide sequence of SEQ ID NO: 5, the open reading frame corresponding to the nucleotides nucleotides 87 to 482 (480-482: end codon) is the entire protein coding region and the amino acid sequence derived from it is the sequence number: It is shown in 6 and consists of 131 amino acids (hereinafter referred to as "TRG4" protein).

The base sequence of SEQ ID NO: 5 is registered in the US National Institutes of Health's NIH GenBank database with registration number AY189690 (published date: April 8, 2005) and is registered as NM_022463 in this database. Homo sapiens nucleoredoxin (NXN) genes and gene bases have been identified to be similar, but protein sequences are completely different genes. However, this study has led to the discovery of TRG4 primary oncogenes, which have high expression in many human tumors, including uterine cancer, and very low expression in normal tissues.

However, due to the degeneracy of codons or in view of the preferred codons in organisms that wish to express the above-described source cancer genes, the source cancer genes of the present invention do not change the amino acid sequence of the oncogenic proteins expressed from the coding region. Many variations and modifications of the coding region may occur, and the coding region may not affect the expression of the gene, except in the coding region, and such modifications may also be included in the scope of the present invention. Thus, the present invention also encompasses polynucleotides having substantially the same base sequence as the original cancer gene of SEQ ID NO: 5 and fragments of the gene. Substantially identical polynucleotides are DNAs encoding the same protein translation product as SEQ ID NO: 6, which means that they have sequence homology of at least 80%, preferably at least 90% and most preferably at least 95%, of sequence identity with SEQ ID NO: 5. do.

The protein expressed from the original cancer gene of the present invention is composed of 131 amino acids and has the amino acid sequence described in SEQ ID NO: 6, and has a size of about 14 kDa. However, even in the amino acid sequences of the above proteins, amino acid substitutions, additions or deletions may be made within a range that does not affect the function of the protein. Depending on the purpose, only a part of the protein may be used. Such modified “amino acid sequences” are also included within the scope of the present invention. Thus, the present invention also encompasses polypeptides and fragments thereof having substantially the same amino acid sequence as the carcinogenic proteins, with substantially identical polypeptides of 80% or more, preferably 90% or more, and most preferably 95% or more. It means things with same-sex.

3. TRG  5

Human transformation-related gene 5 (hereinafter referred to as TRG5), which is the protocogene of the present invention, has an entire sequence of 1,334 bp in length as set forth in SEQ ID NO: 9.

In the nucleotide sequence of SEQ ID NO: 9, the open reading frame corresponding to the nucleotide number 88 to 1092 region (1090-1092: end codon) is the entire protein coding region, and the amino acid sequence derived therefrom is SEQ ID NO: It is shown at 10 and consists of 334 amino acids (hereinafter referred to as "TRG5 protein").

The base sequence of SEQ ID NO: 9 was registered in the NIH GenBank database of the National Institutes of Health as No. AY189689 (published: April 8, 2005). As a result of sequencing analysis, the sequence of NM_002300 is registered in this database. Homo sapiens lactate dehydrogenase B (LDHB) gene and gene sequence were confirmed to be similar. However, this study identified TRG5 proto-oncogenes, which are highly expressed in various human tumors, including uterine cancer, and very low in various normal tissues.

However, due to the degeneracy of the codon or in consideration of the preferred codon in the organism to express the primary cancer gene, the primary cancer gene of the present invention does not change the amino acid sequence of the carcinogenic protein expressed from the coding region. In the coding region, various modifications may be made, and various modifications or modifications may be made within a range that does not affect the expression of genes in portions other than the coding region, and such modified genes are also included in the scope of the present invention. Thus, the present invention also encompasses polynucleotides and fragments of said genes having base sequences substantially the same as the original oncogene of SEQ ID NO: 9. Substantially identical polynucleotides refer to those DNAs encoding the same protein translation product as SEQ ID NO: 10 and having at least 80%, preferably at least 90%, most preferably at least 95% homology with SEQ ID NO: 9 do.

The protein expressed from the proto-oncogene of the present invention consists of 334 amino acids and has an amino acid sequence set forth in SEQ ID NO: 10 and is about 37 kDa in size. However, even in the amino acid sequence of the protein, substitution, addition or deletion of amino acids may be made within a range that does not affect the function of the protein, and only a part of the protein may be used depending on the purpose. Such modified amino acid sequences are also within the scope of the present invention. Accordingly, the present invention also encompasses polypeptides and fragments thereof having substantially the same amino acid sequence as the carcinogenic protein, wherein substantially identical polypeptides are at least 80%, preferably at least 90% and most preferably at least 95% sequences It means things with same sex.

4. TRG  6

Human transformation-related gene 6 (hereinafter referred to as TRG6), which is a protocogene of the present invention, has an entire sequence of 3,309 bp in length as set forth in SEQ ID NO: 13. In the nucleotide sequence of SEQ ID NO: 13, an open reading frame corresponding to nucleotides 233 to 481 sites (479-481: end codon) is the entire protein coding region, and the amino acid sequence derived therefrom is SEQ ID NO: : Shown in 14 and consists of 82 amino acids (hereinafter referred to as "TRG6 protein").

The nucleotide sequence of SEQ ID NO: 13 was registered in the NIH GenBank database of the National Institutes of Health as registration number AY191222 (published: April 8, 2005). Some similarities of genes and gene sequences to RP11-175D17 clones on chromosome 9 were confirmed. However, this study identified TRG6 proto-oncogenes, which are highly expressed in various human tumors, including uterine cancer, and very low in various normal tissues.

 However, due to the degeneracy of the codon or in consideration of the preferred codon in the organism to express the primary cancer gene, the primary cancer gene of the present invention does not change the amino acid sequence of the carcinogenic protein expressed from the coding region. In the coding region, various modifications may be made, and various modifications or modifications may be made within a range that does not affect the expression of genes in portions other than the coding region, and such modified genes are also included in the scope of the present invention. Accordingly, the present invention also encompasses polynucleotides having fragments of the above genes and bases having substantially the same nucleotide sequence as the above oncogene of SEQ ID NO: 13. Substantially identical polynucleotides refer to those DNAs encoding the same protein translation product as SEQ ID NO: 14, having at least 80%, preferably at least 90%, most preferably at least 95% homology with SEQ ID NO: 13 do.

The protein expressed from the primary oncogene of the present invention is composed of 82 amino acids and has an amino acid sequence set forth in SEQ ID NO: 14 and is about 9 kDa in size. However, even in the amino acid sequence of the protein, substitution, addition or deletion of amino acids may be made within a range that does not affect the function of the protein, and only a part of the protein may be used depending on the purpose. Such modified amino acid sequences are also within the scope of the present invention. Accordingly, the present invention also encompasses polypeptides and fragments thereof having substantially the same amino acid sequence as the carcinogenic protein, wherein substantially identical polypeptides are at least 80%, preferably at least 90% and most preferably at least 95% sequences It means things with same sex.

5. TRG  7

Human transformation-related gene 7 (hereinafter referred to as TRG7), which is a protocogene of the present invention, has a full base sequence of 1,334 bp in length as set forth in SEQ ID NO: 17.

In the nucleotide sequence of SEQ ID NO: 17, the open reading frame corresponding to the nucleotide number 42 to 1422 site (1420-1422: end codon) is the entire protein coding region, and the amino acid sequence derived therefrom is SEQ ID NO: It is shown at 18 and consists of 175 amino acids (hereinafter referred to as "TRG7 protein").

The base sequence of SEQ ID NO: 17 was registered in the NIH GenBank database of the National Institutes of Health as registration number AY191223 (published: April 8, 2005). Homo sapiens cDNA FLJ90076 fis, clone HEMBA1004444 and similar gene sequences were confirmed. However, this study identified TRG7 proto-oncogenes, which are highly expressed in various human tumors, including uterine cancer, and very low in various normal tissues.

However, due to the degeneracy of the codon or in consideration of the preferred codon in the organism to express the primary cancer gene, the primary cancer gene of the present invention does not change the amino acid sequence of the carcinogenic protein expressed from the coding region. In the coding region, various modifications may be made, and various modifications or modifications may be made within a range that does not affect the expression of genes in portions other than the coding region, and such modified genes are also included in the scope of the present invention. Thus, the present invention also encompasses polynucleotides having fragments of the above genes and bases having substantially the same nucleotide sequence as the above oncogene of SEQ ID NO: 17. Substantially identical polynucleotides refer to DNAs encoding protein translation products identical to SEQ ID NO: 18, having those having at least 80%, preferably at least 90%, most preferably at least 95% homology with SEQ ID NO: 17 do.

The protein expressed from the primary oncogene of the present invention consists of 175 amino acids and has an amino acid sequence set forth in SEQ ID NO: 18 and is about 20 kDa in size. However, even in the amino acid sequence of the protein, substitution, addition or deletion of amino acids may be made within a range that does not affect the function of the protein, and only a part of the protein may be used depending on the purpose. Such modified amino acid sequences are also within the scope of the present invention. Accordingly, the present invention also encompasses polypeptides and fragments thereof having substantially the same amino acid sequence as the carcinogenic protein, wherein substantially identical polypeptides comprise at least 80%, preferably at least 90% and most preferably at least 95% sequences Means those with homology.

6. TRG  9

Human transformation-related gene 9 (hereinafter, referred to as TRG9), which is a protocogene of the present invention, has a full base sequence of 1,582 bp in length as set forth in SEQ ID NO: 21.

In the nucleotide sequence of SEQ ID NO: 21, the open reading frame corresponding to the nucleotide number 17 to 1576 site (1574-1576: end codon) is the entire protein coding region and the amino acid sequence derived therefrom is SEQ ID NO: It is shown at 22 and consists of 519 amino acids (hereinafter referred to as "TRG9 protein").

The base sequence of SEQ ID NO: 21 is registered in the NIH GenBank database of the National Institutes of Health as No. AY272044 (published: March 31, 2005). Homo sapiens sorting nexin 2 (SNX2) and similar gene sequences were identified. However, this research has led to the discovery of the TRG9 proto-oncogene, which is highly expressed in various human tumors, including uterine cancer, and very low in several normal tissues.

However, due to the degeneracy of the codon or in consideration of the preferred codon in the organism to express the primary cancer gene, the primary cancer gene of the present invention does not change the amino acid sequence of the carcinogenic protein expressed from the coding region. In the coding region, various modifications may be made, and various modifications or modifications may be made within a range that does not affect the expression of genes in portions other than the coding region, and such modified genes are also included in the scope of the present invention. Thus, the present invention also encompasses polynucleotides having fragments of the above genes and bases having substantially the same nucleotide sequence as the above oncogene of SEQ ID NO: 21. Substantially identical polynucleotides refer to those DNAs encoding the same protein translation product as SEQ ID NO: 22, having at least 80%, preferably at least 90%, most preferably at least 95% homology with SEQ ID NO: 21 do.

The protein expressed from the proto-oncogene of the present invention consists of 519 amino acids, has an amino acid sequence set forth in SEQ ID NO: 22, and is about 58 kDa in size. However, even in the amino acid sequence of the protein, substitution, addition or deletion of amino acids may be made within a range that does not affect the function of the protein, and only a part of the protein may be used depending on the purpose. Such modified amino acid sequences are also within the scope of the present invention. Accordingly, the present invention also encompasses polypeptides and fragments thereof having substantially the same amino acid sequence as the carcinogenic protein, wherein substantially identical polypeptides are at least 80%, preferably at least 90% and most preferably at least 95% sequences It means things with same sex.

7. TRG  10

Human transformation-related gene 10 (hereinafter, referred to as TRG10), which is a protocogene of the present invention, has an entire sequence of 3,979 bp in length as set forth in SEQ ID NO: 25.

In the nucleotide sequence of SEQ ID NO: 25, the open reading frame corresponding to the nucleotide number 1100-1270 site (1268-1270: end codon) is the entire protein coding region and the amino acid sequence derived therefrom is SEQ ID NO: It is shown in 26 and consists of 56 amino acids (hereinafter referred to as "TRG10 protein").

The base sequence of SEQ ID NO: 25 was registered as Registration Number AY277593 in the NIH GenBank Database of the National Institutes of Health (published: March 31, 2005). Homo sapiens bone morphogenetic protein receptor, type II (serine / threonine kinase) (BMPR2), transcriptional variant 2 gene and similar gene sequences were identified. However, this study has led to the discovery of the TRG10 proto-oncogene, which is highly expressed in many human tumors, including uterine cancer, and low in many normal tissues.

However, due to the degeneracy of the codon or in consideration of the preferred codon in the organism to express the primary cancer gene, the primary cancer gene of the present invention does not change the amino acid sequence of the carcinogenic protein expressed from the coding region. In the coding region, various modifications may be made, and various modifications or modifications may be made within a range that does not affect the expression of genes in portions other than the coding region, and such modified genes are also included in the scope of the present invention. Thus, the present invention also encompasses polynucleotides having fragments of the above genes and bases having substantially the same nucleotide sequence as the above oncogene of SEQ ID NO: 25. Substantially identical polynucleotides are DNAs encoding a protein translation product identical to SEQ ID NO: 26, having those having sequence homology at least 80%, preferably at least 90%, most preferably at least 95% with SEQ ID NO: 25 it means.

The protein expressed from the proto-oncogene of the present invention consists of 56 amino acids and has an amino acid sequence set forth in SEQ ID NO: 26 and is about 6 kDa in size. However, even in the amino acid sequence of the protein, substitution, addition or deletion of amino acids may be made within a range that does not affect the function of the protein, and only a part of the protein may be used depending on the purpose. Such modified amino acid sequences are also within the scope of the present invention. Accordingly, the present invention also encompasses polypeptides and fragments thereof having substantially the same amino acid sequence as the carcinogenic protein, wherein substantially identical polypeptides are at least 80%, preferably at least 90% and most preferably at least 95% sequences It means things with same sex.

8. TRG  11

Human transformation-related gene 11 (hereinafter, referred to as TRG11), which is a protocogene of the present invention, has an entire base sequence of 235 bp in length as set forth in SEQ ID NO: 29.

In the nucleotide sequence of SEQ ID NO: 29, the open reading frame corresponding to the nucleotide numbers 26 to 214 sites (227-229: end codon) is the entire protein coding region and the amino acid sequence derived therefrom is SEQ ID NO: It is shown at 30 and consists of 62 amino acids (hereinafter referred to as "TRG11 protein").

The base sequence of SEQ ID NO: 29 was registered as registered number AY277594 in the NIH GenBank database bay of the National Institutes of Health (published: March 31, 2005). Homo sapiens IMAGE: 5258564 Gene sequences and similar gene sequences were confirmed. However, this study identified TRG11 proto-oncogenes, which are highly expressed in various human tumors, including uterine cancer, but very low in various normal tissues.

However, due to the degeneracy of the codon or in consideration of the preferred codon in the organism to express the primary cancer gene, the primary cancer gene of the present invention does not change the amino acid sequence of the carcinogenic protein expressed from the coding region. In the coding region, various modifications may be made, and various modifications or modifications may be made within a range that does not affect the expression of genes in portions other than the coding region, and such modified genes are also included in the scope of the present invention. Accordingly, the present invention also encompasses polynucleotides having fragments of the above genes and bases having substantially the same nucleotide sequence as the above oncogene of SEQ ID NO: 29. Substantially identical polynucleotides refer to those DNAs encoding a protein translation product identical to SEQ ID NO: 30, having at least 80%, preferably at least 90%, most preferably at least 95% homology with SEQ ID NO: 29 do.

The protein expressed from the proto-oncogene of the present invention consists of 62 amino acids, has an amino acid sequence set forth in SEQ ID NO: 30, and is about 7 kDa in size. However, even in the amino acid sequence of the protein, substitution, addition or deletion of amino acids may be made within a range that does not affect the function of the protein, and only a part of the protein may be used depending on the purpose. Such modified amino acid sequences are also within the scope of the present invention. Accordingly, the present invention also encompasses polypeptides and fragments thereof having substantially the same amino acid sequence as the carcinogenic protein, wherein substantially identical polypeptides are at least 80%, preferably at least 90% and most preferably at least 95% sequences It means things with same sex.

9. TRG  12

Human transformation-related gene 12 (hereinafter referred to as TRG12), which is a protocogene of the present invention, has an entire sequence of 510 bp in length as set forth in SEQ ID NO: 33.

In the nucleotide sequence of SEQ ID NO: 33, the open reading frame corresponding to nucleotide numbers 80 to 475 (473-475: end codon) is the entire protein coding region, and the amino acid sequence derived therefrom is SEQ ID NO: It is shown at 34 and consists of 131 amino acids (hereinafter referred to as "TRG12 protein").

The base sequence of SEQ ID NO: 33 is registered as Registration No. AY277595 in the NIH GenBank Database of the National Institutes of Health (published: March 31, 2005). Homo sapiens DNA59497 MY047 (UNQ577) It was confirmed that the gene sequence and the like are similar. However, this study identified TRG12 proto-oncogenes, which are highly expressed in various human tumors, including uterine cancer, and very low in various normal tissues.

However, due to the degeneracy of the codon or in consideration of the preferred codon in the organism to express the primary cancer gene, the primary cancer gene of the present invention does not change the amino acid sequence of the carcinogenic protein expressed from the coding region. In the coding region, various modifications may be made, and various modifications or modifications may be made within a range that does not affect the expression of genes in portions other than the coding region, and such modified genes are also included in the scope of the present invention. Accordingly, the present invention also encompasses polynucleotides having fragments of the above genes and bases having substantially the same nucleotide sequence as the above oncogene of SEQ ID NO: 33. Substantially identical polynucleotides refer to those DNAs encoding the same protein translation product as SEQ ID NO: 34, having at least 80%, preferably at least 90%, most preferably at least 95% homology with SEQ ID NO: 33 do.

The protein expressed from the proto-oncogene of the present invention consists of 131 amino acids and has an amino acid sequence set forth in SEQ ID NO: 34 and is about 14 kDa in size. However, even in the amino acid sequence of the protein, substitution, addition or deletion of amino acids may be made within a range that does not affect the function of the protein, and only a part of the protein may be used depending on the purpose. Such modified amino acid sequences are also within the scope of the present invention. Accordingly, the present invention also encompasses polypeptides and fragments thereof having substantially the same amino acid sequence as the carcinogenic protein, wherein substantially identical polypeptides are at least 80%, preferably at least 90% and most preferably at least 95% sequences It means things with same sex.

10. TRG  13

Human transformation-related gene 13 (hereinafter referred to as TRG13), which is a protocogene of the present invention, has a full base sequence of 1,301 bp in length as set forth in SEQ ID NO: 37.

In the nucleotide sequence of SEQ ID NO: 37, the open reading frame corresponding to nucleotide number 18 to 1193 region (1191-1193: end codon) is the entire protein coding region, and the amino acid sequence derived therefrom is SEQ ID NO: It is shown at 38 and consists of 391 amino acids (hereinafter referred to as "TRG13 protein").

The base sequence of SEQ ID NO: 37 was registered as Registration No. AY277596 with the NIH GenBank database of the National Institutes of Health (published: March 31, 2005). Homo sapiens MGC11349 gene and the Homo sapiens MGC11349 gene, BC012729 gene, were identified to be similar. These genes are not yet known. However, this study identified TRG13 proto-oncogenes, which are highly expressed in various human tumors, including uterine cancer, and very low in various normal tissues.

However, due to the degeneracy of the codon or in consideration of the preferred codon in the organism to express the primary cancer gene, the primary cancer gene of the present invention does not change the amino acid sequence of the carcinogenic protein expressed from the coding region. In the coding region, various modifications may be made, and various modifications or modifications may be made within a range that does not affect the expression of genes in portions other than the coding region, and such modified genes are also included in the scope of the present invention. Thus, the present invention also encompasses polynucleotides having fragments of the above genes and bases having substantially the same nucleotide sequences as those of the original oncogene of SEQ ID NO: 37. Substantially identical polynucleotides refer to those DNAs encoding the same protein translation product as SEQ ID NO: 38 and having at least 80%, preferably at least 90% and most preferably at least 95% sequence homology with SEQ ID NO: 37 do.

The protein expressed from the proto-oncogene of the present invention consists of 391 amino acids and has an amino acid sequence set forth in SEQ ID NO: 38 and is about 40 kDa in size. However, even in the amino acid sequence of the protein, substitution, addition or deletion of amino acids may be made within a range that does not affect the function of the protein, and only a part of the protein may be used depending on the purpose. Such modified amino acid sequences are also within the scope of the present invention. Accordingly, the present invention also encompasses polypeptides and fragments thereof having substantially the same amino acid sequence as the carcinogenic protein, wherein substantially identical polypeptides are at least 80%, preferably at least 90% and most preferably at least 95% sequences It means things with same sex.

11. TRG  14

Human transformation-related gene 14 (hereinafter referred to as TRG14), which is a protocogene of the present invention, has an entire base sequence of 1,206 bp in length as set forth in SEQ ID NO: 41.

In the nucleotide sequence of SEQ ID NO: 41, the open reading frame corresponding to the nucleotide number 18 to 1202 region (1200-1202: end codon) is the entire protein coding region and the amino acid sequence derived therefrom is SEQ ID NO: It is shown at 42 and consists of 394 amino acids (hereinafter referred to as "TRG14 protein").

The base sequence of SEQ ID No. 41 is registered in the NIH GenBank Database of the National Institutes of Health as No. AY277597 (published: March 31, 2005). As a result of sequencing analysis, NM_006096 Homo sapiens N-myc downstream regulated gene 1 (NDRG1) gene and similar gene sequence was confirmed. However, this study identified TRG14 proto-oncogenes, which are highly expressed in various human tumors, including uterine cancer, but very low in several normal tissues.

However, due to the degeneracy of the codon or in consideration of the preferred codon in the organism to express the primary cancer gene, the primary cancer gene of the present invention does not change the amino acid sequence of the carcinogenic protein expressed from the coding region. In the coding region, various modifications may be made, and various modifications or modifications may be made within a range that does not affect the expression of genes in portions other than the coding region, and such modified genes are also included in the scope of the present invention. Thus, the present invention also encompasses polynucleotides having fragments of the above genes and bases having substantially the same nucleotide sequence as the above oncogene of SEQ ID NO: 41. Substantially identical polynucleotides are DNAs encoding a protein translation product identical to SEQ ID NO: 42 and those having sequence homology with at least 80%, preferably at least 90% and most preferably at least 95% with SEQ ID NO: 41. it means.

The protein expressed from the proto-oncogene of the present invention consists of 394 amino acids and has an amino acid sequence set forth in SEQ ID NO: 42 and is about 43 kDa in size. However, even in the amino acid sequence of the protein, substitution, addition or deletion of amino acids may be made within a range that does not affect the function of the protein, and only a part of the protein may be used depending on the purpose. Such modified amino acid sequences are also within the scope of the present invention. Accordingly, the present invention also encompasses polypeptides and fragments thereof having substantially the same amino acid sequence as the carcinogenic protein, wherein substantially identical polypeptides are at least 80%, preferably at least 90% and most preferably at least 95% sequences It means things with same sex.

12. TRG  15

Human transformation-related gene 15 (hereinafter referred to as TRG15), which is a protocogene of the present invention, has an entire base sequence of 1,104 bp in length as set forth in SEQ ID NO: 45.

In the nucleotide sequence of SEQ ID 45, the open reading frame corresponding to the nucleotide numbers 1 to 1104 sites (1102-1104: end codon) is the entire protein coding region and the amino acid sequence derived therefrom is SEQ ID NO: It is shown at 46 and consists of 367 amino acids (hereinafter referred to as "TRG15 protein").

The base sequence of SEQ ID NO: 45 was registered as Registration No. AY277598 in the NIH GenBank Database of the National Institutes of Health (published: March 31, 2005). Homo sapiens FLJ20758 protein gene NM_017952 was found to be similar to the gene sequence. The FLJ20758 protein gene is not yet known. However, this study identified TRG15 proto-oncogenes, which are highly expressed in various human tumors, including uterine cancer, and very low in various normal tissues.

However, due to the degeneracy of the codon or in consideration of the preferred codon in the organism to express the primary cancer gene, the primary cancer gene of the present invention does not change the amino acid sequence of the carcinogenic protein expressed from the coding region. In the coding region, various modifications may be made, and various modifications or modifications may be made within a range that does not affect the expression of genes in portions other than the coding region, and such modified genes are also included in the scope of the present invention. Accordingly, the present invention also encompasses polynucleotides having fragments of the above genes and bases having substantially the same nucleotide sequence as the above oncogene of SEQ ID NO: 45. Substantially identical polynucleotides refer to those DNAs encoding the same protein translation product as SEQ ID NO: 46, having at least 80%, preferably at least 90%, most preferably at least 95% homology with SEQ ID NO: 45 do.

The protein expressed from the proto-oncogene of the present invention consists of 367 amino acids and has an amino acid sequence set forth in SEQ ID NO: 46 and is about 42 kDa in size. However, even in the amino acid sequence of the protein, substitution, addition or deletion of amino acids may be made within a range that does not affect the function of the protein, and only a part of the protein may be used depending on the purpose. Such modified amino acid sequences are also within the scope of the present invention. Accordingly, the present invention also encompasses polypeptides and fragments thereof having substantially the same amino acid sequence as the carcinogenic protein, wherein substantially identical polypeptides are at least 80%, preferably at least 90% and most preferably at least 95% sequences It means things with same sex.

13. TRG  16

Human transformation-related gene 16 (hereinafter referred to as TRG16), which is a protocogene of the present invention, has a full base sequence of 1,064 bp in length represented by SEQ ID NO: 49.

In the nucleotide sequence of SEQ ID NO: 49, the open reading frame corresponding to the nucleotide number 92 to 1064 region (1062-1064: end codon) is the entire protein coding region, and the amino acid sequence derived therefrom is SEQ ID NO: It is shown at 50 and consists of 324 amino acids (hereinafter referred to as "TRG16 protein").

The base sequence of SEQ ID NO: 49 was registered in the NIH GenBank Database of the National Institutes of Health as Registration No. AY277601 (published: March 31, 2005). Homo sapiens pp9320 mRNA gene and gene sequence were confirmed to be similar. The pp9320 gene is not yet known. However, this study identified TRG16 proto-oncogenes, which are highly expressed in various human tumors, including uterine cancer, and very low in various normal tissues.

However, due to the degeneracy of the codon or in consideration of the preferred codon in the organism to express the primary cancer gene, the primary cancer gene of the present invention does not change the amino acid sequence of the carcinogenic protein expressed from the coding region. In the coding region, various modifications may be made, and various modifications or modifications may be made within a range that does not affect the expression of genes in portions other than the coding region, and such modified genes are also included in the scope of the present invention. Accordingly, the present invention also encompasses polynucleotides having fragments of the above genes and bases having substantially the same nucleotide sequence as the above-described oncogene of SEQ ID NO: 49. Substantially identical polynucleotide means DNAs encoding a protein translation product identical to SEQ ID NO: 50, having those having at least 80%, preferably at least 90%, most preferably at least 95% homology with SEQ ID NO: 49 do.

The protein expressed from the proto-oncogene of the present invention consists of 324 amino acids and has an amino acid sequence set forth in SEQ ID NO: 50 and is about 36 kDa in size. However, even in the amino acid sequence of the protein, substitution, addition or deletion of amino acids may be made within a range that does not affect the function of the protein, and only a part of the protein may be used depending on the purpose. Such modified amino acid sequences are also within the scope of the present invention. Accordingly, the present invention also encompasses polypeptides and fragments thereof having substantially the same amino acid sequence as the carcinogenic protein, wherein substantially identical polypeptides are at least 80%, preferably at least 90% and most preferably at least 95% sequences It means things with same sex.

14. TRG  17

Human transformation-related gene 17 (hereinafter referred to as TRG17), which is a protocogene of the present invention, has an entire base sequence of 432 bp in length as set forth in SEQ ID NO: 53.

In the nucleotide sequence of SEQ ID NO: 53, the open reading frame corresponding to nucleotide numbers 1 to 408 (406-408: end codon) is the entire protein coding region, and the amino acid sequence derived therefrom is SEQ ID NO: It is shown at 54 and consists of 135 amino acids (hereinafter referred to as "TRG17 protein"). The base sequence of SEQ ID NO: 53 is registered as Registration No. AY277599 in the NIH GenBank Database of the National Institutes of Health (published: March 31, 2005). Homo sapiens MGC5309 (MGC5309) gene and gene sequence BC003353 Homo sapiens MGC5309 gene was confirmed to be similar. However, this study identified TRG17 proto-oncogenes, which are highly expressed in various human tumors, including uterine cancer, and very low in various normal tissues.

However, due to the degeneracy of the codon or in consideration of the preferred codon in the organism to express the primary cancer gene, the primary cancer gene of the present invention does not change the amino acid sequence of the carcinogenic protein expressed from the coding region. In the coding region, various modifications may be made, and various modifications or modifications may be made within a range that does not affect the expression of genes in portions other than the coding region, and such modified genes are also included in the scope of the present invention. Accordingly, the present invention also encompasses polynucleotides having fragments of the same genes and bases having substantially the same base sequence as the original cancer gene of SEQ ID NO: 53. Substantially identical polynucleotides are DNAs encoding a protein translation product identical to SEQ ID NO: 54, meaning those having sequence homology of at least 80%, preferably at least 90%, most preferably at least 95% with SEQ ID NO: 53 do.

The protein expressed from the primary oncogene of the present invention consists of 135 amino acids and has an amino acid sequence as set out in SEQ ID NO: 54 and is about 16 kDa in size. However, even in the amino acid sequence of the protein, substitution, addition or deletion of amino acids may be made within a range that does not affect the function of the protein, and only a part of the protein may be used depending on the purpose. Such modified amino acid sequences are also within the scope of the present invention. Accordingly, the present invention also encompasses polypeptides and fragments thereof having substantially the same amino acid sequence as the carcinogenic protein, wherein substantially identical polypeptides are at least 80%, preferably at least 90% and most preferably at least 95% sequences It means things with same sex.

15. TRG  18

Human transformation-related gene 18 (hereinafter, referred to as TRG18), which is a protocogene of the present invention, has a full base sequence of 1,141 bp in length as set forth in SEQ ID NO: 57.

In the nucleotide sequence of SEQ ID NO: 57, the open reading frame corresponding to the nucleotide number 20 to 1141 site (1139-1141: end codon) is the entire protein coding region and the amino acid sequence derived therefrom is SEQ ID NO: It is shown at 58 and consists of 373 amino acids (hereinafter referred to as "TRG18 protein"). The base sequence of SEQ ID NO: 57 was registered as Registration Number AY277600 in the NIH GenBank Database of the National Institutes of Health (published: March 31, 2005). Homo sapiens PEX3 protein homologous gene sequences and similar genes were confirmed. However, this study identified TRG18 proto-oncogenes, which are highly expressed in various human tumors, including uterine cancer, and very low in various normal tissues.

However, due to the degeneracy of the codon or in consideration of the preferred codon in the organism to express the primary cancer gene, the primary cancer gene of the present invention does not change the amino acid sequence of the carcinogenic protein expressed from the coding region. In the coding region, various modifications may be made, and various modifications or modifications may be made within a range that does not affect the expression of genes in portions other than the coding region, and such modified genes are also included in the scope of the present invention. Accordingly, the present invention also encompasses polynucleotides having fragments of the above genes and bases having substantially the same nucleotide sequence as the above oncogene of SEQ ID NO: 57. Substantially identical polynucleotides are DNAs encoding a protein translation product identical to SEQ ID NO: 58, meaning those having sequence homology of at least 80%, preferably at least 90%, most preferably at least 95% with SEQ ID NO: 57 do.

The protein expressed from the proto-oncogene of the present invention consists of 373 amino acids and has an amino acid sequence set forth in SEQ ID NO: 58 and is about 42 kDa in size. However, even in the amino acid sequence of the protein, substitution, addition or deletion of amino acids may be made within a range that does not affect the function of the protein, and only a part of the protein may be used depending on the purpose. Such modified amino acid sequences are also within the scope of the present invention. Accordingly, the present invention also encompasses polypeptides and fragments thereof having substantially the same amino acid sequence as the carcinogenic protein, wherein substantially identical polypeptides are at least 80%, preferably at least 90% and most preferably at least 95% sequences It means things with same sex.

16. TRG  20

Human transformation-related gene 20 (hereinafter, referred to as TRG20), which is a protocogene of the present invention, has an entire sequence of 449 bp in length as set forth in SEQ ID NO: 61.

In the nucleotide sequence of SEQ ID NO: 61, the open reading frame corresponding to nucleotide number 42 to 449 site (447-449: end codon) is the entire protein coding region, and the amino acid sequence derived therefrom is SEQ ID NO: It is shown at 62 and consists of 135 amino acids (hereinafter referred to as "TRG20 protein"). The base sequence of SEQ ID NO: 61 was registered as Registration Number AY453397 in the NIH GenBank Database of the National Institutes of Health (published: March 31, 2005). Homo sapiens hypothetical protein MGC5309 (MGC5309) gene and BC003353 Homo sapiens hypothetical protein MGC5309 gene was confirmed to be similar to the gene sequence. However, this study identified TRG20 proto-oncogenes, which are highly expressed in various human tumors, including uterine cancer, but very low in various normal tissues.

However, due to the degeneracy of the codon or in consideration of the preferred codon in the organism to express the primary cancer gene, the primary cancer gene of the present invention does not change the amino acid sequence of the carcinogenic protein expressed from the coding region. In the coding region, various modifications may be made, and various modifications or modifications may be made within a range that does not affect the expression of genes in portions other than the coding region, and such modified genes are also included in the scope of the present invention. Accordingly, the present invention also encompasses polynucleotides having fragments of the above genes and bases having substantially the same nucleotide sequence as the above oncogene of SEQ ID NO: 61. Substantially identical polynucleotides refer to those DNAs encoding the same protein translation product as SEQ ID NO: 62 and having at least 80%, preferably at least 90% and most preferably at least 95% homology with SEQ ID NO: 61 do.

The protein expressed from the proto-oncogene of the present invention consists of 135 amino acids, has an amino acid sequence set forth in SEQ ID NO: 62, and is about 16 kDa in size. However, even in the amino acid sequence of the protein, substitution, addition or deletion of amino acids may be made within a range that does not affect the function of the protein, and only a part of the protein may be used depending on the purpose. Such modified amino acid sequences are also within the scope of the present invention. Accordingly, the present invention also encompasses polypeptides and fragments thereof having substantially the same amino acid sequence as the carcinogenic protein, wherein substantially identical polypeptides are at least 80%, preferably at least 90% and most preferably at least 95% sequences It means things with same sex.

The original cancer genes and proteins of the present invention may be isolated from human cancer tissues or synthesized according to known methods of DNA or peptide synthesis. In addition, such a gene can be inserted into a microorganism expression vector known in the art to prepare an expression vector, and then introduced into an appropriate host cell, for example, E. coli or yeast cell. Using the transformed host as described above, the DNA of the gene of the present invention can be opposed to each other or a large amount of protein can be produced.

When constructing a vector, it is possible to appropriately select and combine expression control regulatory sequences such as promoters, terminators, self-replicating sequences and secretion signals, depending on the type of host cell to produce the above-described source cancer gene or protein.

The gene of the present invention is considered to be a powerful oncogenic gene that induces uterine cancer because of overexpression in cervical cancer tissues and uterine cervical cancer cell lines in normal cervical tissues, whereas in normal cervical tissue analysis methods such as Northern blot. In addition to epithelial tissues such as cervical cancer, the primary cancer genes of the present invention are highly expressed in many other cancers such as leukemia and colorectal cancer. Thus, the original cancer gene of the present invention is judged to be a carcinogenic gene common to various cancer occurrences, and can be effectively used for diagnosis of various cancers, production of transformed animals, anti-sense gene therapy, and the like.

For example, the method for diagnosing cancer using the source cancer gene may be performed by detecting all kinds or parts of the source cancer gene by using a probe to hybridize with the nucleic acid separated from the body fluid of the subject, and then detecting them by various methods known in the art. Includes a process of judging whether the gene contains the primary cancer gene of the present invention. Labeling the probe with a radioisotope or enzyme can facilitate identification of the presence of the gene. Accordingly, the present invention also provides a kit for diagnosing cancer comprising all or a portion of the source cancer gene.

Transgenic animals can be produced by introducing the original cancer gene of the present invention into mammals, for example, rodents such as rats, and the introduction of this gene at least at the stage of fertilization before the eight-cell stage is desirable. The transgenic animals thus produced can be usefully used for the search for anti-cancer substances such as carcinogens or antioxidants.

The present invention also provides anti-sense genes that are effective in gene therapy. As used herein, an “anti-sense gene” is a polynucleotide having a sequence complementary to a portion or entire portion of an mRNA that is transcribed from the primary cancer gene or a portion thereof, and binds to the protein binding site of the mRNA. By introducing DNA sequences with sequences capable of destroying open reading frames (ORFs) into the patient's body, cancer can be prevented or treated due to the expression of the original cancer gene. The present invention also encompasses a method for treating or preventing cancer or cancer metastasis by administering to a patient a therapeutically effective amount of an antisense gene of the invention within its scope.

In the antisense gene therapy of the present invention, the antisense gene of the present invention is administered to a patient in a conventional manner to prevent the expression of the proto-oncogene. See, eg, J. S. Kim et al., J. Controlled 　 Antisense oligodeoxynucleotides (ODN) are mixed with poly-L-lysine derivatives by electrostatic attraction according to the method of Release 53: 175-182, 1998) and the mixture is administered intravenously to the patient.

The scope of the present invention also includes anticancer compositions comprising the antisense gene of the present invention as an active ingredient together with pharmaceutically acceptable carriers, excipients or, optionally, other additives. It is desirable to formulate the pharmaceutical compositions of the present invention into injections.

The amount of antisense gene that is actually administered is determined in consideration of a variety of related factors such as the symptom being treated, the route of administration, the age and weight of the patient, and the severity of the symptoms.

Proteins derived from the source cancer gene of the present invention can be usefully used as diagnostic tools to produce antibodies. Antibodies of the present invention can be prepared as monoclonal antibodies or polyclonal antibodies according to conventional methods known in the art, using proteins or their fragments having the above amino acid sequences expressed from the original cancer gene of the present invention. Enzyme immunoassays (ELISA), radioimmunoassay (RIA), sandwich assays (sandwich assays) on polyacrylamide gels, and the like using immunosorbents known in the art using such antibodies According to the method, cancer can be diagnosed by confirming whether the protein is expressed in the body fluid sample of the subject.

In addition, it is possible to establish cancer cell lines capable of continuously proliferating by using the original cancer gene of the present invention.These cell lines are examples of tumor tissues that are formed on the nucleus of the mouse by using the fibroblasts transduced with the primary cancer gene. It can be manufactured from. These cancer cell lines can be usefully used for the detection of anticancer drugs.

Hereinafter, the present invention will be described in detail with reference to Examples.

However, the following examples are merely illustrative of the present invention, and the contents of the present invention are not limited to the following examples.

Example 1 Isolation of Tumor Cells and Isolation of Total RNA

(Step 1) 양 Culture of tumor cells

For differential development of mRNA, primary cervical tumor tissues and metastases during surgery from uterine myomas undergoing hysterectomy and from uterine cervical tissue and from uterine cancer patients previously untreated with chemotherapy and radiation Lymph node tumors were taken. Differential development includes CUMC-6 (Kim, J. W. et. 　 al . , Gynecol . Oncol . 62: 230-240, 1996).

The cells obtained from the tissues obtained and the CUMC-6 cell line were obtained by `` MBM '' glutamine, 100 IU / mL penicillin, 100 μg / mL streptomycin and 10% fetal bovine serum (Gibco, USA). Proliferation was carried out in 752/1 culture medium (Gibco). The cultured cells used in the experiments were cells that showed 95% or more viability by trypan blue staining as exponential growth cells (Freshney, "Culture of Animal Cells: A Manual of Basic Technique" 2nd). Ed., AR Liss, New York, 1987).

(Step 2) Isolation of RNA Differentiation of mRNA Development method

Total RNA was extracted from normal uterine cervical tissue, primary uterine cervical tumor tissue, metastatic lymph node tumor tissue and CUMC-6 cells obtained in stage 1 using the commercially available RNeasy Total RNA Kit (Qiagen Inc., Germany). . DNA contaminants were removed from the RNA using a Message clean kit (GenHunter Corp., Brookline, MA).

Example 2 Differential Development Reverse Transcription Polymerase Chain Reaction (differential display reverse-transcription-PCR)

Differential development reverse transcription was performed by slightly modifying the reverse transcription-polymerase chain reaction (RT-PCR) described by Liang and Paddy.

2-1: TRG  3

First, H-T11A fixed primer (5'-AAGCTTTTTTTTTTTA-3 ', RNAimage kit having a nucleotide sequence shown as SEQ ID NO: 3 as an oligo-dT primer immobilized on 0.2 μg of total RNA obtained in step 1 of Example 1) , Genhunter, Cor., MA, USA) was used to perform reverse transcription.

Then, in the presence of 0.5 mM [α-35S] dATP (1200 Ci / mmole), the same fixed primers and random 5 ′ 11 primers (RNAimage primer sets 1-5) described as SEQ ID NO: 4 in H-APs 1-40 PCR was performed using H-AP20 primer (5'-AAGCTTGTTGTGC-3 ') having a nucleotide sequence. The PCR reaction conditions were 40 seconds at 95 ° C., 40 minutes at 40 ° C. for 2 minutes, and 40 seconds at 72 ° C. for 5 minutes at 72 ° C. for final extension.

After PCR-amplified fragments were dissolved in 6% polyacrylamide sequencing gels, radiographs were used to determine the position of the bands showing different degrees of differential expression.

A 258 base pair (bp) sized band, H20-121 cDNA (SEQ ID NO: 1342-1599 bp) was cut out from the dried gel. After elution of H20-121 cDNA by heating for 15 minutes, PCR was performed under the same primers and under the same conditions except that [α-35S] labeled dATP (1200 Ci / mmole) and 20 μM dNTP were not used. By reamplification.

2-2: TRG  4

H-T11G® immobilized primers (SEQ ID NO: 7) (5'-AAGCTTTTTTTTTTTG-3 ', RNAimage kit, Genhunter, Cor., MA, USA) and H-AP9 primers (SEQ ID NO: 8) (5'-AAGCTTCATTCCG-3') PCR was carried out in the same manner as in the above 2-1, except that was used.

A 393 base pair (bp) sized band, H93-811 cDNA (SEQ ID NO: 2086-2478 bp) was cut out from the dried gel. After eluting H93-811 cDNA by heating for 15 minutes, PCR was carried out under the same primers and under the same conditions except that [α- ³⁵ S] labeled dATP (1200 Ci / mmole) and 20 μM dNTP were not used. And reamplified.

2-3: TRG  5

H-T11C fixed primer (5'-AAGCTTTTTTTTTTTC-3 ', RNAimage kit, Genhunter, Cor., MA, USA) having base sequence set forth in SEQ ID NO: 11 and H having base sequence set forth in SEQ ID NO: 12 PCR was performed by the same procedure as in 2-1, except that -AP11 primer (5'-AAGCTTCGGGTAA-3 ') was used.

A band of 292 base pairs (bp) size, H117-321 cDNA (SEQ ID NO: 933-1224 bp) was cut out from the dried gel. After eluting H117-321 cDNA by heating for 15 minutes, PCR was performed under the same primers and under the same conditions except that [α-35S] labeled dATP (1200 Ci / mmole) and 20 μM dNTP were not used. By reamplification.

2-4: TRG  6

H-T11A fixed primer having the nucleotide sequence set forth in SEQ ID NO: 15 (5'- AAGCTTTTTTTTTTTA-3 ', RNAimage kit, Genhunter, Cor., MA, USA) and H having the nucleotide sequence set forth in SEQ ID NO: 16 PCR was carried out in the same manner as in 2-1, except that -AP38 primer (5'-AAGCTTCCAGTGC-3 ') was used.

A 311 base pair (bp) sized band, H38-211 cDNA (2823-3133 bp of SEQ ID NO: 13) was cut out from the dried gel. After eluting H38-211 cDNA by heating for 15 minutes, PCR was carried out under the same primers and under the same conditions except that [α-35S] labeled dATP (1200 Ci / mmole) and 20 μM dNTP were not used. By reamplification.

2-5: TRG  7

H-T11G fixed primer having the nucleotide sequence set forth in SEQ ID NO: 19 (5'-AAGCTTTTTTTTTTTG-3 ', RNAimage kit, Genhunter, Cor., MA, USA) and H having the nucleotide sequence set forth in SEQ ID NO: 20 PCR was carried out in the same manner as in 2-1, except that -AP38 primer (5'-AAGCTTCCAGTGC-3 ') was used.

A 292 base pair (bp) sized band, H38-621 cDNA (SEQ ID NO: 1404-1695 bp) was cut out from the dried gel. After eluting H38-621 cDNA by heating for 15 minutes, PCR was performed under the same primers and under the same conditions except that [α-35S] labeled dATP (1200 Ci / mmole) and 20 μM dNTP were not used. By reamplification.

2-6: TRG  9

H-T11A fixed primer having the nucleotide sequence set forth in SEQ ID NO: 23 (5'-AAGCTTTTTTTTTTTA-3 ', RNAimage kit, Genhunter, Cor., MA, USA) and H having the nucleotide sequence set forth in SEQ ID NO: 24 PCR was carried out in the same manner as in 2-1, except that -AP9 primer (5'-AAGCTTCATTCCG-3 ') was used.

A 275 base pair (bp) sized band, H96 cDNA (SEQ ID NO: 1225-1499 bp) was cut out from the dried gel. After heating for 15 minutes to elute H96 cDNA, PCR was performed under the same primers and the same conditions as above except that [α-35S] labeled dATP (1200 Ci / mmole) and 20 μM dNTP were not used. Amplified.

2-7; TRG10

H-T11A fixed primer having the nucleotide sequence set forth in SEQ ID NO: 27 (5'-AAGCTTTTTTTTTTTA-3 ', RNAimage kit, Genhunter, Cor., MA, USA) and H having the nucleotide sequence set forth in SEQ ID NO: 28 PCR was performed by the same procedure as in 2-1 except that the AP9 primer (5'-AAGCTTCATTCCG-3 ') was used.

A 352 base pair (bp) sized band, H94 cDNA (SEQ ID NO: 3528-3879 bp) was cut out from the dried gel. After eluting H94 cDNA by heating for 15 minutes, PCR was performed under the same primers and the same conditions as above except that [α-35S] labeled dATP (1200 Ci / mmole) and 20 μM dNTP were not used. Amplified.

2-8; TRG11

H-T11C fixed primer having the nucleotide sequence set forth in SEQ ID NO: 31 (5'-AAGCTTTTTTTTTTTC-3 ', RNAimage kit, Genhunter, Cor., MA, USA) and H having the nucleotide sequence set forth in SEQ ID NO: 32 PCR was performed by the same procedure as in 2-1, except that -AP4 primer (5'-AAGCTTCTCAACG-3 ') was used.

A 147 base pair (bp) sized band, H42 cDNA (83-229 bp of SEQ ID NO: 29) was cut out from the dried gel. After eluting H42 cDNA by heating for 15 minutes, PCR was performed under the same primers and the same conditions as above except that [α-35S] labeled dATP (1200 Ci / mmole) and 20 μM dNTP were not used. Amplified.

2-9: TRG  12

H-T11G fixed primer having the nucleotide sequence set forth in SEQ ID NO: 35 (5'-AAGCTTTTTTTTTTTG-3 ', RNAimage kit, Genhunter, Cor., MA, USA) and H having the nucleotide sequence set forth in SEQ ID NO: 36 PCR was performed by the same procedure as in 2-1 except that the AP10 primer (5'-AAGCTTCCACGTA-3 ') was used.

A 212 base pair (bp) sized band, H109 cDNA (SEQ ID NOs: 284-495 bp) was cut out from the dried gel. After elution of H109 cDNA by heating for 15 minutes, PCR was performed under the same primers and the same conditions as above except that [α-35S] labeled dATP (1200 Ci / mmole) and 20 μM dNTP were not used. Amplified.

2-10: TRG  13

H-T11C fixed primer having the nucleotide sequence set forth in SEQ ID NO: 39 (5'-AAGCTTTTTTTTTTTC-3 ', RNAimage kit, Genhunter, Cor., MA, USA) and H having the nucleotide sequence set forth in SEQ ID NO: 40 PCR was performed by the same procedure as in 2-1, except that -AP11 primer (5'-AAGCTTCGGGTAA-3 ') was used.

A 232 base pair (bp) sized band, H119 cDNA (SEQ ID NO: 37, 1004-1235 bp) was cut out from the dried gel. After eluting H119 cDNA by heating for 15 minutes, PCR was performed under the same primers and the same conditions as above except that [α-35S] labeled dATP (1200 Ci / mmole) and 20 μM dNTP were not used. Amplified.

2-11: TRG  14

H-T11G fixed primer having the nucleotide sequence set forth in SEQ ID NO: 43 (5'-AAGCTTTTTTTTTTTG-3 ', RNAimage kit, Genhunter, Cor., MA, USA) and H having the nucleotide sequence set forth in SEQ ID NO: 44 PCR was performed by the same procedure as in 2-1 except that the AP20 primer (5'-AAGCTTGTTGTGC-3 ') was used.

A band of 195 base pairs (bp) in size, H201 cDNA (SEQ ID NO: 902-1096 bp) was cut out from the dried gel. After heating for 15 minutes to elute H201 cDNA, PCR was performed under the same primers and the same conditions as above except that [α-35S] labeled dATP (1200 Ci / mmole) and 20 μM dNTP were not used. Amplified.

2-12: TRG  15

H-T11A fixed primer having the nucleotide sequence set forth in SEQ ID NO: 47 (5'-AAGCTTTTTTTTTTTA-3 ', RNAimage kit, Genhunter, Cor., MA, USA) and H having the nucleotide sequence set forth in SEQ ID NO: 48 PCR was performed by the same procedure as in 2-1 except that the AP15 primer (5'-AAGCTTACGCAAC-3 ') was used.

A 252 base pair (bp) sized band, H151 cDNA (SEQ ID NO: 848-1099 bp) was cut out from the dried gel. After heating for 15 minutes to elute H151 cDNA, PCR was performed under the same primers and the same conditions as above except that [α-35S] labeled dATP (1200 Ci / mmole) and 20 μM dNTP were not used. Amplified.

2-13: TRG  16

H-T11C fixed primer (5'-AAGCTTTTTTTTTTTC-3 ', RNAimage kit, Genhunter, Cor., MA, USA) having the nucleotide sequence set forth in SEQ ID NO: 51 and H having the nucleotide sequence set forth in SEQ ID NO: 52 PCR was performed in the same manner as in 2-1, except that -AP13 primer (5'-AAGCTTCGGCATA-3 ') was used.

A 227 base pair (bp) sized band, H132 cDNA (813-1039 bp of SEQ ID NO: 49) was cut out from the dried gel. After heating for 15 minutes to elute H132 cDNA, PCR was performed under the same primers and the same conditions as above except that [α-35S] labeled dATP (1200 Ci / mmole) and 20 μM dNTP were not used. Amplified.

2-14: TRG  17

H-T11G fixed primer having the nucleotide sequence set forth in SEQ ID NO: 55 (5'-AAGCTTTTTTTTTTTG-3 ', RNAimage kit, Genhunter, Cor., MA, USA) and H having the nucleotide sequence set forth in SEQ ID NO: 56 PCR was performed in the same manner as in 2-1, except that -AP14 primer (5'-AAGCTTGGAGCTT-3 ') was used.

A 185 base pair (bp) sized band, H141 cDNA (SEQ ID NOs: 235-419 bp) was cut out from the dried gel. After eluting H141 cDNA by heating for 15 minutes, PCR was performed under the same primers and the same conditions as above except that [α-35S] labeled dATP (1200 Ci / mmole) and 20 μM dNTP were not used. Amplified.

2-15: TRG  18

H-T11C fixed primer having the nucleotide sequence set forth in SEQ ID NO: 59 (5'-AAGCTTTTTTTTTTTC-3 ', RNAimage kit, Genhunter, Cor., MA, USA) and H having the nucleotide sequence set forth in SEQ ID NO: 60 PCR was carried out in the same manner as in 2-1, except that -AP18 primer (5'-AAGCTTAGAGGCA-3 ') was used.

A 227 base pair (bp) sized band, H181 cDNA (SEQ ID NO: 902-1128 bp) was cut out from the dried gel. After heating for 15 minutes to elute H181 cDNA, PCR was performed under the same primers and the same conditions as above, except that [α-35S] labeled dATP (1200 Ci / mmole) and 20 μM dNTP were not used. Amplified.

2-16: TRG  20

H-T11A fixed primer (5'-AAGCTTTTTTTTTTTA-3 ', RNAimage kit, Genhunter, Cor., MA, USA) having the nucleotide sequence set forth in SEQ ID NO: 63 and H having the nucleotide sequence set forth in SEQ ID NO: 64 PCR was performed in the same manner as in 2-1, except that -AP13 primer (5'-AAGCTTCGGCATA-3 ') was used.

A 186 base pair (bp) size band, H134 cDNA (SEQ ID NOs: 232-417 bp) was cut out from the dried gel. After heating for 15 minutes to elute H134 cDNA, PCR was performed under the same primers and the same conditions as above except that [α-35S] labeled dATP (1200 Ci / mmole) and 20 μM dNTP were not used. Amplified.

Example 3 @ Cloning

Re-amplified H20-121 obtained above; H93-811; H117-321; H38-211; H38-621; H96; H94; H42; H109; H119; H201; H151; H132; H141; H181; Alternatively, the H134 PCR product was inserted into the pGEM-T EASY vector according to the manufacturer's method using the TA Cloning System (Promega, USA).

(Step 1) Connection reaction

Reamplified H20-121 obtained in Example 2; H93-811; H117-321; H38-211; H38-621; H96; H94; H42; H109; H119; H201; H151; H132; H141; H181 or H134 PCR product 2 μl, pGEM-T EASY vector (50 ng) 1, 4 DNA linker 10X buffer 1 and T4 DNA ligase 1 μL of 3 weiss units / μL; T4 DNA ligase, Promega) was placed in a 0.5 μL test tube, and distilled water was added so that the final volume was 10 μL. The ligation reaction mixture was incubated at 14 ° C. overnight.

(Step 2) TA cloning transformation

Escherichia coli JM109 (Promega, WI, USA) was subjected to an optical density value of about 0.3 to 0.6 at 600 nm in 10 ml of LB broth (10 g of bacto-tryptone, 5 g of bacto-yeast extract, 5 g of NaCl). Incubate until. The culture mixture was left on ice for about 10 minutes, then left at 4 ° C. for 10 minutes, and then centrifuged at 4000 rpm for 10 minutes at 4 ° C. to discard the supernatant and cells were collected. The collected cell pellets were exposed to 10 ml of ice cold 0.1 M CaCl ₂ for about 30 minutes to 1 hour to prepare competent cells. The resultant was centrifuged again at 4000 rpm for 10 minutes at 4 ° C. to discard the supernatant, and the cells were collected and suspended in 2 ml of ice-cold 0.1 M CaCl ₂ .

200 μl of competent cell suspension was transferred to a new microfuge tube, and 2 μl of the ligation reaction product prepared in step 1 was added thereto. The mixture was incubated for 90 seconds in a 42 ° C. water bath and then quenched at 0 ° C. Here, SOC medium (2.0 g of bacto-tryptone, 0.5 g of bacto-yeast extract, 1 ml of 1 M NaCl, 0.25 ml of 1 M KCI, 97 ml of TDW, 2 ml of 2 M Mg ^{2 +} 1 ml, 2 ml of glucose) 800 μl was added and the mixture was incubated for 45 minutes in a rotary shake incubator at 37 ° C. at 220 rpm.

25 μl of X-gal (stored in 40 mg / ml dimethylformamide) was spread on a glass rod in an ampicillin-added LB plate, previously placed in a 37 ° C. incubator, and then 25 μl of transformed cells were added thereto. Diffused into a glass rod again and incubated overnight at 37 ℃. Three to four white colonies formed after incubation were selected and each selected clones were planted in LB plates to which ampicillin was added. Colonies of which insertion has been identified, for example, transformed E. coli JM109 / H20-121; JM109 / H93-811; JM109 / H117-321; JM109 / H38-211; JM109 / H38-621; JM109 / H96; JM109 / H94; JM109 / H42; JM109 / H109; JM109 / H119; JM109 / H201; JM109 / H151; JM109 / H132; JM109 / H141; JM109 / H181; Or JM109 / H134 to select 10 ml of terrific broth (900 ml of TDW, 12 g of bacto-tryptone, 24 g of bacto-yeast extract, 4 ml of glycerol, 0.17 M KH ₂ PO ₄ , 0.72 N K ₂ HPO ₄ 100 ml).

Example 4 Separation of Recombination Plasmid DNA

Wizard Plus Mini program repseu DNA purification kit (Wizard ^TM Plus Minipreps DNA Purificatin Kit, Promega, USA) to H20-121 from E. coli transformed according to the manufacturer's instructions using; H93-811; H117-321; H38-211; H38-621; H96; H94; H42; H109; H119; H201; H151; H132; H141; H181; Or H134 plasmid DNA was isolated.

A portion of the isolated plasmid DNA was treated with ECo RI enzyme, followed by electrophoresis on a 2% gel, followed by H20-121; H93-811; H117-321; H38-211; H38-621; H96; It was confirmed that H94; H42; H109; H119; H201; H151; H132; H141; H181; or H134 subsequences were inserted into the plasmid.

Example 5 DNA DNA Base Sequence Analysis

5-1; TRG  3

After amplifying the H20-121 PCR product obtained in Example 2 according to the conventional method, the cloned and re-amplified H20-121 PCR fragment was prepared using a sequencer version 2.0 DNA sequencing kit (United States). Biochemical, Cleveland, OH, USA were used to sequence analysis according to the dideoxy chain termination.

The base sequence of the DNA corresponds to nucleotide numbers 1342 to 1599 of SEQ ID NO: 1, and is referred to as "H20-121" in the present invention.

The 258 bp cDNA fragment obtained above, i.e., H20-121, was subjected to differential development reverse transcription polymerase chain reaction (DDRT-PCR) using 5 'random primers H-AP20 and 3' H-T11A fixed primers, followed by electrophoresis. Confirmed.

As shown in FIG. 1A, gene expression by DD in normal cervical tissue, metastatic lymph node tissue and CUMC-6 cells was found to be different. As can be seen in Figure 1a, 258 bp cDNA fragment H20-121 was expressed in cervical cancer, metastatic lymph node tissue and CUMC-6 cancer cells, but the expression was very weak in normal tissues.

5-2; TRG  4

After amplifying the H93-811 PCR product obtained in Example 2 according to a conventional method, the cloned and re-amplified H93-811 PCR fragment was prepared using a sequencer DNA 2.0 sequencing kit; United States Biochemical, Cleveland, OH, USA) was used to sequence analysis according to dideoxy chain termination (dideoxy chain termination).

The “base” sequence of the DNA corresponds to “nucleotides” of SEQ ID NO: 5 “2086 to 2478” and “H93-811” in the present invention.

Using the 5 'random primers H-AP9 and 3 H-T11G fixed primers, the 393bp cDNA fragment obtained above, ie, H93-811, was subjected to differential development reverse transcription polymerase chain reaction (DDRT-PCR) and electrophoresis. Confirmed.

As shown in Figure 1b, gene expression by DD in normal cervical tissue, metastatic lymph node tissue, and CUMC-6 cells was found to be different. As shown in Figure 1b, H93-811, a 393bp bp cDNA fragment, was expressed in cervical cancer, metastatic lymph node tissue, and CUMC-6 cancer cells, but was very weak in normal tissue.

5-3; TRG  5

After amplifying the H117-321 PCR product obtained in Example 2 according to a conventional method, the cloned and re-amplified H117-321 PCR fragment was prepared using a sequencer version 2.0 DNA sequencing kit (United States). Biochemical, Cleveland, OH, USA) was used to sequence analysis according to dideoxy chain termination (dideoxy chain termination).

The base sequence of the DNA corresponds to nucleotide numbers 933 to 1224 of SEQ ID NO: 9, and is referred to as "H117-321" in the present invention.

The 292 bp cDNA fragment obtained above, H117-321, using 5 'random primers H-AP11 and 3' H-T11C fixed primers, was subjected to differential reverse transcription polymerase chain reaction (DDRT-PCR) and subjected to electrophoresis. Confirmed.

As shown in FIG. 1C, gene expression by DD in normal cervical tissue, metastatic lymph node tissue and CUMC-6 cells was found to be different. As can be seen in Figure 1c, the 292 bp cDNA fragment H117-321 was expressed in cervical cancer, metastatic lymph node tissue and CUMC-6 cancer cells, but the expression was very weak in normal tissues.

5-4; TRG  6

After amplifying the H38-211 PCR product obtained in Example 2 according to a conventional method, the cloned and re-amplified H38-211 PCR fragment was prepared using a sequencer version 2.0 DNA sequencing kit (United States). Biochemical, Cleveland, OH, USA) was used to sequence analysis according to dideoxy chain termination (dideoxy chain termination).

The base sequence of the DNA corresponds to nucleotide numbers 2823 to 3133 of SEQ ID NO: 13, referred to as "H38-211" in the present invention. The 311 bp cDNA fragment obtained above, H38-211, using 5 'random primers H-AP38 and 3' H-T11A fixed primers was subjected to differential reverse reverse transcription polymerase chain reaction (DDRT-PCR) and subjected to electrophoresis. Confirmed.

As shown in FIG. 1D, gene expression by DD in normal cervical tissue, metastatic lymph node tissue and CUMC-6 cells was found to be different. As can be seen in Figure 1d, the 311 bp cDNA fragment H38-211 was expressed in cervical cancer, metastatic lymph node tissue and CUMC-6 cancer cells, but the expression was very weak in normal tissues.

5-5; TRG  7

After amplifying the H38-621 PCR product obtained in Example 2 according to a conventional method, the cloned and re-amplified H38-621 PCR fragment was prepared using a sequencer version 2.0 DNA sequencing kit (United States). Biochemical, Cleveland, OH, USA) was used to sequence analysis according to dideoxy chain termination (dideoxy chain termination).

The base sequence of the DNA corresponds to nucleotide numbers 1404 to 1695 of SEQ ID NO: 17, and is referred to as "H38-621" in the present invention.

The 292 bp cDNA fragments obtained above, H38-621, using 5 'random primers H-AP38 and 3' H-T11G immobilized primers were subjected to differential reverse reverse transcription polymerase chain reaction (DDRT-PCR) and subjected to electrophoresis. Confirmed.

As shown in FIG. 1E, gene expression by DD was found to be different in normal cervical tissue, metastatic lymph node tissue and CUMC-6 cells. As can be seen in Figure 1e, 292 bp cDNA fragment H38-621 was expressed in cervical cancer, metastatic lymph node tissue and CUMC-6 cancer cells, but the expression was very weak in normal tissues.

5-6; TRG  9

After amplifying the H96 PCR product obtained in Example 2 according to a conventional method, the cloned and re-amplified H96 PCR fragment was subjected to a sequence version 2.0 DNA sequencing kit (United States Biochemical, Cleveland, OH, USA) was used to sequence analysis according to dideoxy chain termination (dideoxy chain termination).

The base sequence of the DNA corresponds to nucleotide numbers 1225 to 1499 of SEQ ID NO: 21, and is referred to as "H96" in the present invention.

The 275 bp cDNA fragment obtained above, ie H96, using 5 'random primers H-AP9 and 3' H-T11A fixed primers was subjected to differential development reverse transcriptase polymerase chain reaction (DDRT-PCR) and confirmed by electrophoresis It was.

As shown in FIG. 1F, gene expression by DD in normal cervical tissue, metastatic lymph node tissue and CUMC-6 cells was found to be different. As can be seen in FIG. 1F, H96, a 275 bp cDNA fragment, was expressed in cervical cancer, metastatic lymph node tissue and CUMC-6 cancer cells, but was very weak in normal tissue.

5-7; TRG10

After amplifying the H94 PCR product obtained in Example 2 according to a conventional method, the cloned and re-amplified H94 PCR fragments were prepared using a Sequenase version 2.0 DNA Sequencing kit (United States Biochemical, Cleveland, OH, USA) was used to sequence analysis according to dideoxy chain termination (dideoxy chain termination).

The base sequence of the DNA corresponds to nucleotide numbers 3528 to 3879 of SEQ ID NO: 25, and is referred to as "H94" in the present invention.

The 352 bp cDNA fragment obtained above, ie, H94, using 5 'random primers H-AP9 and 3' H-T11A fixed primers, was differentially developed by reverse transcription polymerase chain reaction (DDRT-PCR) and confirmed by electrophoresis. .

As shown in FIG. 1G, gene expression by DD in normal cervical tissue, metastatic lymph node tissue and CUMC-6 cells was found to be different. As can be seen in Figure 1g, 352 bp cDNA fragment H94 was expressed in cervical cancer, metastatic lymph node tissue and CUMC-6 cancer cells, but weak in normal tissues.

5-8; TRG11

After amplifying the H42 PCR product obtained in Example 2 according to a conventional method, the cloned and re-amplified H42 PCR fragments were prepared using a Sequenase version 2.0 DNA Sequencing kit (United States Biochemical, Cleveland, OH, USA) was used to sequence analysis according to dideoxy chain termination (dideoxy chain termination).

The base sequence of the DNA corresponds to nucleotide numbers 83 to 229 of SEQ ID NO: 29, which is referred to as "H42" in the present invention.

The 147 bp cDNA fragment obtained above, i.e., H42, using 5 'random primers H-AP4 and 3' H-T11C fixed primers was subjected to differential development reverse transcriptase polymerase chain reaction (DDRT-PCR) and confirmed by electrophoresis. .

As shown in FIG. 1H, gene expression by DD in normal cervical tissue, metastatic lymph node tissue and CUMC-6 cells was found to be different. As can be seen in FIG. 1H, H42, a 147 bp cDNA fragment, was expressed in cervical cancer, metastatic lymph node tissue and CUMC-6 cancer cells, but was weak in normal tissue.

5-9; TRG  12

After amplifying the H109 PCR product obtained in Example 2 according to a conventional method, the cloned and re-amplified H109 PCR fragment was subjected to a sequence version 2.0 DNA sequencing kit (United States Biochemical, Cleveland, OH, USA) was used for sequencing according to dideoxy chain termination.

The base sequence of the DNA corresponds to nucleotide numbers 284 to 495 of SEQ ID NO: 33, which is referred to as "H109" in the present invention.

The 212 bp cDNA fragment obtained above, i.e., H109, using 5 'random primers H-AP10 and 3' H-T11G fixed primers, was subjected to differential development reverse transcription polymerase chain reaction (DDRT-PCR) and confirmed by electrophoresis. .

As shown in FIG. 1I, gene expression by DD in normal cervical tissue, metastatic lymph node tissue and CUMC-6 cells was found to be different. As can be seen in FIG. 1i, H12, a 212 bp cDNA fragment, was expressed in cervical cancer, metastatic lymph node tissue and CUMC-6 cancer cells, but was very weak in normal tissue.

5-10; TRG  13

After amplifying the H119 PCR product obtained in Example 2 according to a conventional method, the cloned and re-amplified H119 PCR fragment was subjected to a sequence version 2.0 DNA sequencing kit (United States Biochemical, Cleveland, OH, USA) was used to sequence analysis according to dideoxy chain termination (dideoxy chain termination).

The base sequence of the DNA corresponds to nucleotide numbers 1004 to 1235 of SEQ ID NO: 37, and is referred to as "H119" in the present invention.

The 232 bp cDNA fragment obtained above, ie, H119, using 5 'random primers H-AP11 and 3' H-T11C fixed primers was subjected to differential reverse reverse transcription polymerase chain reaction (DDRT-PCR) and confirmed by electrophoresis. .

As shown in FIG. 1J, gene expression by DD in normal cervical tissue, metastatic lymph node tissue and CUMC-6 cells was found to be different. As can be seen in Figure 1j, the 232 bp cDNA fragment H119 was expressed in cervical cancer, metastatic lymph node tissue and CUMC-6 cancer cells, but very weak in normal tissues.

5-11; TRG  14

After amplifying the H201 PCR product obtained in Example 2 according to a conventional method, the cloned and re-amplified H201 PCR fragment was prepared using a sequencer version 2.0 DNA sequencing kit (United States Biochemical, Cleveland, OH, USA) was used to sequence analysis according to dideoxy chain termination (dideoxy chain termination).

The base sequence of the DNA corresponds to nucleotide numbers 902 to 1096 of SEQ ID NO: 41, and is referred to as "H201" in the present invention.

The 195 bp cDNA fragment obtained above, H201, using 5 'random primers H-AP20 and 3' H-T11G fixed primers, was subjected to differential development reverse transcription polymerase chain reaction (DDRT-PCR) and confirmed by electrophoresis. .

As shown in FIG. 1K, gene expression by DD in normal cervical tissue, metastatic lymph node tissue and CUMC-6 cells was found to be different. As can be seen in FIG. 1K, H201, a 195 bp cDNA fragment, was expressed in cervical cancer, metastatic lymph node tissue, and CUMC-6 cancer cells, but was very weak in normal tissue.

5-12; TRG  15

After amplifying the H151 PCR product obtained in Example 2 according to a conventional method, the cloned and re-amplified H151 PCR fragments were prepared using a Sequenase version 2.0 DNA Sequencing kit (United States Biochemical, Cleveland, OH, USA) was used to sequence analysis according to dideoxy chain termination (dideoxy chain termination).

The base sequence of the DNA corresponds to nucleotide numbers 848 to 1099 of SEQ ID NO: 45, and is referred to as "H151" in the present invention.

The 252 bp cDNA fragment obtained above, H151, using 5 'random primers H-AP15 and 3' H-T11A fixed primers, was subjected to differential reverse reverse transcriptase polymerase chain reaction (DDRT-PCR) and confirmed by electrophoresis. .

As shown in FIG. 1 (l), gene expression by DD in normal cervical tissue, metastatic lymph node tissue and CUMC-6 cells was found to be different. As can be seen in Figure 1 (l), 252 bp cDNA fragment H151 was expressed in cervical cancer, metastatic lymph node tissue and CUMC-6 cancer cells, but the expression was very weak in normal tissue.

5-13; TRG  16

After amplifying the H132 PCR product obtained in Example 2 according to a conventional method, the cloned and re-amplified H132 PCR fragment was subjected to a sequence version 2.0 DNA sequencing kit (United States Biochemical, Cleveland, OH, USA) was used for sequencing according to dideoxy chain termination.

The base sequence of the DNA corresponds to nucleotides 813 to 1039 of SEQ ID NO: 49, and is referred to as "H132" in the present invention.

The 227 bp cDNA fragment obtained above, i.e., H132, using 5 'random primers H-AP13 and 3' H-T11C fixed primers was differentially developed by reverse transcription polymerase chain reaction (DDRT-PCR) and confirmed by electrophoresis. .

As shown in FIG. 1M, gene expression by DD in normal cervical tissue, metastatic lymph node tissue and CUMC-6 cells was found to be different. As can be seen in Figure 1m, H227, a 227 bp cDNA fragment was expressed in cervical cancer, metastatic lymph node tissue and CUMC-6 cancer cells, but the expression was very weak in normal tissues.

5-14; TRG  17

 After amplifying the H141 PCR product obtained in Example 2 according to a conventional method, the cloned and re-amplified H141 PCR fragments were prepared using a Sequenase version 2.0 DNA Sequencing kit (United States Biochemical, Cleveland, OH, USA) was used to sequence analysis according to dideoxy chain termination (dideoxy chain termination).

The base sequence of the DNA corresponds to nucleotide numbers 235 to 419 of SEQ ID NO: 53, and was referred to as "H141" in the present invention.

The 185 bp cDNA fragment obtained above, i.e., H141, using 5 'random primers H-AP14 and 3' H-T11G fixed primers was subjected to differential development reverse transcription polymerase chain reaction (DDRT-PCR) and confirmed by electrophoresis. .

As shown in FIG. 1N, gene expression by DD in normal cervical tissue, metastatic lymph node tissue and CUMC-6 cells was found to be different. As can be seen in FIG. 1N, H141, a 185 bp cDNA fragment, was expressed in cervical cancer, metastatic lymph node tissue and CUMC-6 cancer cells, but was very weak in normal tissue.

5-15; TRG  18

After amplifying the H181 PCR product obtained in Example 2 according to a conventional method, the cloned and re-amplified H181 PCR fragment was prepared using a Sequenase version 2.0 DNA Sequencing kit (United States Biochemical, Cleveland, OH, USA) was used to sequence analysis according to dideoxy chain termination (dideoxy chain termination).

The base sequence of the DNA corresponds to nucleotide numbers 902 to 1128 of SEQ ID NO: 57, and is referred to as "H181" in the present invention.

The 227 bp cDNA fragment obtained above, ie, H181, using 5 'random primers H-AP18 and 3' H-T11C fixed primers was subjected to differential development reverse transcriptase polymerase chain reaction (DDRT-PCR) and confirmed by electrophoresis. .

As shown in FIG. 1 (o), gene expression by DD in normal cervical tissue, metastatic lymph node tissue and CUMC-6 cells was found to be different. As can be seen in FIG. 1 (o), H227, a 227 bp cDNA fragment, was expressed in cervical cancer, metastatic lymph node tissue and CUMC-6 cancer cells, but was very weak in normal tissue.

5-16; TRG  20

After amplifying the H134 PCR product obtained in Example 2 according to a conventional method, the cloned and re-amplified H134 PCR fragment was subjected to a sequence version 2.0 DNA sequencing kit (United States Biochemical, Cleveland, OH, USA) was used to sequence analysis according to dideoxy chain termination (dideoxy chain termination).

The base sequence of the DNA corresponds to nucleotide numbers 232 to 417 of SEQ ID NO: 61, and is referred to as "H134" in the present invention.

The 186 bp cDNA fragment obtained above, i.e., H134, using 5 'random primers H-AP13 and 3' H-T11A fixed primers, was differentially developed reverse transcriptase polymerase chain reaction (DDRT-PCR) and confirmed by electrophoresis. .

As shown in FIG. 1P, gene expression by DD in normal cervical tissue, metastatic lymph node tissue and CUMC-6 cells was found to be different. As can be seen in Figure 1p, the 186 bp cDNA fragment H134 was expressed in cervical cancer, metastatic lymph node tissue and CUMC-6 cancer cells, but very weak in normal tissues.

Example 6 Analysis of total cDNA sequence of TRG source cancer gene

6-1; TRG  3

Bacteriophage λgt11 human lung fetal fibroblast cDNA library using 32P-labeled H20-121 as a probe (Miki, T. et. al . , Gene 83: 137-146, 1989). From the human lung fetal fibroblast cDNA library, a full TRG3 cDNA clone with 1703 bp inserted into the pCEV-LAC vector was obtained and registered with GeneBank, US, Dec. No. AY189688 (Dec. 2, 2002). Scheduled date: April 8, 2005).

TRG3 clones inserted into λpCEV vectors from phage were isolated in the form of ampicillin-resistant pCEV-LAC phagemid vectors by Not I cleavage (Miki, T. et. al . , Gene 83: 137-146, 1989). The pCEV-LAC vector containing the TRG3 gene was linked with T4 DNA ligase to prepare TRG3 plasmid DNA and transformed into E. coli DH5α with the linked clone.

The entire nucleotide sequence of TRG3 consisting of 1703 bp is shown in SEQ ID NO: 1.

In the nucleotide sequence of SEQ ID NO: 1, the entire open reading frame of the proto-oncogene of the present invention corresponds to nucleotides 113 to 1522, predicted to encode a protein consisting of the 469 amino acids of SEQ ID NO: 2 do.

6-2; TRG  4

Bacteriophage λgt11 human lung fetal fibroblast cDNA library using ³² P-labeled H93-811 as a probe (Miki, T. et. 　 al . , Gene 83: 137-146, 1989). From the human lung fetal fibroblast cDNA library, a full TRG4 cDNA clone with 2576 bp inserted into the pCEV-LAC vector was obtained and registered with GeneBank of the United States on December 2, 2002 under the accession number AY189690. Scheduled date: April 8, 2005). TRG4 clones inserted into λpCEV vectors from phage were isolated in the form of ampicillin resistant pCEV-LAC phagemid vectors by Not I cleavage (Miki, T. et. al ., Gene 83: 137-146, 1989). The pCEV-LAC vector containing the TRG4 gene was linked with T4 DNA ligase to prepare TRG4 plasmid DNA and transformed with E. coli DH5α with the linked clone. The entire nucleotide sequence of TRG4 consisting of 2576 bp is shown in SEQ ID NO: 5. In the nucleotide sequence of SEQ ID NO: 5, the full open reading frame of the proto-oncogene of the present invention corresponds to nucleotides 87 to 482 and is predicted to encode a protein consisting of 131 amino acids of SEQ ID NO: 6 do.

6-3; TRG  5

Bacteriophage λgt11 human lung fetal fibroblast cDNA library using 32P-labeled H117-321 as a probe (Miki, T. et. al . , Gene 83: 137-146, 1989). From the human lung fetal fibroblast cDNA library, a full TRG5 cDNA clone with 1334 bp inserted into the pCEV-LAC vector was obtained and registered with GeneBank, US, Dec. No. AY189689 as of December 2, 2002. Scheduled date: April 8, 2005).

TRG5 clone inserted into λpCEV vector from phage was isolated in the form of ampicillin resistant pCEV-LAC phagemid vector by Not I cleavage (Miki, T. et. al . , Gene 83: 137-146, 1989). The pCEV-LAC vector containing the TRG5 gene was linked with a T4 DNA ligase to prepare TRG5 plasmid DNA and transformed into E. coli DH5α with the linked clone. The entire nucleotide sequence of TRG5 consisting of 1336 bp is shown in SEQ ID NO: 9.

In the nucleotide sequence of SEQ ID NO: 9, the entire open reading frame of the proto-oncogene of the present invention corresponds to nucleotides 88 to 1092, predicted to encode a protein consisting of 334 amino acids of SEQ ID NO: 10. do.

6-4; TRG  6

Bacteriophage λgt11 human lung fetal fibroblast cDNA library using 32P-labeled H38-211 as a probe (Miki, T. et. al . , Gene 83: 137-146, 1989). From the human lung fetal fibroblast cDNA library, a full TRG6 cDNA clone with 3309 bp inserted into the pCEV-LAC vector was obtained and registered with GeneBank of the United States as Gen. Scheduled date: April 8, 2005. TRG6 clones inserted into λpCEV vectors from phage were isolated in the form of ampicillin-resistant pCEV-LAC phagemid vectors by Not I cleavage (Miki, T. et. al . , Gene 83: 137-146, 1989).

The pCEV-LAC vector containing the TRG6 gene was linked with T4 DNA ligase to prepare TRG6 plasmid DNA and transformed with E. coli DH5α with the linked clone.

The entire nucleotide sequence of TRG6 consisting of 3309 bp is shown in SEQ ID NO: 13.

In the nucleotide sequence of SEQ ID NO: 13, the full open reading frame of the proto-oncogene of the present invention corresponds to nucleotides 233 to 481, predicted to encode a protein consisting of 82 amino acids of SEQ ID NO: 14. do.

6-5; TRG  7

Bacteriophage λgt11 human lung fetal fibroblast cDNA library using 32P-labeled H38-621 as a probe (Miki, T. et. al . , Gene 83: 137-146, 1989). From the human lung fetal fibroblast cDNA library, a full TRG7 cDNA clone with 1778 bp inserted into the pCEV-LAC vector was obtained and registered with GeneBank of the United States on December 5, 2002 under the accession no. Scheduled date: April 8, 2005). TRG7 clones inserted into λpCEV vectors from phage were isolated in the form of ampicillin resistant pCEV-LAC phagemid vectors by Not I cleavage (Miki, T. et. al . , Gene 83: 137-146, 1989).

The pCEV-LAC vector containing the TRG7 gene was linked with T4 DNA ligase to prepare TRG7 plasmid DNA and transformed into E. coli DH5α with the linked clone. The entire nucleotide sequence of TRG7 consisting of 1778 bp is shown in SEQ ID NO: 17.

In the nucleotide sequence of SEQ ID NO: 17, the entire open reading frame of the proto-oncogene of the present invention corresponds to nucleotides 42 to 1422, predicted to encode a protein consisting of 175 amino acids of SEQ ID NO: 18. do.

6-6; TRG  9

Bacteriophage λgt11 human lung fetal fibroblast cDNA library using 32P-labeled H96 as a probe (Miki, T. et. al . , Gene 83: 137-146, 1989). From the human lung fetal fibroblast cDNA library, a full TRG9 cDNA clone with 1582 bp inserted into the pCEV-LAC vector was obtained and registered with GeneBank, US Patent No. AY272044, dated April 9, 2003 (public publication). Scheduled date: March 31, 2005).

TRG9 clones inserted into λpCEV vectors from phage were isolated in the form of ampicillin-resistant pCEV-LAC phagemid vectors by Not I cleavage (Miki, T. et. al . , Gene 83: 137-146, 1989). The pCEV-LAC vector containing the TRG9 gene was linked with T4 DNA ligase to prepare TRG9 plasmid DNA and transformed with E. coli DH5α with the linked clone.

The entire nucleotide sequence of TRG9 consisting of 1582 bp is shown in SEQ ID NO: 21.

In the nucleotide sequence of SEQ ID NO: 21, the entire open reading frame of the proto-oncogene of the present invention corresponds to nucleotides 17 to 1576, predicted to encode a protein consisting of 519 amino acids of SEQ ID NO: 22. do.

6-7; TRG10

Bacteriophage λgt11 human lung fetal fibroblast cDNA library using 32P-labeled H94 as a probe (Miki, T. et. al . , Gene 83: 137-146, 1989). From the human lung fetal fibroblast cDNA library, a full TRG10 cDNA clone with 3979 bp inserted into the pCEV-LAC vector was obtained and registered with GeneBank, US Patent No. AY277593 as of April 12, 2003 (public publication). Scheduled date: March 31, 2005).

TRG10 clones inserted into λpCEV vectors from phage were isolated in the form of ampicillin resistant pCEV-LAC phagemid vectors by Not I cleavage (Miki, T. et. al . , Gene 83 : 137-146, 1989).

The pCEV-LAC vector containing the TRG10 gene was linked with T4 DNA ligase to prepare TRG10 plasmid DNA and transformed with E. coli DH5α with the linked clone.

The entire nucleotide sequence of TRG10 consisting of 3979 bp is shown in SEQ ID NO: 25.

In the nucleotide sequence of SEQ ID NO: 25, the entire open reading frame of the proto-oncogene of the present invention corresponds to nucleotides 1100 to 1270, predicted to encode a protein consisting of 56 amino acids of SEQ ID NO: 26. do.

6-8; TRG11

Bacteriophage λgt11 human lung fetal fibroblast cDNA library using 32P-labeled H42 as a probe (Miki, T. et. al . , Gene 83: 137-146, 1989). From the human lung fetal fibroblast cDNA library, a full TRG11 cDNA clone with 235 bp inserted into the pCEV-LAC vector was obtained and registered with GeneBank of the United States on April 13, 2003 under accession number AY277594. Scheduled date: March 31, 2005).

TRG11 clones inserted into λpCEV vectors from phage were isolated in the form of ampicillin resistant pCEV-LAC phagemid vectors by Not I cleavage (Miki, T. et. al . , Gene 83: 137-146, 1989).

The pCEV-LAC vector containing the TRG11 gene was linked with T4 DNA ligase to prepare TRG11 plasmid DNA, and transformed into E. coli DH5α with the linked clone.

The entire nucleotide sequence of TRG11 consisting of 235 bp is shown in SEQ ID NO: 29.

In the nucleotide sequence of SEQ ID NO: 29, the entire open reading frame of the proto-oncogene of the present invention corresponds to nucleotides 26 to 214, and predicted to encode a protein consisting of 62 amino acids of SEQ ID NO: 30 do.

6-9; TRG  12

Bacteriophage λgt11 human lung fetal fibroblast cDNA library using 32P-labeled H109 as a probe (Miki, T. et. al . , Gene 83: 137-146, 1989). From the human lung fetal fibroblast cDNA library, a full TRG12 cDNA clone with 510 bp inserted into the pCEV-LAC vector was obtained and registered with GeneBank, US Patent No. AY277595, dated April 13, 2003 (public publication). Scheduled date: March 31, 2005).

TRG12 clone inserted into λpCEV vector from phage was isolated in the form of ampicillin resistant pCEV-LAC phagemid vector by Not I cleavage (Miki, T. et. al . , Gene 83: 137-146, 1989).

The pCEV-LAC vector containing the TRG12 gene was linked with T4 DNA ligase to prepare TRG12 plasmid DNA and transformed with E. coli DH5α with the linked clone.

The entire nucleotide sequence of TRG12 consisting of 510 bp is shown in SEQ ID NO: 33.

In the nucleotide sequence of SEQ ID NO: 33, the entire open reading frame of the proto-oncogene of the present invention corresponds to nucleotides 80 to 475, predicted to encode a protein consisting of 131 amino acids of SEQ ID NO: 34. do.

6-10; TRG  13

Bacteriophage λgt11 human lung fetal fibroblast cDNA library using 32P-labeled H119 as a probe (Miki, T. et. al . , Gene 83: 137-146, 1989). From the human lung fetal fibroblast cDNA library, a full TRG13 cDNA clone with 1301 bp inserted into the pCEV-LAC vector was obtained and registered with GeneBank of the United States on April 13, 2003 under the accession number AY277596 (published). Scheduled date: March 31, 2005).

TRG13 clone inserted into λpCEV vector from phage was isolated in the form of ampicillin resistant pCEV-LAC phagemid vector by Not I cleavage (Miki, T. et. al . , Gene 83: 137-146, 1989).

The pCEV-LAC vector containing the TRG13 gene was linked with T4 DNA ligase to prepare TRG13 plasmid DNA and transformed with E. coli DH5α with the linked clone.

The entire nucleotide sequence of TRG13 consisting of 1301 bp is shown in SEQ ID NO: 37.

In the nucleotide sequence of SEQ ID NO: 37, the entire open reading frame of the proto-oncogene of the present invention corresponds to nucleotides 18 to 1193, predicted to encode a protein consisting of 391 amino acids of SEQ ID NO: 38. do.

6-11; TRG  14

Bacteriophage λgt11 human lung fetal fibroblast cDNA library using 32P-labeled H201 as a probe (Miki, T. et. al . , Gene 83: 137-146, 1989). From the human lung fetal fibroblast cDNA library, a full TRG14 cDNA clone with 1206 bp inserted into the pCEV-LAC vector was obtained and registered with GeneBank, US Publication No. AY277597, dated April 13, 2003 (public publication). Scheduled date: March 31, 2005).

TRG14 clone inserted into λpCEV vector from phage was isolated in the form of ampicillin resistant pCEV-LAC phagemid vector by Not I cleavage (Miki, T. et. al . , Gene 83: 137-146, 1989).

The pCEV-LAC vector containing the TRG14 gene was linked with T4 DNA ligase to prepare TRG14 plasmid DNA and transformed with E. coli DH5α with the linked clone.

The entire nucleotide sequence of TRG14 consisting of 1206 bp is shown in SEQ ID NO: 41.

In the nucleotide sequence of SEQ ID NO: 41, the entire open reading frame of the far-oncogene of the present invention corresponds to nucleotides 18 to 1202, predicted to encode a protein consisting of 394 amino acids of SEQ ID 42. do.

6-12; TRG  15

Bacteriophage λgt11 human lung fetal fibroblast cDNA library using 32P-labeled H151 as a probe (Miki, T. et. al . , Gene 83: 137-146, 1989). From the human lung fetal fibroblast cDNA library, a full TRG15 cDNA clone with 1104 bp inserted into the pCEV-LAC vector was obtained and registered with GeneBank, US Patent No. AY277598, dated April 13, 2003 (public publication). Scheduled date: March 31, 2005).

TRG15 clone inserted into λpCEV vector from phage was isolated in the form of ampicillin resistant pCEV-LAC phagemid vector by Not I cleavage (Miki, T. et. al . , Gene 83: 137-146, 1989).

The pCEV-LAC vector containing the TRG15 gene was linked with T4 DNA ligase to prepare TRG15 plasmid DNA and transformed with E. coli DH5α with the linked clone.

The entire nucleotide sequence of TRG15 consisting of 1104 bp is shown in SEQ ID NO: 45.

In the nucleotide sequence of SEQ ID NO: 45, the entire open reading frame of the far-oncogene of the present invention corresponds to nucleotides 1 to 1104 and is predicted to encode a protein consisting of 367 amino acids of SEQ ID NO: 46. do.

6-13; TRG  16

Bacteriophage λgt11 human lung fetal fibroblast cDNA library using 32P-labeled H132 as a probe (Miki, T. et. al . , Gene 83: 137-146, 1989). From the human lung fetal fibroblast cDNA library, a full TRG16 cDNA clone with 1064 bp inserted into the pCEV-LAC vector was obtained and registered with GeneBank, US Patent No. AY277601 as of April 14, 2003 (published). Scheduled date: March 31, 2005).

TRG16 clones inserted into λpCEV vectors from phage were isolated in the form of ampicillin resistant pCEV-LAC phagemid vectors by Not I cleavage (Miki, T. et. al . , Gene 83: 137-146, 1989).

The pCEV-LAC vector containing the TRG16 gene was linked with T4 DNA ligase to prepare TRG16 plasmid DNA and transformed into E. coli DH5α with the linked clone.

The entire nucleotide sequence of TRG16 consisting of 1064 bp is shown in SEQ ID NO: 49.

In the nucleotide sequence of SEQ ID NO: 49, the entire open reading frame of the proto-oncogene of the present invention corresponds to nucleotides 92 to 1064 and is predicted to encode a protein consisting of 324 amino acids of SEQ ID NO: 50. do.

6-14; TRG  17

Bacteriophage λgt11 human lung fetal fibroblast cDNA library using 32P-labeled H141 as a probe (Miki, T. et. al . , Gene 83: 137-146, 1989). From the human lung fetal fibroblast cDNA library, a full TRG17 cDNA clone with 432 bp inserted into the pCEV-LAC vector was obtained and registered with GeneBank, US Patent No. AY277599, dated April 13, 2003 (published). Scheduled date: March 31, 2005). TRG17 clone inserted into λpCEV vector from phage was isolated in the form of ampicillin resistant pCEV-LAC phagemid vector by Not I cleavage (Miki, T. et. al . , Gene 83: 137-146, 1989).

The pCEV-LAC vector containing the TRG17 gene was linked with T4 DNA ligase to prepare TRG17 plasmid DNA, and transformed into E. coli DH5α with the linked clone.

The entire nucleotide sequence of TRG17 consisting of 432 bp is shown in SEQ ID NO: 53.

In the nucleotide sequence of SEQ ID NO: 53, the entire open reading frame of the proto-oncogene of the present invention corresponds to nucleotides 1 to 408, predicted to encode a protein consisting of 135 amino acids of SEQ ID 54: do.

6-15; TRG  18

Bacteriophage λgt11 human lung fetal fibroblast cDNA library using 32P-labeled H181 as a probe (Miki, T. et. al . , Gene 83: 137-146, 1989). From the human lung fetal fibroblast cDNA library, a full TRG18 cDNA clone with 1141 bp inserted into the pCEV-LAC vector was obtained and registered with GeneBank, US Patent No. AY277600, dated April 13, 2003 (published). Scheduled date: March 31, 2005).

TRG18 clones inserted into λpCEV vectors from phage were isolated in the form of ampicillin resistant pCEV-LAC phagemid vectors by Not I cleavage (Miki, T. et. al . , Gene 83: 137-146, 1989).

The pCEV-LAC vector containing the TRG18 gene was linked with T4 DNA ligase to prepare TRG18 plasmid DNA, and transformed into E. coli DH5α with the linked clone.

The entire nucleotide sequence of TRG18 consisting of 1141 bp is shown in SEQ ID NO: 57.

In the nucleotide sequence of SEQ ID NO: 57, the entire open reading frame of the far-oncogene of the present invention corresponds to nucleotides 20 to 1141 and is predicted to encode a protein consisting of 373 amino acids of SEQ ID NO: 58. do.

6-16; TRG  20

Bacteriophage λgt11 human lung fetal fibroblast cDNA library using 32P-labeled H134 as a probe (Miki, T. et. al . , Gene 83: 137-146, 1989). From the human lung fetal fibroblast cDNA library, a full TRG20 cDNA clone with 449 bp inserted into the pCEV-LAC vector was obtained and registered with GeneBank of the United States as Gen. No. AY453397 dated October 29, 2003 ( Scheduled to be released: March 31, 2005).

TRG20 clones inserted into λpCEV vectors from phage were isolated in the form of ampicillin resistant pCEV-LAC phagemid vectors by Not I cleavage (Miki, T. et. al . , Gene 83: 137-146, 1989).

The pCEV-LAC vector containing the TRG20 gene was linked with T4 DNA ligase to prepare TRG20 plasmid DNA and transformed with E. coli DH5α with the linked clone.

The entire nucleotide sequence of TRG20 consisting of 449 bp is shown in SEQ ID NO: 61.

In the nucleotide sequence of SEQ ID NO: 61, the entire open reading frame of the proto-oncogene of the present invention corresponds to nucleotides 42 to 449, predicted to encode a protein consisting of 135 amino acids of SEQ ID NO: 62. do.

Example 7 Northern Blot Analysis of TRG Gene in Various Cells

As in Example 1, total RNA was extracted from normal cervical tissue, cervical cancer tissue, cervical cancer metastasis, lymph node tissue, and cervical cancer cell lines CaSki (ATCC CRL 1550) and CUMC-6.

To determine the expression level of TRG genes, 20 μg of denatured total RNA extracted from each tissue and cell line was electrophoresed on 1% formaldehyde agarose gel and then transferred to nylon membrane (Boehringer-Mannheim, Germany). The blots were hybridized with ³² P-labeled random primed TRG whole cDNA probes prepared using the Rediprime II random prime labeling system (Amersham, UK). Northern blot analysis was repeated twice, and the results were quantified using densitometry and normalized to β-actin.

Figure 2a shows the northern blot analysis results confirming the expression of TRG3 primary cancer genes in normal cervical tissue, cervical cancer tissue, cervical cancer metastatic lymph node tissue and cervical cancer cell lines (CaSki and CUMC-6). As can be seen in Figure 2a, the TRG3 proto-oncogene has increased gene expression in cervical cancer tissues and cervical cancer cell lines CaSki and CUMC-6 cell lines, ie overexpressing a dominant TRG3 mRNA transcript of about 1.7 kb. In FIG. 2A, normal fever is normal cervical tissue, cancer fever is cervical cancer tissue, metastasis fever is cervical cancer metastatic lymph node tissue, and CaSki fever and CUMC-6 fever are Each represents a uterine cancer cell line. 2A bottom shows hybridization of the same sample with β-actin probe to confirm the presence of mRNA.

Figure 3a is a Northern human 12-row multiple tissue (Clontech), brain, heart, rhabdomyo, large intestine, thymus, spleen, kidney, liver, small intestine, placenta, lung and peripheral blood leukocyte tissues to determine whether the expression of TRG3 primary oncogenes The results of the blot analysis are shown. Figure 3b hybridizes the same sample with β-actin probe to confirm the presence of mRNA. As can be seen in Figure 3a, TRG3 mRNA (dominant TRG3 mRNA transcript of about 1.7 kb) is expressed in the heart and normal muscle tissue, but normal brain, colon, thymus, spleen, kidney, liver, small intestine , Weakly expressed in placenta, lung and peripheral blood leukocyte tissues.

Figure 19a shows the results of Northern blot analysis confirming the expression of TRG3 primary oncogenes in human cancer cell lines, HL-60, HeLa, K-562, MOLT-4, Raji, SW480, A549 and G361 (Clontech). 19B shows the presence of mRNA by hybridizing the same sample with β-actin probe. As can be seen in FIG. 19A, TRG3 mRNA (dominant TRG3 mRNA transcript of about 1.7 kb) is HL-60 promyelocytic leukemia cell line, HeLa uterine cancer cell line, chronic myeloid leukemia cell line K-562, lymphocytic leukemia cell line Very high levels were expressed in MOLT-4, Burkitt's lymphoma cell line Raji, colorectal cancer cell line SW480, lung cancer cell line A549 and skin cancer cell line G361.

The upper part of Figure 2b shows the results of Northern blot analysis confirming the expression of TRG4 genes in normal cervical tissues, cervical cancer tissues, cervical cancer metastasis lymph node tissues and cervical cancer cell lines (CaSki and CUMC-6). As can be seen in Figure 2b, the TRG4 gene of the primary cancer genes in the uterine cervical cancer tissue and cervical cancer cancer cell lines CaSki and CUMC-6 cell lines increased gene expression, that is, over 3.0 dominant TRG4 mRNA transcript overexpressed . In Figure 2b, the normal column is the normal cervical cancer tissue, the Cancer column is the cervical cancer tissue, the metastasis column is the cervical cancer metastatic lymph node tissue, and the CaSki column and the CUMC-6 column, respectively. Uterine cancer cells represent the cell line. In the lower part of Fig. 2b, the same sample was hybridized with β-actin probe to confirm the presence of mRNA.

Figure 4a shows the expression of TRG4 primary cancer genes in normal human 12-column multiple tissue tissue (Clontech), brain, heart, rhabdomyo, large intestine, thymus, spleen, kidney, liver, small intestine, placenta, lung and peripheral blood. Northern blot analysis is shown. Figure 4b shows the presence of mRNA by hybridizing the same sample with β-actin probes. As can be seen in Figure 4a, TRG4 mRNA (a dominant TRG4 mRNA transcript of about 3.0 kb) is expressed in normal muscle tissues, but normal brain, heart, large intestine, thymus, spleen, kidney, liver and small intestine. In humans, the placenta, lungs and peripheral blood and leukocytes are very weakly expressed.

Figure 20a shows the results of a Northern blot analysis confirming the expression of TRG4 primary oncogenes in human cancer cell lines, HL-60, HeLa, K-562, MOLT-4, Raji, SW480, A549 and G361 (Clontech). Figure 20b shows the presence of mRNA by hybridizing the same sample with a β-actin probe. As shown in Figure 20a, TRG4 mRNA (dominant TRG4 mRNA transcript of about 3.0 kb) was significantly elevated in HeLa uterine cancer cell line, chronic myeloid leukemia cell line K-562, and colorectal cancer cell line SW480. Expressed.

Figure 2c shows the results of Northern blot analysis confirming the expression of TRG5 primary cancer genes in normal cervical tissue, cervical cancer tissue, cervical cancer metastatic lymph node tissue and cervical cancer cell lines (CaSki and CUMC-6). As can be seen in Figure 2c, the TRG5 proto-oncogene has increased gene expression in cervical cancer tissue and cervical cancer cell lines CaSki and CUMC-6 cell lines, ie overexpressing a dominant TRG5 mRNA transcript of about 1.4 kb. In FIG. 2C, normal columns represent normal cervical tissue, cancer columns represent cervical cancer tissues, metastasis columns represent cervical cancer metastatic lymph node tissues, and CaSki columns and CUMC-6 columns. Each represents a uterine cancer cell line. Figure 2c bottom is to confirm the presence of mRNA by hybridizing the same sample with β-actin probe.

FIG. 5A shows Northern TRG5 prototypic gene expression in normal human 12-row multiple tissue (Clontech), brain, heart, rhabdomyo, colon, thymus, spleen, kidney, liver, small intestine, placenta, lung and peripheral blood leukocyte tissue. The results of the blot analysis are shown. 5B shows the presence of mRNA by hybridizing the same sample with β-actin probe. As can be seen in Figure 5a, TRG5 mRNA (dominant TRG5 mRNA transcript of about 1.4 kb) is expressed in the brain, heart, normal muscle and kidney tissue, but normal colon, thymus, spleen, liver, small intestine , Weakly expressed in placenta, lung and peripheral blood leukocyte tissues.

Figure 21a shows the results of Northern blot analysis confirming the expression of TRG5 primary oncogenes in human cancer cell lines, HL-60, HeLa, K-562, MOLT-4, Raji, SW480, A549 and G361 (Clontech). 21B hybridizes the same sample with β-actin probe to confirm the presence of mRNA. As can be seen in FIG. 21A, TRG5 mRNA (dominant TRG5 mRNA transcript of about 1.4 kb) was expressed in HL-60 promyelocytic leukemia cell line, HeLa uterine cancer cell line, chronic myeloid leukemia cell line K-562, lymphocytic leukemia cell line. Very high levels were expressed in MOLT-4, Burkitt's lymphoma cell line Raji, colorectal cancer cell line SW480, lung cancer cell line A549 and skin cancer cell line G361.

Figure 2d shows the Northern blot analysis confirming the expression of TRG6 primary oncogenes in normal cervical tissue, cervical cancer tissue, cervical cancer metastatic lymph node tissue and cervical cancer cell lines (CaSki and CUMC-6). As can be seen in Figure 2d, the TRG6 proto-oncogene has increased gene expression in cervical cancer tissue and cervical cancer cell lines CaSki and CUMC-6 cell lines, ie overexpressing a dominant TRG6 mRNA transcript of about 7.0 kb. In FIG. 2D, normal fever is normal cervical tissue, cancer fever is cervical cancer tissue, metastasis fever is cervical cancer metastatic lymph node tissue, and CaSki fever and CUMC-6 fever are Each represents a uterine cancer cell line. Figure 2d bottom shows the presence of mRNA by hybridizing the same sample with β-actin probe.

FIG. 6A shows Northern TRG6 prototypic gene expression in normal human 12-row multiple tissue (Clontech), brain, heart, rhabdomyo, colon, thymus, spleen, kidney, liver, small intestine, placenta, lung and peripheral blood leukocyte tissue. The results of the blot analysis are shown. 6B hybridizes the same sample with β-actin probe to confirm the presence of mRNA. As can be seen in FIG. 6A, TRG6 mRNA (dominant TRG6 mRNA transcript of about 7.0 kb) is normal brain, heart, normal muscle, colon, thymus, spleen, kidney, liver, small intestine, placenta, lung and It is not expressed in peripheral blood leukocyte tissues.

22A shows human cancer cell line, HL-60 promyeloid leukemia cell line, HeLa uterine cancer cell line, chronic myeloid leukemia cell line K-562, lymphocyte leukemia cell line MOLT-4, Burkitt's lymphoma cell line Raji, colon cancer cell line SW480, lung cancer The result of Northern blot analysis confirming the expression of TRG6 primary oncogene in cell line A549 and skin cancer cell line G361 (Clontech). Figure 22B shows the presence of mRNA by hybridizing the same sample with β-actin probe. As can be seen in FIG. 22A, TRG6 mRNA (dominant TRG6 mRNA transcript of about 7.0 kb) was expressed at increased levels in HeLa cervical cancer cell line, and skin cancer cell line G361.

Figure 2e shows the Northern blot analysis confirming the expression of TRG7 primary cancer gene in normal cervical tissue, cervical cancer tissue, cervical cancer metastatic lymph node tissue and cervical cancer cell lines (CaSki and CUMC-6). As can be seen in FIG. 2E, the TRG7 proto-oncogene has increased gene expression in cervical cancer tissues and cervical cancer cell lines CaSki and CUMC-6 cell lines, ie over 2.0 dominant TRG7 mRNA transcripts overexpressed. In FIG. 2E, normal fever is normal cervical tissue, cancer fever is cervical cancer tissue, metastasis fever is cervical cancer metastatic lymph node tissue, and CaSki fever and CUMC-6 fever are Each represents a uterine cancer cell line. Figure 2e bottom shows the presence of mRNA by hybridizing the same sample with β-actin probe.

FIG. 7A shows Northern TRG7 prototypic gene expression in normal human 12-row multiple tissue (Clontech), brain, heart, rhabdomyo, colon, thymus, spleen, kidney, liver, small intestine, placenta, lung and peripheral blood leukocyte tissues. The results of the blot analysis are shown. 7B shows the presence of mRNA by hybridizing the same sample with β-actin probe. As can be seen in FIG. 7A, TRG7 mRNA (dominant TRG7 mRNA transcript of about 2.0 kb) is expressed in the heart, normal muscle, kidney and placental tissues and at the same time weakly about 2.4 kb of TRG7 mRNA transcript. It is expressed, but not in normal brain, colon, thymus, spleen, liver, small intestine, lung and peripheral blood leukocyte tissues.

Figure 23a shows the results of Northern blot analysis confirming the expression of TRG7 primary oncogenes in human cancer cell lines, HL-60, HeLa, K-562, MOLT-4, Raji, SW480, A549 and G361 (Clontech). FIG. 23B hybridizes the same sample with β-actin probe to confirm the presence of mRNA. As can be seen in FIG. 23A, TRG7 mRNA (dominant TRG7 mRNA transcript of about 2.0 kb) is HL-60 promyeloid cell leukemia cell line, HeLa uterine cancer cell line, chronic myeloid leukemia cell line K-562, lymphocytic leukemia cell line Very high levels were expressed in MOLT-4, Burkitt's lymphoma cell line Raji, colorectal cancer cell line SW480, lung cancer cell line A549 and skin cancer cell line G361. At the same time, about 2.4 kb of TRG7 mRNA transcript was overexpressed in human cancer cell lines.

Figure 2f shows the results of Northern blot analysis confirming the expression of TRG9 primary cancer genes in normal cervical tissue, cervical cancer tissue, cervical cancer metastatic lymph node tissue and cervical cancer cell lines (CaSki and CUMC-6). As can be seen in FIG. 2F, the TRG9 proto-oncogene has increased gene expression in cervical cancer tissue and cervical cancer cell lines CaSki and CUMC-6 cell lines, ie overexpressing a dominant TRG9 mRNA transcript of about 2.4 kb. In FIG. 2F, normal fever is normal cervical tissue, cancer fever is cervical cancer tissue, metastasis fever is cervical cancer metastatic lymph node tissue, and CaSki fever and CUMC-6 fever are Each represents a uterine cancer cell line. Figure 2f bottom shows the presence of mRNA by hybridizing the same sample with β-actin probe.

FIG. 8A shows Northern TRG9 proteomic gene expression in normal human 12-row multiple tissue (Clontech), brain, heart, rhabdomyo, colon, thymus, spleen, kidney, liver, small intestine, placenta, lung and peripheral blood leukocyte tissues. The results of the blot analysis are shown. 8B shows the presence of mRNA by hybridizing the same sample with β-actin probe. As can be seen in Figure 8a, TRG9 mRNA (dominant TRG3 mRNA transcript of about 2.4 kb) is weakly expressed in the heart, normal muscle, kidney and placental tissue, but normal brain, colon, thymus, spleen, Liver, small intestine, lung and peripheral blood leukocyte tissues are rarely expressed.

Figure 24a shows the results of Northern blot analysis confirming the expression of TRG9 primary oncogenes in human cancer cell lines, HL-60, HeLa, K-562, MOLT-4, Raji, SW480, A549 and G361 (Clontech). FIG. 24B shows the presence of mRNA by hybridizing the same sample with β-actin probe. As can be seen in FIG. 24A, TRG3 mRNA (dominant TRG9 mRNA transcript of about 2.4 kb) was expressed in HL-60 promyelocytic leukemia cell line, HeLa uterine cancer cell line, chronic myeloid leukemia cell line K-562, lymphocytic leukemia cell line. Very high levels were expressed in MOLT-4, Burkitt's lymphoma cell line Raji, colorectal cancer cell line SW480, lung cancer cell line A549 and skin cancer cell line G361. At the same time, expression of about 2.0 kb of TRG9 mRNA transcript was also confirmed.

Figure 2g shows the results of Northern blot analysis confirming the expression of TRG10 primary cancer genes in normal cervical tissue, cervical cancer tissue, cervical cancer metastatic lymph node tissue and cervical cancer cell lines (CaSki and CUMC-6). As can be seen in Figure 2g, the TRG3 proto-oncogene has increased gene expression in cervical cancer tissue and cervical cancer cell lines CaSki and CUMC-6 cell lines, ie overexpressing a dominant TRG10 mRNA transcript of about 6.0 kb. In FIG. 2G, normal fever is normal cervical tissue, cancer fever is cervical cancer tissue, metastasis fever is cervical cancer metastatic lymph node tissue, and CaSki fever and CUMC-6 fever Each represents a uterine cancer cell line. The bottom of Figure 2g hybridizes the same sample with β-actin probe to confirm the presence of mRNA.

9A shows Northern TRG10 proto-oncogenes expressed in normal human 12-row multiple tissue (Clontech), brain, heart, rhabdomyo, colon, thymus, spleen, kidney, liver, small intestine, placenta, lung and peripheral blood leukocyte tissues. The results of the blot analysis are shown. 9B shows the presence of mRNA by hybridizing the same sample with β-actin probe. As can be seen in Figure 9a, TRG10 mRNA (dominant TRG10 mRNA transcript of about 6.0 kb) is expressed in the heart, liver, peripheral white blood cells, and normal spleen tissue, but normal brain, muscle, colon, thymus Expression has not been confirmed in, kidney, small intestine, placenta, and lung tissues.

Figure 25a shows the results of Northern blot analysis confirming the expression of TRG10 primary oncogenes in human cancer cell lines, HL-60, HeLa, K-562, MOLT-4, Raji, SW480, A549 and G361 (Clontech). 25B shows the presence of mRNA by hybridizing the same sample with β-actin probe. As can be seen in FIG. 25A, TRG10 mRNA (dominant TRG10 mRNA transcript of about 6.0 kb) was significantly increased in HeLa cervical cancer cell line, chronic myeloid leukemia cell line K-562, lymphocyte leukemia cell line MOLT-4s. HL-60 promyelocytic leukemia cell line, Burkitt's lymphoma cell line Raji, colon cancer cell line SW480, lung cancer cell line A549 and skin cancer cell line G361. At the same time, about 3.0 kb of TRG10 mRNA transcript was also detected in cancer cell lines.

Figure 2h shows the results of Northern blot analysis confirming the expression of TRG11 primary cancer genes in normal cervical tissue, cervical cancer tissue, cervical cancer metastatic lymph node tissue and cervical cancer cell lines (CaSki and CUMC-6). As can be seen in Figure 2h, the TRG11 proto-oncogene has increased gene expression in cervical cancer tissues, cervical cancer metastatic tissues, cervical cancer cell lines CaSki and CUMC-6 cell lines, that is, dominant TRG11 mRNA of about 1.5 kb. Transcript overexpressed, in FIG. 2H, Normal fever is normal cervical tissue, Cancer fever is cervical cancer tissue, Metastasis fever is cervical cancer metastatic lymph node tissue, and CaSki fever and The CUMC-6 columns represent uterine cancer cell lines, respectively. Figure 2h bottom shows the presence of mRNA by hybridizing the same sample with β-actin probe.

FIG. 10A shows Northern TRG11 prototypic gene expression in normal human 12-row multiple tissue (Clontech), brain, heart, rhabdomyo, colon, thymus, spleen, kidney, liver, small intestine, placenta, lung and peripheral blood leukocyte tissue. The results of the blot analysis are shown. 10B shows the presence of mRNA by hybridizing the same sample with β-actin probe. As can be seen in Figure 10a, TRG11 mRNA (dominant TRG11 mRNA transcript of about 1.5 kb) is expressed in the heart and normal liver tissue, but normal brain, muscle, colon, thymus, spleen, kidney, small intestine Rarely expressed in, placenta, lung and peripheral blood leukocyte tissues.

Figure 26a shows the results of Northern blot analysis confirming the expression of TRG11 primary oncogenes in human cancer cell lines, HL-60, HeLa, K-562, MOLT-4, Raji, SW480, A549 and G361 (Clontech). FIG. 26B hybridizes the same sample with β-actin probe to confirm the presence of mRNA. As can be seen in FIG. 26A, TRG11 mRNA (dominant TRG11 mRNA transcript of about 1.5 kb) was expressed in HL-60 promyelocytic leukemia cell line, HeLa uterine cancer cell line, chronic myeloid leukemia cell line K-562, lymphocytic leukemia cell line. Increased levels in MOLT-4, Burkitt's lymphoma cell line Raji, colorectal cancer cell line SW480, lung cancer cell line A549 and skin cancer cell line G361. In addition, about 1.3 kb of TRG11 mRNA transcript is weakly expressed in cancer cell lines.

Figure 2i shows the results of Northern blot analysis confirming the expression of TRG12 primary cancer genes in normal cervical tissue, cervical cancer tissue, cervical cancer metastatic lymph node tissue and cervical cancer cell lines (CaSki and CUMC-6). As can be seen in Figure 2i, the TRG12 proto-oncogene has increased gene expression in cervical cancer tissues and cervical cancer cell lines CaSki and CUMC-6 cell lines, ie overexpressing a dominant TRG12 mRNA transcript of about 5.0 kb. In FIG. 2I, normal fever is normal cervical tissue, cancer fever is cervical cancer tissue, metastasis fever is cervical cancer metastatic lymph node tissue, and CaSki fever and CUMC-6 fever are Each represents a uterine cancer cell line. Figure 2i bottom shows the presence of mRNA by hybridizing the same sample with β-actin probe.

11A shows Northern TRG12 proto-oncogene expression in normal human 12-row multiple tissue (Clontech), brain, heart, rhabdomyo, colon, thymus, spleen, kidney, liver, small intestine, placenta, lung and peripheral blood leukocyte tissues. The results of the blot analysis are shown. 11b shows the presence of mRNA by hybridizing the same sample with β-actin probe. As can be seen in FIG. 11A, TRG12 mRNA (dominant TRG12 mRNA transcript of about 5.0 kb) is weakly expressed in normal brain, heart, normal muscle, kidney, liver, placenta and peripheral blood leukocyte tissues. There was little expression in normal colon, thymus, spleen, small intestine, and lung tissues.

Figure 27a shows the results of Northern blot analysis confirming the expression of TRG12 primary oncogenes in human cancer cell lines, HL-60, HeLa, K-562, MOLT-4, Raji, SW480, A549 and G361 (Clontech). FIG. 27B hybridizes the same sample with β-actin probe to confirm the presence of mRNA. As can be seen in FIG. 27A, TRG12 mRNA (dominant TRG12 mRNA transcript of about 5.0 kb) was expressed in HL-60 promyelocytic leukemia cell line, HeLa uterine cancer cell line, chronic myeloid leukemia cell line K-562, lymphocytic leukemia cell line. Very high levels were expressed in MOLT-4, Burkitt's lymphoma cell line Raji, colorectal cancer cell line SW480, lung cancer cell line A549 and skin cancer cell line G361.

Figure 2j shows the results of Northern blot analysis confirming the expression of TRG13 primary cancer genes in normal cervical tissue, cervical cancer tissue, cervical cancer metastatic lymph node tissue and cervical cancer cell lines (CaSki and CUMC-6). As can be seen in Figure 2j, the TRG13 proto-oncogene has increased gene expression in cervical cancer tissues and cervical cancer cell lines, CaSki and CUMC-6 cell lines, ie overexpressing a dominant TRG13 mRNA transcript of about 4.0 kb. . In FIG. 2J, normal columns represent normal cervical tissue, cancer rows represent cervical cancer tissues, metastasis columns represent cervical cancer metastatic lymph node tissues, and CaSki columns and CUMC-6 columns, respectively. Uterine cancer cell line. 2J bottom shows the presence of mRNA by hybridizing the same sample with β-actin probe.

12A shows Northern TRG13 proto-oncogenes expressed in normal human 12-row multiple tissue (Clontech), brain, heart, rhabdomyo, colon, thymus, spleen, kidney, liver, small intestine, placenta, lung and peripheral blood leukocyte tissues. The results of the blot analysis are shown. 12B shows the presence of mRNA by hybridizing the same sample with β-actin probe. As can be seen in Figure 12a, TRG13 mRNA (dominant TRG13 mRNA transcript of about 4.0 kb) is expressed in the heart and normal muscle tissue, but normal brain, colon, thymus, spleen, kidney, liver, small intestine , Weakly expressed in placenta, lung and peripheral blood leukocyte tissues. At the same time, about 5.0 kb of TRG13 mRNA transcript was also weakly expressed at the same time.

Figure 28a shows the results of Northern blot analysis confirming the expression of TRG13 primary oncogenes in human cancer cell lines, HL-60, HeLa, K-562, MOLT-4, Raji, SW480, A549 and G361 (Clontech). 28B shows the presence of mRNA by hybridizing the same sample with β-actin probe. As can be seen in FIG. 28A, TRG13 mRNA (dominant TRG13 mRNA transcript of about 4.0 kb) is HL-60 promyelocytic leukemia cell line, HeLa uterine cancer cell line, chronic myeloid leukemia cell line K-562, lymphocytic leukemia cell line Very high levels were expressed in MOLT-4, Burkitt's lymphoma cell line Raji, colorectal cancer cell line SW480, lung cancer cell line A549 and skin cancer cell line G361. At the same time, about 5.0 kb of TRG13 mRNA transcript was also strongly expressed at the same time.

Figure 2k shows the Northern blot analysis results confirming the expression of TRG14 primary cancer genes in normal cervical tissue, cervical cancer tissue, cervical cancer metastatic lymph node tissue and cervical cancer cell lines (CaSki and CUMC-6). As can be seen in Figure 2k, the TRG14 proto-oncogene has increased gene expression in cervical cancer tissue and cervical cancer cell lines CaSki and CUMC-6 cell lines, ie overexpressing a dominant TRG14 mRNA transcript of about 3.5 kb. . In FIG. 2K, normal rows represent normal cervical tissue, cancer rows represent cervical cancer tissues, metastasis rows represent cervical cancer metastatic lymph node tissues, and CaSki rows and CUMC-6 rows, respectively. Uterine cancer cell line. The bottom of Figure 2k hybridizes the same sample with β-actin probe to confirm the presence of mRNA.

FIG. 13A shows Northern TRG14 primary oncogene expression in normal human 12-row multiple tissue (Clontech), brain, heart, rhabdomyo, colon, thymus, spleen, kidney, liver, small intestine, placenta, lung and peripheral blood leukocyte tissues. The results of the blot analysis are shown. FIG. 13B shows the presence of mRNA by hybridizing the same sample with β-actin probe. As can be seen in Figure 13a, TRG14 mRNA (dominant TRG14 mRNA transcript of about 3.5 kb) is expressed in the heart and normal muscle tissue, but normal brain, colon, thymus, spleen, kidney, liver, small intestine , Weakly expressed in placenta, lung and peripheral blood leukocyte tissues.

Figure 29a shows the results of Northern blot analysis confirming the expression of TRG14 primary oncogenes in human cancer cell lines, HL-60, HeLa, K-562, MOLT-4, Raji, SW480, A549 and G361 (Clontech). 29B shows the presence of mRNA by hybridizing the same sample with β-actin probe. As can be seen in FIG. 29A, TRG14 mRNA (dominant TRG14 mRNA transcript of about 3.5 kb) is HL-60 promyelocytic leukemia cell line, HeLa uterine cancer cell line, chronic myeloid leukemia cell line K-562, lymphocytic leukemia cell line Very high levels were expressed in MOLT-4, Burkitt's lymphoma cell line Raji, colorectal cancer cell line SW480, lung cancer cell line A549 and skin cancer cell line G361.

Figure 2 (l) shows the results of Northern blot analysis confirming the expression of TRG15 primary cancer genes in normal cervical tissue, cervical cancer tissue, cervical cancer metastatic lymph node tissue and cervical cancer cell lines (CaSki and CUMC-6). As can be seen in FIG. 2 (l), the TRG15 proto-oncogene has increased gene expression in cervical cancer tissues and cervical cancer cell lines CaSki and CUMC-6 cell lines, ie, a dominant TRG15 mRNA transcript of about 3.5 kb. In Figure 2 (l), normal fever is normal cervical tissue, cancer fever is cervical cancer tissue, metastasis fever is cervical cancer metastatic lymph node tissue, and CaSki fever. And CUMC-6 columns represent uterine cancer cell lines, respectively. 2 (l) at the bottom shows the presence of mRNA by hybridizing the same sample with β-actin probe.

FIG. 14A shows Northern TRG15 proto-oncogene expression in normal human 12-row multiple tissue (Clontech), brain, heart, rhabdomyo, colon, thymus, spleen, kidney, liver, small intestine, placenta, lung and peripheral blood leukocyte tissue The results of the blot analysis are shown. 14B hybridizes the same sample with β-actin probe to confirm the presence of mRNA. As can be seen in Figure 14a, TRG15 mRNA (dominant TRG15 mRNA transcript of about 3.5 kb) is expressed in heart and normal muscle tissue, but normal brain, colon, thymus, spleen, kidney, liver, small intestine Weakly expressed in tissues, placenta, lung and peripheral blood leukocytes. At the same time, about 3.0 kb and 4.0 kb of TRG15 mRNA transcripts are also weakly expressed.

Figure 30a shows the results of Northern blot analysis confirming the expression of TRG15 primary oncogenes in human cancer cell lines, HL-60, HeLa, K-562, MOLT-4, Raji, SW480, A549 and G361 (Clontech). FIG. 30B shows the presence of mRNA by hybridizing the same sample with β-actin probe. As can be seen in FIG. 30A, TRG15 mRNA (dominant TRG15 mRNA transcript of about 3.5 kb) is HL-60 promyelocytic leukemia cell line, HeLa uterine cancer cell line, chronic myeloid leukemia cell line K-562, lymphocytic leukemia cell line Very high levels were expressed in MOLT-4, Burkitt's lymphoma cell line Raji, colorectal cancer cell line SW480, lung cancer cell line A549 and skin cancer cell line G361. At the same time, about 3.0 kb and 4.0 kb of TRG15 mRNA transcripts are also strongly expressed.

Figure 2m shows the results of Northern blot analysis confirming the expression of TRG16 primary cancer genes in normal cervical tissue, cervical cancer tissue, cervical cancer metastatic lymph node tissue and cervical cancer cell lines (CaSki and CUMC-6). As can be seen in Figure 2m, TRG16 proto-oncogenes have increased gene expression in cervical cancer tissues and cervical cancer cell lines CaSki and CUMC-6 cell lines, ie overexpressing a dominant TRG16 mRNA transcript of about 4.5 kb. . In FIG. 2M, normal rows represent normal cervical tissue, cancer rows represent cervical cancer tissues, metastasis rows represent cervical cancer metastatic lymph node tissues, and CaSki rows and CUMC-6 rows, respectively. Uterine cancer cell line. The bottom of FIG. 2m hybridizes the same sample with β-actin probe to confirm the presence of mRNA.

15A shows Northern TRG16 proto-oncogenes expressed in normal human 12-row multiple tissue (Clontech), brain, heart, rhabdomyo, colon, thymus, spleen, kidney, liver, small intestine, placenta, lung and peripheral blood leukocyte tissues. The results of the blot analysis are shown. 15B shows the presence of mRNA by hybridizing the same sample with β-actin probe. As can be seen in Figure 15a, TRG16 mRNA (dominant TRG15 mRNA transcript of about 3.5 kb) is expressed in normal brain and heart tissue, but normal muscle, colon, thymus, spleen, kidney, liver, small intestine Weakly expressed in tissues, placenta, lung and peripheral blood leukocytes. At the same time, about 5.0 kb of TRG16 mRNA transcripts are also very weakly expressed.

Figure 31a shows the results of Northern blot analysis confirming the expression of TRG16 primary oncogenes in human cancer cell lines, HL-60, HeLa, K-562, MOLT-4, Raji, SW480, A549 and G361 (Clontech). FIG. 31B hybridizes the same sample with β-actin probe to confirm the presence of mRNA. As can be seen in FIG. 31A, TRG16 mRNA (dominant TRG16 mRNA transcript of about 4.5 kb) is expressed in HeLa uterine cancer cell line, chronic myeloid leukemia cell line K-562, lymphocyte leukemia cell line MOLT-4, colon cancer cell line SW480, It was expressed at very increased levels in lung cancer cell line A549 and skin cancer cell line G361. At the same time, about 5.0 kb of TRG16 mRNA transcripts are also strongly expressed.

Figure 2n shows the results of Northern blot analysis confirming the expression of TRG17 primary cancer genes in normal cervical tissue, cervical cancer tissue, cervical cancer metastatic lymph node tissue and cervical cancer cell lines (CaSki and CUMC-6). As can be seen in Figure 2n, TRG17 proto-oncogenes have increased gene expression in cervical cancer tissues and cervical cancer cell lines CaSki and CUMC-6 cell lines, ie overexpressing a dominant TRG17 mRNA transcript of about 1.3 kb. . In FIG. 2N, normal rows represent normal cervical tissue, cancer rows represent cervical cancer tissues, metastasis rows represent cervical cancer metastatic lymph node tissues, and CaSki rows and CUMC-6 rows, respectively. Uterine cancer cell line. Figure 2n bottom shows the presence of mRNA by hybridizing the same sample with β-actin probe.

FIG. 16A shows Northern TRG17 proteomic gene expression in normal human 12-row multiple tissue (Clontech), brain, heart, rhabdomyo, colon, thymus, spleen, kidney, liver, small intestine, placenta, lung and peripheral blood leukocyte tissue The results of the blot analysis are shown. 16B shows the presence of mRNA by hybridizing the same sample with β-actin probe. As can be seen in Figure 16a, TRG17 mRNA (dominant TRG17 mRNA transcript of about 1.3 kb) is expressed in normal brain, heart and normal muscle tissue, but normal colon, thymus, spleen, kidney, liver, It is very weakly expressed in small intestine, placenta, lung and peripheral blood leukocyte tissues.

Figure 32a shows the results of Northern blot analysis confirming the expression of TRG17 primary oncogenes in human cancer cell lines, HL-60, HeLa, K-562, MOLT-4, Raji, SW480, A549 and G361 (Clontech). 32B shows the presence of mRNA by hybridizing the same sample with β-actin probe. As can be seen in FIG. 32A, TRG17 mRNA (dominant TRG17 mRNA transcript of about 1.3 kb) was expressed in HL-60 promyelocytic leukemia cell line, HeLa uterine cancer cell line, chronic myeloid leukemia cell line K-562, lymphocytic leukemia cell line. Very high levels were expressed in MOLT-4, Burkitt's lymphoma cell line Raji, colorectal cancer cell line SW480, lung cancer cell line A549 and skin cancer cell line G361.

Figure 2 (o) shows the results of Northern blot analysis confirming the expression of TRG18 primary cancer genes in normal cervical tissue, cervical cancer tissue, cervical cancer metastatic lymph node tissue and cervical cancer cell lines (CaSki and CUMC-6). As can be seen in FIG. 2 (o), the TRG18 proto-oncogene has increased gene expression in cervical cancer tissue and cervical cancer cell lines CaSki and CUMC-6 cell lines, ie, a dominant TRG18 mRNA transcript of about 2.4 kb. This was overexpressed. In FIG. 2 (o), normal fever is normal cervical tissue, cancer fever is cervical cancer tissue, metastasis fever is cervical cancer metastatic lymph node tissue, and CaSki fever and CUMC-6 The rows each represent a uterine cancer cell line. In the lower part of FIG. 2 (o), the same sample was hybridized with β-actin probe to confirm the presence of mRNA.

17A shows Northern TRG18 prototypic gene expression in normal human 12-row multiple tissue (Clontech), brain, heart, rhabdomyo, colon, thymus, spleen, kidney, liver, small intestine, placenta, lung and peripheral blood leukocyte tissues. The results of the blot analysis are shown. 17B shows the presence of mRNA by hybridizing the same sample with β-actin probe. As can be seen in Figure 17a, TRG18 mRNA (dominant TRG18 mRNA transcript of about 2.4 kb) is weakly expressed in normal brain, heart and normal muscle, kidneys, liver and placental tissue, but normal colon, thymus , Spleen, small intestine, lung and peripheral blood leukocyte tissues are not expressed. At the same time, about 1.5 kb of TRG18 mRNA transcripts are also weakly expressed.

Figure 33a shows the results of Northern blot analysis confirming the expression of TRG18 primary oncogenes in human cancer cell lines, HL-60, HeLa, K-562, MOLT-4, Raji, SW480, A549 and G361 (Clontech). 33B hybridizes the same sample with β-actin probe to confirm the presence of mRNA. As can be seen in FIG. 33A, TRG18 mRNA (dominant TRG18 mRNA transcript of about 2.4 kb) was expressed in HL-60 promyelocytic leukemia cell line, HeLa uterine cancer cell line, chronic myeloid leukemia cell line K-562, lymphocytic leukemia cell line. Very high levels were expressed in MOLT-4, Burkitt's lymphoma cell line Raji, colorectal cancer cell line SW480, lung cancer cell line A549 and skin cancer cell line G361. At the same time, about 1.5 kb of TRG18 mRNA transcripts are also strongly expressed.

Figure 2p shows the results of Northern blot analysis confirming the expression of TRG20 primary cancer genes in normal cervical tissue, cervical cancer tissue, cervical cancer metastatic lymph node tissue and cervical cancer cell lines (CaSki and CUMC-6). As can be seen in Figure 2p, gene expression was increased in the cervical cancer tissue and cervical cancer cell lines CaSki and CUMC-6 cell lines, ie, overexpressed the dominant TRG20 mRNA transcript of about 1.3 kb. . In FIG. 2P, normal rows represent normal cervical tissue, cancer rows represent cervical cancer tissues, metastasis rows represent cervical cancer metastatic lymph node tissues, and CaSki rows and CUMC-6 rows, respectively. Uterine cancer cell line. Figure 2p bottom shows the presence of mRNA by hybridizing the same sample with β-actin probe.

18A shows Northern TRG20 proto-oncogenes expressed in normal human 12-row multiple tissue (Clontech), brain, heart, rhabdomyo, colon, thymus, spleen, kidney, liver, small intestine, placenta, lung and peripheral blood leukocyte tissues. The results of the blot analysis are shown. 18B shows the presence of mRNA by hybridizing the same sample with β-actin probe. As can be seen in FIG. 18A, TRG20 mRNA (dominant TRG20 mRNA transcript of about 1.3 kb) is normal brain, heart and normal muscle, colon, thymus, spleen, kidney, liver, small intestine, placenta, lung and Peripheral blood leukocyte tissues are very weakly expressed but not expressed.

Figure 34a shows the results of Northern blot analysis confirming the expression of TRG20 primary cancer genes in human cancer cell lines, HL-60, HeLa, K-562, MOLT-4, Raji, SW480, A549 and G361 (Clontech). 34B shows the presence of mRNA by hybridizing the same sample with β-actin probe. As can be seen in FIG. 34A, TRG20 mRNA (dominant TRG20 mRNA transcript of about 1.3 kb) was expressed in HL-60 promyelocytic leukemia cell line, HeLa uterine cancer cell line, chronic myeloid leukemia cell line K-562, lymphocytic leukemia cell line. Very high levels were expressed in MOLT-4, Burkitt's lymphoma cell line Raji, colorectal cancer cell line SW480, lung cancer cell line A549 and skin cancer cell line G361.

Example 8 Determination of Protein Size after Expression of a TRG Source Cancer Gene in Escherichia Coli

SEQ ID NO: SEQ ID NO: 5; SEQ ID NO: 9; SEQ ID NO: 13; SEQ ID NO: 17; SEQ ID NO: 21; SEQ ID NO: 25; SEQ ID NO: '29; SEQ ID NO: 33; SEQ ID NO: X37; SEQ ID NO: 41; SEQ ID NO: 45; SEQ ID NO: 49; SEQ ID NO: # 53; SEQ ID NO: 57; Alternatively, the entire TRG gene of the original cancer gene of SEQ ID NO: 61 was inserted into the multi-cloning region of the pBAD / thio-Topo vector (Invitrogen, USA), and the generated pBAD / thio-Topo / TRG vector was transferred to E. coli Top10 (Invitrogen, USA). ) Was transfected. In the front of the multi-cloning site of the pBAD / thio-Topo vector, the expression proteins HT-Thioredoxin are inserted. The transfected E. coli was shaken in LB broth, then diluted to 1/100 and incubated for 3 hours. The protein production was induced by adding L-Arabinose (Sigma) of 0.5 mM.

E. coli cells in culture medium before and after the induction of El arabinose were ultrasonically pulverized and then electrophoresed on 12% sodium dodecyl sulfate-polyacrylamide gel (SDS-PAGE).

5A shows the results of SDS-PAGE expression of the protein of the E. coli Top10 strain transfected with the pBAD / thio-Topo / TRG3 vector. A fusion protein band of about 67 kDa was clearly observed after the induction of El arabinose. This 67 kDa fusion protein contains about 15 kDa HT-thioredoxin protein and about 52 kDa TRG3 protein inserted into pBAD / thio-Topo / TRG3 vector.

Figure 5b is a result of confirming the expression pattern of the E. coli Top10 strain of the E. coli strains transfected with the pBAD / thio-Topo / TRG4 vector by SDS-PAGE. This 29 kDa fusion protein contains about 15 kDa HT-thioredoxin protein and about 14 kDa TRG4 protein inserted into the pBAD / thio-Topo / TRG4 vector.

5C shows the results of SDS-PAGE expression of the protein of the E. coli Top10 strain transfected with the pBAD / thio-Topo / TRG5 vector, and a fusion protein band of about 52 kDa was clearly observed after induction of El arabinose. This 52 kDa fusion protein contains about 15 kDa HT-thioredoxin protein and about 37 kDa TRG5 protein inserted into pBAD / thio-Topo / TRG5 vector.

5D shows the results of SDS-PAGE expression of the protein of the E. coli Top10 strain transfected with the pBAD / thio-Topo / TRG6 vector. A fusion protein band of about 24 kDa was clearly observed after the induction of El arabinose. This 24 kDa fusion protein contains about 15 kDa HT-thioredoxin protein and about 9 kDa TRG6 protein inserted into pBAD / thio-Topo / TRG6 vector.

5E shows the results of SDS-PAGE expression of the E. coli Top10 strain transfected with the pBAD / thio-Topo / TRG7 vector. A fusion protein band of about 35 kDa was clearly observed after induction of El arabinose. This 35 kDa fusion protein contains about 15 kDa HT-thioredoxin protein and about 20 kDa TRG7 protein inserted into the pBAD / thio-Topo / TRG7 vector.

5F shows the results of SDS-PAGE expression of the protein of the E. coli Top10 strain transfected with the pBAD / thio-Topo / TRG9 vector. A fusion protein band of about 73 kDa was clearly observed after the induction of El arabinose. This 73 kDa fusion protein contains about 15 kDa HT-thioredoxin protein and about 58 kDa TRG9 protein inserted into pBAD / thio- Topo / TRG9 vector.

Figure 5g is a result of confirming the expression of the protein of E. coli Top10 strain transfected with pBAD / thio-Topo / TRG10 vector by SDS-PAGE, the fusion protein band of about 21 kDa was clearly observed after El arabinose induction. This 21 kDa fusion protein contains about 15 kDa HT-thioredoxin protein and about 6 kDa TRG10 protein inserted into pBAD / thio-Topo / TRG10 vector.

5H shows the results of SDS-PAGE expression of the protein of E. coli Top10 strain transfected with the pBAD / thio-Topo / TRG11 vector, and a fusion protein band of about 22 kDa was clearly observed after El arabinose induction. This 22 kDa fusion protein contains about 15 kDa HT-thioredoxin protein and about 7 kDa TRG11 protein inserted into the pBAD / thio-Topo / TRG11 vector.

5i shows the expression pattern of the E. coli Top10 strain transfected with the pBAD / thio-Topo / TRG12 vector by SDS-PAGE. A fusion protein band of about 29 kDa was clearly observed after El arabinose induction. This 29 kDa fusion protein contains about 15 kDa HT-thioredoxin protein and about 14 kDa TRG12 protein inserted into pBAD / thio-Topo / TRG12 vector.

Figure 5j is a result of confirming the expression of the protein of E. coli Top10 strain transfected with pBAD / thio-Topo / TRG13 vector by SDS-PAGE, a fusion protein band of about 55 kDa was clearly observed after El arabinose induction. This 55 kDa fusion protein contains about 15 kDa HT-thioredoxin protein and about 40 kDa TRG13 protein inserted into the pBAD / thio- Topo / TRG13 vector.

5K shows the results of SDS-PAGE expression of the E. coli Top10 strain transfected with the pBAD / thio-Topo / TRG14 vector. A fusion protein band of about 58 kDa was clearly observed after El arabinose induction. This 58 kDa fusion protein contains about 15 kDa HT-thioredoxin protein and about 43 kDa TRG14 protein inserted into pBAD / thio-Topo / TRG14 vector.

5 (l) shows the results of SDS-PAGE expression of the E. coli Top10 strain transfected with the pBAD / thio-Topo / TRG15 vector, and a fusion protein band of about 57 kDa was clearly observed after induction of El arabinose. . This 57 kDa fusion protein contains about 15 kDa HT-thioredoxin protein and about 42 kDa TRG15 protein inserted into pBAD / thio-Topo / TRG15 vector.

5M shows the results of SDS-PAGE expression of the protein of the E. coli Top10 strain transfected with the pBAD / thio-Topo / TRG16 vector, and a fusion protein band of about 51 kDa was clearly observed after El arabinose induction. This 51 kDa fusion protein contains about 15 kDa HT-thioredoxin protein and about 36 kDa TRG16 protein inserted into pBAD / thio-Topo / TRG16 vector.

5n shows the results of SDS-PAGE expression of the protein of the E. coli Top10 strain transfected with the pBAD / thio-Topo / TRG17 vector. A fusion protein band of about 31 kDa was clearly observed after induction of El arabinose. This 31 kDa fusion protein contains about 15 kDa HT-thioredoxin protein and about 16 kDa TRG17 protein inserted into pBAD / thio- Topo / TRG17 vector.

5 (o) shows the results of SDS-PAGE expression of the E. coli Top10 strain transfected with the pBAD / thio-Topo / TRG18 vector, whereby a fusion protein band of about 57 kDa was clearly observed after induction of El arabinose. . This 57 kDa fusion protein contains about 15 kDa HT-thioredoxin protein and about 42 kDa TRG18 protein inserted into the pBAD / thio-Topo / TRG18 vector.

Figure 5p is a result of confirming the expression of the protein of E. coli Top10 strain transfected with the pBAD / thio-Topo / TRG20 vector by SDS-PAGE, the fusion protein band of about 31 kDa was clearly observed after El arabinose induction. This 31 kDa fusion protein contains about 15 kDa HT-thioredoxin protein and about 16 kDa TRG20 protein inserted into pBAD / thio-Topo / TRG20 vector.

As described above, the primary cancer gene of the present invention is a gene that is involved in human cancer development and at the same time shows a metastasis-inducing ability, and is effective in the diagnosis of cancers including uterine cancer, leukemia, lymphoma, colon cancer, lung cancer and skin cancer. Can be used.

<110> KIM, HYUN-KEE <120> HUMAN PROTOONCOGENE TRG AND PROTEIN ENCODED THEREIN <160> 64 <170> KopatentIn 1.71 <210> 1 <211> 1703 <212> DNA <213> Homo sapiens <400> 1 gggagttgcc ttgacctgca gctccgccac cgcggacccg ccttctgccc tcagcagcag 60 acgctctgtc ccgcccgggc agctctgcga ggcagcggct ggagagggaa ccatggggac 120 tgtgcacgcc cggagtttgg agcctcttcc atcaagtgga cctgattttg gaggattagg 180 agaagaagct gaatttgttg aagttgagcc tgaagctaaa caggaaattc ttgaaaacaa 240 agatgtggtt gttcaacatg ttcattttga tggacttgga aggactaaag atgatatcat 300 catttgtgaa attggagatg ttttcaaggc caaaaaccta attgaggtaa tgcggaaatc 360 tcatgaagcc cgtgaaaaat tgctccgtct tggaattttt agacaagtgg atgttttgat 420 tgacacatgt caaggtgatg acgcacttcc aaatgggtta gacgttacct ttgaagtaac 480 tgaattgagg agattaacgg gcagttataa caccatggtt ggaaacaatg aaggcagtat 540 ggtacttggc ctcaagcttc ctaatcttct tggtcgtgca gaaaaggtga cctttcagtt 600 ttcctatgga acaaaagaaa cttcgtatgg cctgtccttc ttcaaaccac ggcccggaaa 660 cttcgaaaga aatttctctg taaacttata taaagttact ggacagttcc cttggagctc 720 actgcgggag acggacagag gaatgtcagc tgagtacagt tttcccatat ggaagaccag 780 ccacactgtc aagtgggaag gcgtatggcg agaactgggc tgcctctcaa ggacggcgtc 840 atttgctgtt cgaaaagaaa gcggacattc actgaaatca tctctttcgc acgccatggt 900 catcgattct cggaattctt ccatcttacc aaggagaggt gctttgctga aagttaacca 960 ggaactggca ggctacactg gcggggatgt gagcttcatc aaagaagatt ttgaacttca 1020 gttgaacaag caactcatat ttgattcagt tttttcagcg tctttctggg gcggaatgtt 1080 ggtacccatt ggtgataagc cgtcaagcat tgctgatagg ttttaccttg ggggacccac 1140 aagcatccgc ggattcagca tgcacagcat cgggccacag agcgaaggag actacctagg 1200 tggagaagcg tactgggccg gcggcctgca cctctacacc ccattacctt tccggccagg 1260 ccagggtggc tttggagaac ttttccgaac acacttcttt ctcaacgcag gaaacctctg 1320 caacctcaac tatggggagg gccccaaagc tcatattcgt aagctggctg agtgcatccg 1380 ctggtcgtac ggggccggga ttgtcctcag gcttggcaac atcgctcggt tggaacttaa 1440 ttactgcgtc cccatgggag tacagacagg cgacaggata tgtgatggcg tccagtttgg 1500 agctgggata aggttcctgt agccgacacc cctacaggag aagctctggg actggggcag 1560 cagcaaggcg cccatgccac acaccgtctc tcgaggaaac gcggttcagc gattctttga 1620 ctgcggaccc tgtgggaaac cccgtcaata aatgttaaag acacaaaaaa aaaaaaaaaa 1680 aaaaaaaaaa aaaaaaaaaa aaa 1703 <210> 2 <211> 469 <212> PRT <213> Homo sapiens <400> 2 Met Gly Thr Val His Ala Arg Ser Leu Glu Pro Leu Pro Ser Ser Gly   1 5 10 15 Pro Asp Phe Gly Gly Leu Gly Glu Glu Ala Glu Phe Val Glu Val Glu              20 25 30 Pro Glu Ala Lys Gln Glu Ile Leu Glu Asn Lys Asp Val Val Val Gln          35 40 45 His Val His Phe Asp Gly Leu Gly Arg Thr Lys Asp Asp Ile Ile Ile      50 55 60 Cys Glu Ile Gly Asp Val Phe Lys Ala Lys Asn Leu Ile Glu Val Met  65 70 75 80 Arg Lys Ser His Glu Ala Arg Glu Lys Leu Leu Arg Leu Gly Ile Phe                  85 90 95 Arg Gln Val Asp Val Leu Ile Asp Thr Cys Gln Gly Asp Asp Ala Leu             100 105 110 Pro Asn Gly Leu Asp Val Thr Phe Glu Val Thr Glu Leu Arg Arg Leu         115 120 125 Thr Gly Ser Tyr Asn Thr Met Val Gly Asn Asn Glu Gly Ser Met Val     130 135 140 Leu Gly Leu Lys Leu Pro Asn Leu Leu Gly Arg Ala Glu Lys Val Thr 145 150 155 160 Phe Gln Phe Ser Tyr Gly Thr Lys Glu Thr Ser Tyr Gly Leu Ser Phe                 165 170 175 Phe Lys Pro Arg Pro Gly Asn Phe Glu Arg Asn Phe Ser Val Asn Leu             180 185 190 Tyr Lys Val Thr Gly Gln Phe Pro Trp Ser Ser Leu Arg Glu Thr Asp         195 200 205 Arg Gly Met Ser Ala Glu Tyr Ser Phe Pro Ile Trp Lys Thr Ser His     210 215 220 Thr Val Lys Trp Glu Gly Val Trp Arg Glu Leu Gly Cys Leu Ser Arg 225 230 235 240 Thr Ala Ser Phe Ala Val Arg Lys Glu Ser Gly His Ser Leu Lys Ser                 245 250 255 Ser Leu Ser His Ala Met Val Ile Asp Ser Arg Asn Ser Ser Ile Leu             260 265 270 Pro Arg Arg Gly Ala Leu Leu Lys Val Asn Gln Glu Leu Ala Gly Tyr         275 280 285 Thr Gly Gly Asp Val Ser Phe Ile Lys Glu Asp Phe Glu Leu Gln Leu     290 295 300 Asn Lys Gln Leu Ile Phe Asp Ser Val Phe Ser Ala Ser Phe Trp Gly 305 310 315 320 Gly Met Leu Val Pro Ile Gly Asp Lys Pro Ser Ser Ile Ala Asp Arg                 325 330 335 Phe Tyr Leu Gly Gly Pro Thr Ser Ile Arg Gly Phe Ser Met His Ser             340 345 350 Ile Gly Pro Gln Ser Glu Gly Asp Tyr Leu Gly Gly Glu Ala Tyr Trp         355 360 365 Ala Gly Gly Leu His Leu Tyr Thr Pro Leu Pro Phe Arg Pro Gly Gln     370 375 380 Gly Gly Phe Gly Glu Leu Phe Arg Thr His Phe Phe Leu Asn Ala Gly 385 390 395 400 Asn Leu Cys Asn Leu Asn Tyr Gly Glu Gly Pro Lys Ala His Ile Arg                 405 410 415 Lys Leu Ala Glu Cys Ile Arg Trp Ser Tyr Gly Ala Gly Ile Val Leu             420 425 430 Arg Leu Gly Asn Ile Ala Arg Leu Glu Leu Asn Tyr Cys Val Pro Met         435 440 445 Gly Val Gln Thr Gly Asp Arg Ile Cys Asp Gly Val Gln Phe Gly Ala     450 455 460 Gly Ile Arg Phe Leu 465 <210> 3 <211> 16 <212> DNA <213> Artificial Sequence <220> <223> H-T11A Anchor primer for TRG3 <400> 3 aagctttttt ttttta 16 <210> 4 <211> 13 <212> DNA <213> Artificial Sequence <220> <223> H-AP20 Primer for TRG3 <400> 4 aagcttgttg tgc 13 <210> 5 <211> 2576 <212> DNA <213> Homo sapiens <400> 5 ccgaatttcc aacattccat cactaatatt cctcgacgcc accactggga aggttgtgtg 60 caggaacggg ctgctggtga tccgagatga cccagaaggt ctggagttcc cctggggacc 120 gaaacccttc agggaagtca ttgcagggcc cttgcttaga aacaatgggc agtctctgga 180 gagcagcagc ctggaggggt ctcacgtggg cgtctatttc tccgcacatt ggtgtccgcc 240 ctgccgaagc ctcacccggg tcctggtgga atcctaccgg aagatcaagg aggcaggcca 300 gaacttcgag atcatcttcg ttagtgcaga caggtcggag gagtccttca aacagtactt 360 cagtgagatg ccctggctcg ccgtccccta cacggatgag gcccggcggt cgcgcctcaa 420 ccggctgtac ggaatccaag gcatccccac gctcatcatg ctggacccgc agggcgaggt 480 gatcacgcgg caggggcggg tggaggtgct gaacgacgag gactgccggg agttcccctg 540 gcaccccaag cccgtgctgg agctctccga ctccaacgcc gcgcagctta acgagggccc 600 ctgcctcgtc ctttttgtag attctgagga tgacggagag tccgaggcgg ccaagcagct 660 gattcagccg atagctgaga aaatcattgc caagtacaaa gccaaagagg aggaggcacc 720 ccttctgttc ttcgtagccg gggaggatga catgactgac tccctgcgag attacaccaa 780 cctgcctgag gctgcccctt tgctcaccat cctggacatg tcagcccggg ccaagtacgt 840 gatggacgtg gaggagatca cccccgccat cgtggaggcc tttgtgaatg acttcctagc 900 agagaagctc aaaccggagc ccatctagcg tggctccggc ctcctgagac gttatttaaa 960 actcagcctt ctcctcctcc ccctccttcc ttccgccctt ggacttaccc agcgtgcccc 1020 gaatcccacc acccaagtgt ccagcctctc tgtggtgcct tgtttctgca gtaaactcct 1080 cagccagcac cctggggtgt ggaatcagca gcggcagagt ccaccgtgtt tggagactct 1140 gtttgggagc acgggatggc cgggggcccg gccagagcgg ggctgcatgg ctttcgcaaa 1200 gtcactagct tttggtgaag gatctgccag ggtgtcctgg gcagagtgag cgtggagggc 1260 cggtgggtcc cgctcgggct ctgactctga cgtcggcaca cacggccccg gacggccaga 1320 ggggaaccgc cgggtgacac ctgcgtggag gctgagctga gaaagggcct ccgcttagag 1380 ctgcgggtga ggacgtcctt ttctaagcaa cacagtcttc ctcggtctga gagaaaagca 1440 gcccactctt gtgttctcag gcaggggatc tccaaatgca aaaggaagct tgtagaggtt 1500 ttttgggtga gaagaaaaat gtagctaagg taatggttca tatcatacaa acagctccca 1560 cgatcctgaa attctgttaa cgaatccttc tctttggaca tcttcccaag aaacttagcc 1620 ctgagtccta ggaggagcac ctctgcacca gcacggacct ctcgctccac ccagctctgt 1680 gcccaaggcc cctgatctct gctgaggtgc ccacacccac gttcgatcac ccctgccatt 1740 cccttttatt ttcttttttt tgagatggag tcttgctctg tcgcccaggc tggagtgcag 1800 tggcaccatc ttggctcact tcaacctccg cccccaaggt tcaagtgatt ctcctgcctc 1860 agcctcccga gtagctggga ttacaggcac ccgccaccgt acctggctaa tttttgtatt 1920 tttagtagag atggggtttc accatgttgg ctaggctggt ctcgaactcc tgacctcagg 1980 tgatccaccc gcctcggcct cccaaagtgc tgggattaca ggcgtgagcc cccgtgcccg 2040 gccccctttt cttttcaaag cagaactaca aagatgagaa attaccagaa gcctcgcctt 2100 ttcctaagca ccagcggaag gagctgtgcc ccgggatgga gtgagggtgg agggcgcgtc 2160 agccacgggt gggccttgtg tcgccttgta tcggcccagg taggttgttg gcctcttact 2220 tgggctgacc tgacccccga aagagaaaca gacaactctg ttctcaggat tggggatgga 2280 cggcttcggc caagcgtttt agcctcattc actcaggccc cactcagcac tctgccagcc 2340 aagaccattg atttggaaaa tccggtcccc acccgctaat gagctgttga cactgttgtt 2400 ccttgctgaa ttggattgtt gacttgtagt tcagaggcgt acaactagtt ggcgattaga 2460 cttgttatgt gatgttacca gcctgaaatg cgatcacccc gtaggaaata aagcaggcat 2520 ctctggacct caaaaaaaaa aaaaaaaaaa aaaaacaaaa aaaaaaaaaa aaaaaa 2576 <210> 6 <211> 131 <212> PRT <213> Homo sapiens <400> 6 Met Thr Gln Lys Val Trp Ser Ser Pro Gly Asp Arg Asn Pro Ser Gly   1 5 10 15 Lys Ser Leu Gln Gly Pro Cys Leu Glu Thr Met Gly Ser Leu Trp Arg              20 25 30 Ala Ala Ala Trp Arg Gly Leu Thr Trp Ala Ser Ile Ser Pro His Ile          35 40 45 Gly Val Arg Pro Ala Glu Ala Ser Pro Gly Ser Trp Trp Asn Pro Thr      50 55 60 Gly Arg Ser Arg Arg Gln Ala Arg Thr Ser Arg Ser Ser Ser Leu Val  65 70 75 80 Gln Thr Gly Arg Arg Ser Pro Ser Asn Ser Thr Ser Val Arg Cys Pro                  85 90 95 Gly Ser Pro Ser Pro Thr Arg Met Arg Pro Gly Gly Arg Ala Ser Thr             100 105 110 Gly Cys Thr Glu Ser Lys Ala Ser Pro Arg Ser Ser Cys Trp Thr Arg         115 120 125 Arg Ala Arg     130 <210> 7 <211> 16 <212> DNA <213> Artificial Sequence <220> <223> H-T11G anchor primer for TRG4 <400> 7 aagctttttt tttttg 16 <210> 8 <211> 13 <212> DNA <213> Artificial Sequence <220> <223> H-AP9 Primer for TRG4 <400> 8 aagcttcatt ccg 13 <210> 9 <211> 1334 <212> DNA <213> Homo sapiens <400> 9 ggcgccggag gagacgcacg cagctgactt tgtcttctcc gcacgactgt tacagaggtc 60 tccagagcct tctctctcct gtgcaaaatg gcaactctta aggaaaaact cattgcacca 120 gttgcggaag aagaggcaac agttccaaac aataagatca ctgtagtggg tgttggacaa 180 gttggtatgg cgtgtgctat cagcattctg ggaaagtctc tggctgatga acttgctctt 240 gtggatgttt tggaagataa gcttaaagga gaaatgatgg atctgcagca tgggagctta 300 tttcttcaga cacctaaaat tgtggcagat aaagattatt ctgtgaccgc caattctaag 360 attgtagtgg taactgcagg agtccgtcag caagaagggg agagtcggct caatctggtg 420 cagagaaatg ttaatgtctt caaattcatt attcctcaga tcgtcaagta cagtcctgat 480 tgcatcataa ttgtggtttc caacccagtg gacattctta cgtatgttac ctggaaacta 540 agtggattac ccaaacaccg cgtgattgga agtggatgta atctggattc tgctagattt 600 cgctacctta tggctgaaaa acttggcatt catcccagca gctgccatgg atggattttg 660 ggggaacatg gcgactcaag tgtggctgtg tggagtggtg tgaatgtggc aggtgtttct 720 ctccaggaat tgaatccaga aatgggaact gacaatgata gtgaaaattg gaaggaagtg 780 cataagatgg tggttgaaag tgcctatgaa gtcatcaagc taaaaggata taccaactgg 840 gctattggat taagtgtggc tgatcttatt gaatccatgt tgaaaaatct atccaggatt 900 catcccgtgt caacaatggt aaaggggatg tatggcattg agaatgaagt cttcctgagc 960 cttccatgta tcctcaatgc ccggggatta accagcgtta tcaaccagaa gctaaaggat 1020 gatgaggttg ctcagctcaa gaaaagtgca gataccctgt gggacatcca gaaggaccta 1080 aaagacctgt gactagtgag ctctaggctg tagaaattta aaaactacaa tgtgattaac 1140 tcgagccttt agttttcatc catgtacatg gatcacagtt tgctttgatc ttcttcaata 1200 tgtgaatttg ggctcacaga atcaaagcct atgcttggtt taatgcttgc aatctgagct 1260 cttgaacaaa taaaattaac tattgtagtg aaaaaaaaaa aaaaaaaaaa aaaaaagaaa 1320 aaaaaaaaaa aaaa 1334 <210> 10 <211> 334 <212> PRT <213> Homo sapiens <400> 10 Met Ala Thr Leu Lys Glu Lys Leu Ile Ala Pro Val Ala Glu Glu Glu   1 5 10 15 Ala Thr Val Pro Asn Asn Lys Ile Thr Val Val Gly Val Gly Gln Val              20 25 30 Gly Met Ala Cys Ala Ile Ser Ile Leu Gly Lys Ser Leu Ala Asp Glu          35 40 45 Leu Ala Leu Val Asp Val Leu Glu Asp Lys Leu Lys Gly Glu Met Met      50 55 60 Asp Leu Gln His Gly Ser Leu Phe Leu Gln Thr Pro Lys Ile Val Ala  65 70 75 80 Asp Lys Asp Tyr Ser Val Thr Ala Asn Ser Lys Ile Val Val Val Thr                  85 90 95 Ala Gly Val Arg Gln Gln Glu Gly Glu Ser Arg Leu Asn Leu Val Gln             100 105 110 Arg Asn Val Asn Val Phe Lys Phe Ile Ile Pro Gln Ile Val Lys Tyr         115 120 125 Ser Pro Asp Cys Ile Ile Ile Val Val Ser Asn Pro Val Asp Ile Leu     130 135 140 Thr Tyr Val Thr Trp Lys Leu Ser Gly Leu Pro Lys His Arg Val Ile 145 150 155 160 Gly Ser Gly Cys Asn Leu Asp Ser Ala Arg Phe Arg Tyr Leu Met Ala                 165 170 175 Glu Lys Leu Gly Ile His Pro Ser Ser Cys His Gly Trp Ile Leu Gly             180 185 190 Glu His Gly Asp Ser Ser Val Ala Val Trp Ser Gly Val Asn Val Ala         195 200 205 Gly Val Ser Leu Gln Glu Leu Asn Pro Glu Met Gly Thr Asp Asn Asp     210 215 220 Ser Glu Asn Trp Lys Glu Val His Lys Met Val Val Glu Ser Ala Tyr 225 230 235 240 Glu Val Ile Lys Leu Lys Gly Tyr Thr Asn Trp Ala Ile Gly Leu Ser                 245 250 255 Val Ala Asp Leu Ile Glu Ser Met Leu Lys Asn Leu Ser Arg Ile His             260 265 270 Pro Val Ser Thr Met Val Lys Gly Met Tyr Gly Ile Glu Asn Glu Val         275 280 285 Phe Leu Ser Leu Pro Cys Ile Leu Asn Ala Arg Gly Leu Thr Ser Val     290 295 300 Ile Asn Gln Lys Leu Lys Asp Asp Glu Val Ala Gln Leu Lys Lys Ser 305 310 315 320 Ala Asp Thr Leu Trp Asp Ile Gln Lys Asp Leu Lys Asp Leu                 325 330 <210> 11 <211> 16 <212> DNA <213> Artificial Sequence <220> <223> H-T11C anchor primer for TRG5 <400> 11 aagctttttt tttttc 16 <210> 12 <211> 13 <212> DNA <213> Artificial Sequence <220> <223> H-AP11 Primer for TRG5 <400> 12 aagcttcggg taa 13 <210> 13 <211> 3309 <212> DNA <213> Homo sapiens <400> 13 tgggaacggg gagggaagaa ctgagggtct gcagactgga cttttcctgg ctcgacccag 60 gacttgggtt gaggatgcac gggggccacc ttgcccgggg ccactggtgg ctccccagag 120 cctcagaccc acacagccca gaggacgagg ccttcaagcc tgcccctctt ctgctttttt 180 agacagtatt tttagagctg gaaagaaatt ttctagccca actccctgtt tcatgaaaga 240 gaaaagaggc tcagaaaagt ttagagagca gctcagtgtc acactgggag ctgggcacag 300 ccgatctcct tccagaaggg ttctgtctgt atcctttatt ctccgcacat ggaggctgcc 360 ctacctggga aggcacccca gcacctgtga aggacattta ctgctcatcc tcacctgccc 420 cctggccctt gctgccttca ttctgtccca atgccagctc cctggatgtc tgtatgtttg 480 aataccagtt gccatgttag gaaggtcagc tgcacagcca agagtgtaag agtgtaaaga 540 aatgcctttt tttttttttt tgagttggag ttttgctctt gtcgcccagg ctggagtgca 600 gtggcgcgat ctcggctcac tgcaacctct gccccctggg ttcaagcgat tctcctgcct 660 cagcctccct agtagctggg actacagcac ccaccaccac acccagctaa tttttgtatt 720 tttagtagag acaggggttt caccacgttg gccaggctgg tctcgaactc ctgaccttag 780 gtgatccgcc cgcctcagcc tcccaaagtg ttgggattac aggcatgagc cactgcgcct 840 ggccagcaaa tgctttttgt gcagaataca cttctttcag gcattgtcag gtgctgtttt 900 gtttaagctc taactcaccc ctggaataca ggggaatgat gacaaccagc ccagccaggc 960 ctgactcatc atggtcacat ccagccccca cccccggcca actaaccact gcaggctcct 1020 cttccagact caccaggggg cctcgaggcc ccggcatctc ccttggccct gggtgtgggt 1080 tttacaagac tgtgtctttc atgacatcat agcccaacca tgtgagaaga aggagaaggc 1140 ccccctttct tcattaatct gaaaaaaagg aaagtgagaa taggctgatt tttaaaagtt 1200 aaggggcaag cagcattgca ttctggggga acgatcctgg ccacagccgc caaacaaaca 1260 ttcactaggc ctcttctgtt ttcataccct tgtaagtggg ttatgtggtg ggtatggtca 1320 gttttttctt ttttcttttc ttttcttttt tttgagacag agtttcgctt ttgttgcccg 1380 ggctggaatg caatggcgcg attcagctca ctgcaatctc cgcctcccgg gttcaagtga 1440 ttctcctgcc ttagcctcct gaaaagctgg gattacaggg ccctgccacc aagcccagct 1500 aattgtattt ttagtagaga caggatttca ccatgttggc caggccagtc tcaaactcct 1560 gacctcaggt gatccacctg cctcagcctc ccagactgtt gggattacag gcatgagcca 1620 ccacgcctgg ccagtttctt cattttacat atggtcacat tggcgcctag aacagttagg 1680 tcgctcgtca cataggcagt taagtggaga accaggtttc aaaatcaggt aagaaaacca 1740 tcatcattaa ctgagcacca gctgtgctaa gcctgccacg ggcgtatcct tgcagcctca 1800 caacagtggg aggtctgtat cctgaatgtc ctcattttac agatgaggac attgaggaga 1860 agagacttac ccaggctcac acagcagctc agcctgttcc aggcgctggt cagtgcgtgt 1920 tctttgccac cagcctgtca ctccagtggc agctccagaa acggaggctg ttgcttttat 1980 ccctaaactg catccacaga gaagccccaa gaaggaggtt ggggccagct cataaaaagc 2040 ctgaatgcca agccaaggag tggatgcctc cagtcatatt tagaacaaag tcaagtataa 2100 atttacagag aaaaaattct aagacagttg gatgttgtcc tgttggtgag gaagggaaag 2160 gtttttcttg tagggaactg gaaccagccc acaactgcac acttgtgagc tgtcatggaa 2220 acctgatccc caacagcttt tgaggttgtt tgtttgtttg tttgtttacc tgtcttgggc 2280 tttgttgctt ttggcaaaag gtacttcaaa caagggaggg cctggactga gggggaccag 2340 gtcttcttgc tgacctcgtc tacaaaggca aaggaaggca aaggaagctg tctcgggtgt 2400 ttctgaacaa cgtgactcat gaggggcttt ggctacctct tgcgttcccc ctagagatgt 2460 ccaggcctta catttaatcg gctttctctg cggtggggta gagaatggag ctcccgcctt 2520 gcgggcagtg ctaaaggtgg agctggggga ttttcctggg aatgatttga gggctcttga 2580 aagcccatgt gttccaaagc gtctttaact ctgggatagc attggaagcc gctgtcatga 2640 caggacatgg cactggatgg ctggcagaga gccctggctg ggagttaggg agccctgggt 2700 tggaatccag ccccacctct tttatgccac aggtttggtc aagttctctc ccgctcaggg 2760 tagggctgtg aactccctct tacagctaag aacatgcagc ttagtgagga caagaccctt 2820 ctagagcttt acccctaatc cccccccagg agccccgagg ccggcattat tcctccccat 2880 tacaggtgat gagcctcaaa ttcagagagc ttaagcaacc tgctcagggt cacgtctcca 2940 acaggcagta gagtcaaggt ataaaccagg tctgtttttg taccagagtc ccagactaac 3000 tgttggtagg aatcttgtaa ccagtcatgt tttcttcctt gttttggccg ctgggaagct 3060 caaagtcaaa ttcgagaccc ttttttttcc aattgtgctg agtctcctac tagactcgct 3120 tcattctagc tttctgcttt tacctttacc ctaatctttt tatttttatg ctattgtact 3180 ttatttttgt aagttgctga gatatctgtt ttgcaacaag atgggctata tctaaataaa 3240 gacatgatca aaggtttgat ttaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaataaaaa 3300 gaaaaaaaa 3309 <210> 14 <211> 82 <212> PRT <213> Homo sapiens <400> 14 Met Lys Glu Lys Arg Gly Ser Glu Lys Phe Arg Glu Gln Leu Ser Val   1 5 10 15 Thr Leu Gly Ala Gly His Ser Arg Ser Pro Ser Arg Arg Val Leu Ser              20 25 30 Val Ser Phe Ile Leu Arg Thr Trp Arg Leu Pro Tyr Leu Gly Arg His          35 40 45 Pro Ser Thr Cys Glu Gly His Leu Leu Lele Ile Leu Thr Cys Pro Leu      50 55 60 Ala Leu Ala Ala Phe Ile Leu Ser Gln Cys Gln Leu Pro Gly Cys Leu  65 70 75 80 Tyr val         <210> 15 <211> 16 <212> DNA <213> Artificial Sequence <220> <223> H-T11A anchor primer for TRG6 <400> 15 aagctttttt ttttta 16 <210> 16 <211> 13 <212> DNA <213> Artificial Sequence <220> <223> H-AP38 Primer for TRG6 <400> 16 aagcttccag tgc 13 <210> 17 <211> 1778 <212> DNA <213> Homo sapiens <400> 17 ggccattacc aatcgcgaaa ccggctgggc ctctgcgggc gatggggcgg caggccctgc 60 tgcttctcgc gctgtgcgcc acaggcgccc aggggctcta cttccacatc ggcgagaccg 120 agaagcgctg tttcatcgag gaaatccccg acgagaccat ggtcatcggc aactatcgta 180 cccagatgtg ggataagcag aaggaggtct tcctgccctc gacccctggc ctgggcatgc 240 acgtggaagt gaaggacccc gacggcaagg tggtgctgtc ccggcagtac ggctcggagg 300 gccgcttcac gttcacctcc cacacgcccg gtgaccatca aatctgtctg cactccaatt 360 ctaccaggat ggctctcttc gctggtggca aactgcgggt gcatctcgac atccaggttg 420 gggagcatgc caacaactac cctgagattg ctgcaaaaga taagctgacg gagctacagc 480 tccgcgcccg ccagttgctt gatcaggtgg aacagattca gaaggagcag gattaccaaa 540 gggcaagtgc atatctcctt gtaatttgag agggcagttg acctttatac ccactatacc 600 tactcaagtt tctgcttggg agatcagctc tgcagagaat ggaatgagaa gtattggttt 660 agataggttg tttgtttgtt gtttttgaga cggagtttca ctcttgttgc ccatgctgga 720 gtgcaatgcc atgatcttgg ctcactgcaa cctccgcctc cccaggttca agcgattctc 780 ctgcctcagc ctcctgagta gctgggatta caggcatgcg ccaccatgcc tggctaattt 840 tgtactttta gtagagacgg gggtttctcc acgttggtca ggctggtctc gaactcccga 900 catcaagtga tccgcccgcc tcagcctccc aaagtgctgg gattacaggt gtgagctacc 960 gcgccctgcc tgttttgctt ttttatcaaa acattttatt gtggtaaaat ataacaccaa 1020 atgtgtcatt ttaactgtct atatagttca gtggtattaa gtgccttcat aatgttgtgc 1080 taccaacacc atcatccagc tccagaactt tttcatcttc tcaaactaaa aatctgtact 1140 tattttgttt tgtttttgag atggagtctc gctctgttgc ccaggctgga gcgcagtggc 1200 gccatctcgg ctcactgcac cctccgcctc ccaggttcaa gcgattctcc tgcctcagcc 1260 tcccaagtag ctgggattac aggcaagtgc caccatgcgt ggctcatttt tgtgttttta 1320 gtagagactg ggtttcacca tgttggccag gctggtcttg aactcctggc ctcaggcaat 1380 ccactgccgc agcctcccaa agtgttggga ttacaggcgt gagccactgc acccagcaaa 1440 tctgtactta ttataaacaa taacttcccg tttccttttg tcctgacacc caccattcta 1500 ctttctgtct ctatgatcct gactacccta tctcatataa gtggaatcat tcagtatttg 1560 tccttttgtg actggcttat ttcactgagt ataatgttct cacagttcat ccatgttata 1620 gcatgtgtca gaatttctta aggctaatat tccattgtat gcatgtgcca catttcgctt 1680 tcagtagtca tttttaagct ctataaaata aaatgaagaa aggacagttc acaatctaaa 1740 aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaa 1778 <210> 18 <211> 175 <212> PRT <213> Homo sapiens <400> 18 Met Gly Arg Gln Ala Leu Leu Leu Leu Ala Leu Cys Ala Thr Gly Ala   1 5 10 15 Gln Gly Leu Tyr Phe His Ile Gly Glu Thr Glu Lys Arg Cys Phe Ile              20 25 30 Glu Glu Ile Pro Asp Glu Thr Met Val Ile Gly Asn Tyr Arg Thr Gln          35 40 45 Met Trp Asp Lys Gln Lys Glu Val Phe Leu Pro Ser Thr Pro Gly Leu      50 55 60 Gly Met His Val Glu Val Lys Asp Pro Asp Gly Lys Val Val Leu Ser  65 70 75 80 Arg Gln Tyr Gly Ser Glu Gly Arg Phe Thr Phe Thr Ser His Thr Pro                  85 90 95 Gly Asp His Gln Ile Cys Leu His Ser Asn Ser Thr Arg Met Ala Leu             100 105 110 Phe Ala Gly Gly Lys Leu Arg Val His Leu Asp Ile Gln Val Gly Glu         115 120 125 His Ala Asn Asn Tyr Pro Glu Ile Ala Ala Lys Asp Lys Leu Thr Glu     130 135 140 Leu Gln Leu Arg Ala Arg Gln Leu Leu Asp Gln Val Glu Gln Ile Gln 145 150 155 160 Lys Glu Gln Asp Tyr Gln Arg Ala Ser Ala Tyr Leu Leu Val Ile                 165 170 175 <210> 19 <211> 16 <212> DNA <213> Artificial Sequence <220> <223> H-T11G anchor primer for TRG7 <400> 19 aagctttttt tttttg 16 <210> 20 <211> 13 <212> DNA <213> Artificial Sequence <220> <223> H-AP38 Primer for TRG7 <400> 20 aagcttccag tgc 13 <210> 21 <211> 1582 <212> DNA <213> Homo sapiens <400> 21 gttcgggtga gcgaagatgg cggccgagag ggaacctcct ccgctggggg acgggaagcc 60 caccgacttt gaggatctgg aggacggaga ggacctgttc accagcactg tctccaccct 120 agagtcaagt ccatcatctc cagaaccagc tagtcttcct gcagaagata ttagtgcaaa 180 ctccaatggc ccaaaaccca cagaagttgt attagatgat gacagagaag atctttttgc 240 agaagccaca gaagaagttt ctttggacag ccctgcaagg gaacctatcc tatcctcgga 300 accttctcct gcagtcacac ctgtcactcc tactacactc attgctccta gaattgaatc 360 aaagagtatg tctgctcccg tgatctttga tagatccagg gaagagattg aagaagaagc 420 aaatggagac atttttgaca tagaaattgg tgtatcagat ccagaaaaag ttggtgatgg 480 catgaatgcc tatatggcat atagagtaac aacaaagaca tctctttcca tgttcagtaa 540 gagtgaattt tcagtgaaaa gaagattcag cgactttctt ggtttgcaca gcaaattagc 600 aagcaaatat ttacatgttg gttatattgt gccaccagct ccagaaaaga gtatagtagg 660 gatgaccaag gtcaaagtgg gtaaagaaga ctcatcatcc actgagtttg tagaaaaacg 720 gagagcagct cttgaaaggt atcttcaaag aacagtaaaa catccaactt tactacagga 780 tcctgattta aggcagttct tggaaagttc agagctgcct agagcagtta atacacaggc 840 tctgagtggt gcaggaatat tgaggatggt gaacaaggct gccgacgctg tcaacaaaat 900 gacaatcaag atgaatgaat cggatgcatg gtttgaagaa aagcagcagc aatttgagaa 960 tctggatcag caacttagga aacttcatgt cagtgttgaa gccttggtct gtcatagaaa 1020 agaactttca gccaacacag ctgcctttgc taaaagtgct gccatgttag gtaattctga 1080 ggatcatact gctttatcta gagctttgtc tcagcttgca gaggttgagg agaagataga 1140 ccagttacat caagaacaag cttttgctga cttttatatg ttttcagaac tacttagtga 1200 ctacattcgt cttattgctg cagtgaaagg tgtgtttgac catcgaatga agtgctggca 1260 gaaatgggaa gatgctcaaa ttactttgct caaaaaacgt gaagctgaag caaaaatgat 1320 ggttgctaac aaaccagata aaatacagca agctaaaaat gaaataagag agtgggaggc 1380 gaaagtgcaa caaggggaaa gagattttga acagatatct aaaacgattc gaaaagaagt 1440 gggaagattt gagaaagaac gagtgaagga ttttaaaacc gttatcatca agtacttaga 1500 atcactagtt caaacacaac aacagctgat aaaatactgg gaagcattcc tacctgaagc 1560 caaagccatt gcctagcaat aa 1582 <210> 22 <211> 519 <212> PRT <213> Homo sapiens <400> 22 Met Ala Ala Glu Arg Glu Pro Pro Pro Leu Gly Asp Gly Lys Pro Thr   1 5 10 15 Asp Phe Glu Asp Leu Glu Asp Gly Glu Asp Leu Phe Thr Ser Thr Val              20 25 30 Ser Thr Leu Glu Ser Ser Pro Ser Ser Pro Glu Pro Ala Ser Leu Pro          35 40 45 Ala Glu Asp Ile Ser Ala Asn Ser Asn Gly Pro Lys Pro Thr Glu Val      50 55 60 Val Leu Asp Asp Asp Arg Glu Asp Leu Phe Ala Glu Ala Thr Glu Glu  65 70 75 80 Val Ser Leu Asp Ser Pro Ala Arg Glu Pro Ile Leu Ser Ser Glu Pro                  85 90 95 Ser Pro Ala Val Thr Pro Val Thr Pro Thr Thr Leu Ile Ala Pro Arg             100 105 110 Ile Glu Ser Lys Ser Met Ser Ala Pro Val Ile Phe Asp Arg Ser Arg         115 120 125 Glu Glu Ile Glu Glu Glu Ala Asn Gly Asp Ile Phe Asp Ile Glu Ile     130 135 140 Gly Val Ser Asp Pro Glu Lys Val Gly Asp Gly Met Asn Ala Tyr Met 145 150 155 160 Ala Tyr Arg Val Thr Thr Lys Thr Ser Leu Ser Met Phe Ser Lys Ser                 165 170 175 Glu Phe Ser Val Lys Arg Arg Phe Ser Asp Phe Leu Gly Leu His Ser             180 185 190 Lys Leu Ala Ser Lys Tyr Leu His Val Gly Tyr Ile Val Pro Pro Ala         195 200 205 Pro Glu Lys Ser Ile Val Gly Met Thr Lys Val Lys Val Gly Lys Glu     210 215 220 Asp Ser Ser Ser Thr Glu Phe Val Glu Lys Arg Arg Ala Ala Leu Glu 225 230 235 240 Arg Tyr Leu Gln Arg Thr Val Lys His Pro Thr Leu Leu Gln Asp Pro                 245 250 255 Asp Leu Arg Gln Phe Leu Glu Ser Ser Glu Leu Pro Arg Ala Val Asn             260 265 270 Thr Gln Ala Leu Ser Gly Ala Gly Ile Leu Arg Met Val Asn Lys Ala         275 280 285 Ala Asp Ala Val Asn Lys Met Thr Ile Lys Met Asn Glu Ser Asp Ala     290 295 300 Trp Phe Glu Glu Lys Gln Gln Gln Phe Glu Asn Leu Asp Gln Gln Leu 305 310 315 320 Arg Lys Leu His Val Ser Val Glu Ala Leu Val Cys His Arg Lys Glu                 325 330 335 Leu Ser Ala Asn Thr Ala Ala Phe Ala Lys Ser Ala Ala Met Leu Gly             340 345 350 Asn Ser Glu Asp His Thr Ala Leu Ser Arg Ala Leu Ser Gln Leu Ala         355 360 365 Glu Val Glu Glu Lys Ile Asp Gln Leu His Gln Glu Gln Ala Phe Ala     370 375 380 Asp Phe Tyr Met Phe Ser Glu Leu Leu Ser Asp Tyr Ile Arg Leu Ile 385 390 395 400 Ala Ala Val Lys Gly Val Phe Asp His Arg Met Lys Cys Trp Gln Lys                 405 410 415 Trp Glu Asp Ala Gln Ile Thr Leu Leu Lys Lys Arg Glu Ala Glu Ala             420 425 430 Lys Met Met Val Ala Asn Lys Pro Asp Lys Ile Gln Gln Ala Lys Asn         435 440 445 Glu Ile Arg Glu Trp Glu Ala Lys Val Gln Gln Gly Glu Arg Asp Phe     450 455 460 Glu Gln Ile Ser Lys Thr Ile Arg Lys Glu Val Gly Arg Phe Glu Lys 465 470 475 480 Glu Arg Val Lys Asp Phe Lys Thr Val Ile Ile Lys Tyr Leu Glu Ser                 485 490 495 Leu Val Gln Thr Gln Gln Gln Leu Ile Lys Tyr Trp Glu Ala Phe Leu             500 505 510 Pro Glu Ala Lys Ala Ile Ala         515 <210> 23 <211> 16 <212> DNA <213> Artificial Sequence <220> <223> H-T11A anchor primer for TRG9 <400> 23 aagctttttt ttttta 16 <210> 24 <211> 13 <212> DNA <213> Artificial Sequence <220> <223> H-AP9 Primer for TRG9 <400> 24 aagcttcatt ccg 13 <210> 25 <211> 3979 <212> DNA <213> Homo sapiens <400> 25 caaaacttct gcctttaata actttagaaa atttactaat ctatagaact aattgagtag 60 gatataggaa ggatacaagg atataatgtc ctttttataa aagtttagta tagcttcttt 120 acatgtatcc acttgttcca gaaaatgtgc attggttctg aatgtgaaaa tatttaaaga 180 gagaaaggaa cactcaagta agtgtgggct tcagtgggaa ttatcacaaa acattggcaa 240 gtatttttat ttaaattatt ttcaaatttg acttctacag ccaagtggaa ttggtaggct 300 gtagctgtta cactgaaatt tctagtcttt gtaagtgcct cctgaaagtc atttaaaatg 360 gaaaaatatt tcaatgagct tttccttttt tcatatttat ggacatgaat attttattgg 420 agatcattaa ctcctagaat ttgagattat atttccatac aacattttat aaagttatgt 480 tgaacttact acctgttatg tgcaggttat tatgtaacta ttcacagatt gcttcatata 540 ttgctttatc ttcccatcta acttcttaaa gttaaaatcc ggacacacat gttgattatc 600 tagaccagtc attctggaaa ttgtaacact cccacataaa ccccaggaga ctttttcaga 660 atgcaatgtt tctaaatgta ctgttactgg cagtttactc tccagcatat aaggttgcat 720 tttaactttt agattatgaa ctgtgcaaac tttacccaaa actatcttgc atgattccct 780 cctaaatata ttccttgatt aagtaaatct ggcaaatcac tgtttgagct agttacataa 840 aatttgttat caagagaagg cttttctaca agtttccaga ttaacataaa gaaaagaggg 900 aatcacaggg catttaagtg caccttccca ttactttcct taaatcacct catagttagg 960 ctgggcgcgg tggctcacgc ctgtaatccc agcactttgg gaggccgaga cggtggatca 1020 cgaggtcagg ggactgagat catcctggct aacacgatga aacctcatct ctactaaaaa 1080 tacaaataat tagccaggca tggtggcacg cgcctgtagt cccagctact cgggaggcag 1140 agacaggaga atcgtttgca cccgggaggc ggaggttgtt gcagtgagcc gagatcgcgc 1200 caatgcactc cagcctgggc tacagagtga gactccatct caaaaaaaaa caaaaaaaat 1260 cacctcatag tttatgtctg acttactcca aacctcagtt atctgatttg tagtctgtgt 1320 caggaaagtt ttcttcatat ctattctgtc tccctcctct ttgattttaa atttttttct 1380 tttacccagt aggacaaaaa agagcagttg gtcatcatcc ccaatattct tagtcttcag 1440 tatgcttcag gcctctcaat gaacacttaa gtctcaattc ttcagacaaa attgcttaag 1500 ctcttctcct cagtcctatt ttaatgactt tatatttaag aatatagaat tatattttct 1560 tttatattca aattcattac tccagttaag taatagtttg atagtttgat agaatcgaga 1620 gttaagatgt ttctatttga aagtggattc aaccatcaga ccaccagcaa atcggcactt 1680 aatttttgtg ttatctaaca ttttctattg tggaatttta tgattttata ttctcattag 1740 ttataactaa aaagccatgc acacagaatt gtatatcatt ttgccattaa aattttttaa 1800 catattgcag caagcttagt tttatgattg agccacaacc ttttacatat tttttgtatg 1860 aaatattaaa cactaaatgc aagattaact ttcaaaagca aaccctacat taatcaggta 1920 ttatctatgg acatttttgt agaccacttt tgaaatactt attattttgc aacatagact 1980 ggactataca actttcattt aacttttagg tgactgattt aagttgagtg tgcatataga 2040 gaaaaaccta gaaatttatc tcatggcaga tacatttgaa agtacttcag aagaatttat 2100 gctgtatatt aaaactaggc tcaaaataaa tctatcgtat ctttaaaagt ccaattctgt 2160 tattactgtg atgtttgtag tgttactatt aaacattgtg aacatacaca tttttaaaac 2220 aacttgaaac ccattttaaa atctgggtaa gagagaagga atcttcagaa caaaatcaca 2280 tcattagggt gtccagttta tgattgaatt tttaagcaaa ttactgtatt tgaaactaca 2340 acttgatttg gttttcagtt ttaaaaggca acatgtgggt tttatccatt ttatttatac 2400 ctttagattt cagaaacatc ttcatgtttt agatgcattc tacagacatc atgttactta 2460 aaaactcagg gcccctttca tccctttgta cactgaaaaa gttcaattgt tagcaagtaa 2520 gcaattagat ccagttgaat atttaaagtg tttgttgcac agttcattta atgtttcatc 2580 ttatttgact ttttcacata gatataatat cagatttcat taattataaa aagttgccca 2640 gttctgtaat tactgaacag agggaatgac tcaactaatt ggctacatgt tgcaacaaat 2700 ttaggccttt agagttgaag cactgactta aaacgactta cattctgttc tttggtcaaa 2760 tgaccataca tgatatggga caaattgttt cattttgttt gttttttaat aagggaactt 2820 ggtaaagtag ttcctgtcag ataggatttt ctcaagagac aatttaacgt tataaagcct 2880 tctaaaagtg aactaaatat tttataactt tagtaatagc ttggatggtt ttgagaaaat 2940 aacctgtatt tatcacattg tcaaacagaa tttttctttg aatcagacaa gttcaagctc 3000 taaattgatg tgctatatac ttaaaatcct aggaagttat ctgtaaccag tctcttgtct 3060 caggctcttc accttgttac caatcctcgt aagtatgtaa aggaaacata tttttaaaga 3120 agcttaacag taagaaaaaa ttactaaaag atgcaattca aagataggtc ccagtttaac 3180 actgaattgc ttgacttctg tggcttttct ttttctggcc acatttattt atttaagcaa 3240 tttttgtatg ccttgttatt tcatttccat agggattata ttgtatcagt gtttatgtaa 3300 gctggaatca tcctcagttt tttgctgata atttttcaaa taaagataca tggataattg 3360 taaaatacac taactcttag ggtgttgtag tagctgaaac atggagatgc gtagctgtca 3420 tgctttttct gaatggacag gagaaacata agctacggag tattcacttc tgaggatgct 3480 tttccggaaa aagaaaggct agaaaatact cgcacttcct cagaaccctc tttcttgtta 3540 acgggtatct tttgttggtg tgttttgctc ttacattaca gatagactat catatatgac 3600 tttatgaata atttcagtta ttttgctttt gtataagctg tctgaagcct tgctatgctg 3660 tataagttgt gtttgatgga tcagtgtgag tataaaataa agcaaatcac ttttcttttg 3720 tattatctat ggatgccact atgaaagctg acattaagcc actaaagagt tttctatgaa 3780 taagtgtaag taaatgcttt gatatatata aacctaaata aaaagattgt attgatacag 3840 agacattgga gaaggagatt ttaaggcagt tctttaggtt taaaaaggct tgctgtaaaa 3900 tggtgcgtta ttccgtttat taaagatcat attaatgaca aaaaaaaaaa aaaaaaaaaa 3960 aaaaaaaaaa aaaaaaaaa 3979 <210> 26 <211> 56 <212> PRT <213> Homo sapiens <400> 26 Met Val Ala Arg Ala Cys Ser Pro Ser Tyr Ser Gly Gly Arg Asp Arg   1 5 10 15 Arg Ile Val Cys Thr Arg Glu Ala Glu Val Val Ala Val Ser Arg Asp              20 25 30 Arg Ala Asn Ala Leu Gln Pro Gly Leu Gln Ser Glu Thr Pro Ser Gln          35 40 45 Lys Lys Thr Lys Lys Ile Thr Ser      50 55 <210> 27 <211> 16 <212> DNA <213> Artificial Sequence <220> <223> H-T11A anchor primer for TRG10 <400> 27 aagctttttt ttttta 16 <210> 28 <211> 13 <212> DNA <213> Artificial Sequence <220> <223> H-AP9 Primer for TRG10 <400> 28 aagcttcatt ccg 13 <210> 29 <211> 235 <212> DNA <213> Homo sapiens <400> 29 cttccggggt aatgacagtg atgaaatgca ggggacctgg ttgcccccaa gtttcctggc 60 agtgtgtgat actgaggagg tgagcttgtt tctggagctg tgctttaaga ttcatgttac 120 atgtaaagct gtcctcattt gtgactatgg acctatggag ttgggacaat ctctatggga 180 agcagaaggc aaggaccccg gtcattttag gtagaaacaa cagcatgcta atgca 235 <210> 30 <211> 62 <212> PRT <213> Homo sapiens <400> 30 Met Gln Gly Thr Trp Leu Pro Pro Ser Phe Leu Ala Val Cys Asp Thr   1 5 10 15 Glu Glu Val Ser Leu Phe Leu Glu Leu Cys Phe Lys Ile His Val Thr              20 25 30 Cys Lys Ala Val Leu Ile Cys Asp Tyr Gly Pro Met Glu Leu Gly Gln          35 40 45 Ser Leu Trp Glu Ala Glu Gly Lys Asp Pro Gly His Phe Arg      50 55 60 <210> 31 <211> 16 <212> DNA <213> Artificial Sequence <220> <223> H-T11C anchor primer for TRG11 <400> 31 aagctttttt tttttc 16 <210> 32 <211> 13 <212> DNA <213> Artificial Sequence <220> <223> H-AP4 Primer for TRG11 <400> 32 aagcttctca acg 13 <210> 33 <211> 510 <212> DNA <213> Homo sapiens <400> 33 gtagcgcgtc ttgggtctcc cggctgccgc tgctgccgcc gccgcctcgg gtcgtggagc 60 caggagcgac gtcaccgcca tggcaggcat caaagctttg attagtttgt cctttggagg 120 agcaatcgga ctgatgtttt tgatgcttgg atgtgccctt ccaatataca acaaatactg 180 gcccctcttt gttctatttt tttacatcct ttcacctatt ccatactgca tagcaagaag 240 attagtggat gatacagatg ctatgagtaa cgcttgtaag gaacttgcca tctttcttac 300 aacgggcatt gtcgtgtcag cttttggact ccctattgta tttgccagag cacatctgat 360 tgagtgggga gcttgtgcac ttgttctcac aggaaacaca gtcatctttg caactatact 420 aggctttttc ttggtctttg gaagcaatga cgacttcagc tggcagcagt ggtgaaaaga 480 aattactgaa ctattgtcaa atggacttcc 510 <210> 34 <211> 131 <212> PRT <213> Homo sapiens <400> 34 Met Ala Gly Ile Lys Ala Leu Ile Ser Leu Ser Phe Gly Gly Ala Ile   1 5 10 15 Gly Leu Met Phe Leu Met Leu Gly Cys Ala Leu Pro Ile Tyr Asn Lys              20 25 30 Tyr Trp Pro Leu Phe Val Leu Phe Phe Tyr Ile Leu Ser Pro Ile Pro          35 40 45 Tyr Cys Ile Ala Arg Arg Leu Val Asp Asp Thr Asp Ala Met Ser Asn      50 55 60 Ala Cys Lys Glu Leu Ala Ile Phe Leu Thr Thr Gly Ile Val Val Ser  65 70 75 80 Ala Phe Gly Leu Pro Ile Val Phe Ala Arg Ala His Leu Ile Glu Trp                  85 90 95 Gly Ala Cys Ala Leu Val Leu Thr Gly Asn Thr Val Ile Phe Ala Thr             100 105 110 Ile Leu Gly Phe Phe Leu Val Phe Gly Ser Asn Asp Asp Phe Ser Trp         115 120 125 Gln Gln Trp     130 <210> 35 <211> 16 <212> DNA <213> Artificial Sequence <220> <223> H-T11G anchor primer for TRG12 <400> 35 aagctttttt tttttg 16 <210> 36 <211> 13 <212> DNA <213> Artificial Sequence <220> <223> H-AP10 Primer for TRG12 <400> 36 aagcttccac gta 13 <210> 37 <211> 1301 <212> DNA <213> Homo sapiens <400> 37 tcacctccaa gcacagcatg aaggcgcaca tggtcagaca gcacagccgg cgccaagatc 60 tcttacctca gctagaagct ccgagttctc ttactcccag cagtgaactc agcagcccag 120 gccaaagtga gctcactaac atggatcttg ctgcactctt ctctgacaca cctgccaatg 180 ctagtggttc tgcaggtggg tcggatgagg ctctgaactc cggaatcctg actattgacg 240 tcacttctgt gagctcctct ctgggaggga acctccctgc taataatagc tccctagggc 300 cgatggaacc cctggtcctg gtggcccaca gtgatattcc cccaagcctg gacagccctc 360 tggttctcgg gacagcagcc acggttctgc agcagggcag cttcagtgtg gatgacgtgc 420 agactgtgag tgcaggagca ttaggctgtc tggtggctct gcccatgaag aactcgagtg 480 acgacccact ggctttgacc tccaatagta acttagcagc acatatcacc acaccgacct 540 cttcgagcac cccccgagaa aatgccagtg tcccggaact gctggctcca atcaaggtgg 600 agccggactc gccttctcgc ccaggagcag ttgggcagca ggaaggaagc catgggctgc 660 cccagtccac gttgcccagt ccagcagagc agcacggtgc ccaggacaca gagctcagtg 720 caggcactgg caacttctat ttggaaagtg ggggctcagc aagaactgat taccgagcca 780 ttcaactagc caaggaaaaa aagcagagag gagcggggag caatgcagga gcctcacagt 840 ctactcagag aaaaataaaa gaaggcaaaa tgagtcctcc ccatttccat gcaagccaga 900 acagttggtt gtgtgggagc ctcgtggtgc ccagcggagg acggccagga ccagctccag 960 cagctggggt gcagtgcggg gcgcagggcg tccaggtcca gctggtgcag gatgacccct 1020 ccggcgaagg tgtcctgccc tcggcccgcg gcccagccac cttcctcccc ttcctcactg 1080 tggacctgcc cgtctacgtc ctccaggagg tgctcccctc atctggaggc cctgctggac 1140 cggaggccac ccagttccca ggaagcacta tcaacctgca ggatctgcag tgacggcagc 1200 ctcggcctgg gcaggcccaa ggccacggtc taggacacac cttccctgag actcatgaca 1260 tgagcctggg gagacctcat ttggtttccg ttcagaagac c 1301 <210> 38 <211> 391 <212> PRT <213> Homo sapiens <400> 38 Met Lys Ala His Met Val Arg Gln His Ser Arg Arg Gln Asp Leu Leu   1 5 10 15 Pro Gln Leu Glu Ala Pro Ser Ser Leu Thr Pro Ser Ser Glu Leu Ser              20 25 30 Ser Pro Gly Gln Ser Glu Leu Thr Asn Met Asp Leu Ala Ala Leu Phe          35 40 45 Ser Asp Thr Pro Ala Asn Ala Ser Gly Ser Ala Gly Gly Ser Asp Glu      50 55 60 Ala Leu Asn Ser Gly Ile Leu Thr Ile Asp Val Thr Ser Val Ser Ser  65 70 75 80 Ser Leu Gly Gly Asn Leu Pro Ala Asn Asn Ser Ser Leu Gly Pro Met                  85 90 95 Glu Pro Leu Val Leu Val Ala His Ser Asp Ile Pro Pro Ser Leu Asp             100 105 110 Ser Pro Leu Val Leu Gly Thr Ala Ala Thr Val Leu Gln Gln Gly Ser         115 120 125 Phe Ser Val Asp Asp Val Gln Thr Val Ser Ala Gly Ala Leu Gly Cys     130 135 140 Leu Val Ala Leu Pro Met Lys Asn Ser Ser Asp Asp Pro Leu Ala Leu 145 150 155 160 Thr Ser Asn Ser Asn Leu Ala Ala His Ile Thr Thr Pro Thr Ser Ser                 165 170 175 Ser Thr Pro Arg Glu Asn Ala Ser Val Pro Glu Leu Leu Ala Pro Ile             180 185 190 Lys Val Glu Pro Asp Ser Pro Ser Arg Pro Gly Ala Val Gly Gln Gln         195 200 205 Glu Gly Ser His Gly Leu Pro Gln Ser Thr Leu Pro Ser Pro Ala Glu     210 215 220 Gln His Gly Ala Gln Asp Thr Glu Leu Ser Ala Gly Thr Gly Asn Phe 225 230 235 240 Tyr Leu Glu Ser Gly Gly Ser Ala Arg Thr Asp Tyr Arg Ala Ile Gln                 245 250 255 Leu Ala Lys Glu Lys Lys Gln Arg Gly Ala Gly Ser Asn Ala Gly Ala             260 265 270 Ser Gln Ser Thr Gln Arg Lys Ile Lys Glu Gly Lys Met Ser Pro Pro         275 280 285 His Phe His Ala Ser Gln Asn Ser Trp Leu Cys Gly Ser Leu Val Val     290 295 300 Pro Ser Gly Gly Arg Pro Gly Pro Ala Pro Ala Ala Gly Val Gln Cys 305 310 315 320 Gly Ala Gln Gly Val Gln Val Gln Leu Val Gln Asp Asp Pro Ser Gly                 325 330 335 Glu Gly Val Leu Pro Ser Ala Arg Gly Pro Ala Thr Phe Leu Pro Phe             340 345 350 Leu Thr Val Asp Leu Pro Val Tyr Val Leu Gln Glu Val Leu Pro Ser         355 360 365 Ser Gly Gly Pro Ala Gly Pro Glu Ala Thr Gln Phe Pro Gly Ser Thr     370 375 380 Ile Asn Leu Gln Asp Leu Gln 385 390 <210> 39 <211> 16 <212> DNA <213> Artificial Sequence <220> <223> H-T11C anchor primer for TRG13 <400> 39 aagctttttt tttttc 16 <210> 40 <211> 13 <212> DNA <213> Artificial Sequence <220> <223> H-AP11 Primer for TRG13 <400> 40 aagcttcggg taa 13 <210> 41 <211> 1206 <212> DNA <213> Homo sapiens <400> 41 caggtgacag cagggacatg tctcgggaga tgcaggatgt agacctcgct gaggtgaagc 60 ctttggtgga gaaaggggag accatcaccg gcctcctgca agagtttgat gtccaggagc 120 aggacatcga gactttacat ggctctgttc acgtcacgct gtgtgggact cccaagggaa 180 accggcctgt catcctcacc taccatgaca tcggcatgaa ccacaaaacc tgctacaacc 240 cccccttcaa ctacgaggac atgcaggaga tcacccagca ctttgccgtc tgccacgtgg 300 acgcccctgg ccagcaggac ggcgcagcct ccttccccgc agggtacatg tacccctcca 360 tggatcagct ggctgaaatg cttcctggag tccttcaaca gtttgggctg aaaagcatta 420 ttggcatggg aacaggagca ggcgcctaca tcctaactcg atttgctcta aacaaccctg 480 agatggtgga gggccttgtc cttatcaacg tgaacccttg tgcggaaggc tggatggact 540 gggccgcctc caagatctca ggatggaccc aagctctgcc ggacatggtg gtgtcccacc 600 tttttgggaa ggaagaaatg cagagtaacg tggaagtggt ccacacctac cgccagcaca 660 ttgtgaatga catgaacccc ggcaacctgc acctgttcat caatgcctac aacagccggc 720 gcgacctgga gattgagcga ccaatgccgg gaacccacac agtcaccctg cagtgccctg 780 ctctgttggt ggttggggac agctcgcctg cagtggatgc cgtggtggag tgcaactcaa 840 aattggaccc aacaaagacc actctcctca agatggcgga ctgtggcggc ctcccgcaga 900 tctcccagcc ggccaagctc gctgaggcct tcaagtactt cgtgcagggc atgggataca 960 tgccctcggc tagcatgacc cgcctgatgc ggtcccgcac agcctctggt tccagcgtca 1020 cttctctgga tggcacccgc agccgctccc acaccagcga gggcacccga agccgctccc 1080 acaccagcga gggcacccgc agccgctcgc acaccagcga gggggcccac ctggacatca 1140 cccccaactc gggtgctgct gggaacagcg ccgggcccaa gtccatggag gtctcctgct 1200 aggcgg 1206 <210> 42 <211> 394 <212> PRT <213> Homo sapiens <400> 42 Met Ser Arg Glu Met Gln Asp Val Asp Leu Ala Glu Val Lys Pro Leu   1 5 10 15 Val Glu Lys Gly Glu Thr Ile Thr Gly Leu Leu Gln Glu Phe Asp Val              20 25 30 Gln Glu Gln Asp Ile Glu Thr Leu His Gly Ser Val His Val Thr Leu          35 40 45 Cys Gly Thr Pro Lys Gly Asn Arg Pro Val Ile Leu Thr Tyr His Asp      50 55 60 Ile Gly Met Asn His Lys Thr Cys Tyr Asn Pro Pro Phe Asn Tyr Glu  65 70 75 80 Asp Met Gln Glu Ile Thr Gln His Phe Ala Val Cys His Val Asp Ala                  85 90 95 Pro Gly Gln Gln Asp Gly Ala Ala Ser Phe Pro Ala Gly Tyr Met Tyr             100 105 110 Pro Ser Met Asp Gln Leu Ala Glu Met Leu Pro Gly Val Leu Gln Gln         115 120 125 Phe Gly Leu Lys Ser Ile Gle Met Gly Thr Gly Ala Gly Ala Tyr     130 135 140 Ile Leu Thr Arg Phe Ala Leu Asn Asn Pro Glu Met Val Glu Gly Leu 145 150 155 160 Val Leu Ile Asn Val Asn Pro Cys Ala Glu Gly Trp Met Asp Trp Ala                 165 170 175 Ala Ser Lys Ile Ser Gly Trp Thr Gln Ala Leu Pro Asp Met Val Val             180 185 190 Ser His Leu Phe Gly Lys Glu Glu Met Gln Ser Asn Val Glu Val Val         195 200 205 His Thr Tyr Arg Gln His Ile Val Asn Asp Met Asn Pro Gly Asn Leu     210 215 220 His Leu Phe Ile Asn Ala Tyr Asn Ser Arg Arg Asp Leu Glu Ile Glu 225 230 235 240 Arg Pro Met Pro Gly Thr His Thr Val Thr Leu Gln Cys Pro Ala Leu                 245 250 255 Leu Val Val Gly Asp Ser Ser Pro Ala Val Asp Ala Val Val Glu Cys             260 265 270 Asn Ser Lys Leu Asp Pro Thr Lys Thr Thr Leu Leu Lys Met Ala Asp         275 280 285 Cys Gly Gly Leu Pro Gln Ile Ser Gln Pro Ala Lys Leu Ala Glu Ala     290 295 300 Phe Lys Tyr Phe Val Gln Gly Met Gly Tyr Met Pro Ser Ala Ser Met 305 310 315 320 Thr Arg Leu Met Arg Ser Arg Thr Ala Ser Gly Ser Ser Val Thr Ser                 325 330 335 Leu Asp Gly Thr Arg Ser Arg Ser His Thr Ser Glu Gly Thr Arg Ser             340 345 350 Arg Ser His Thr Ser Glu Gly Thr Arg Ser Arg Ser His Thr Ser Glu         355 360 365 Gly Ala His Leu Asp Ile Thr Pro Asn Ser Gly Ala Ala Gly Asn Ser     370 375 380 Ala Gly Pro Lys Ser Met Glu Val Ser Cys 385 390 <210> 43 <211> 16 <212> DNA <213> Artificial Sequence <220> <223> H-T11G anchor primer for TRG14 <400> 43 aagctttttt tttttg 16 <210> 44 <211> 13 <212> DNA <213> Artificial Sequence <220> <223> H-AP20 Primer for TRG14 <400> 44 aagcttgttg tgc 13 <210> 45 <211> 1104 <212> DNA <213> Homo sapiens <400> 45 atggttgcac agaaggtgaa accaaatctt cagactttta ataccattct gaaatgtctc 60 cgaagatttc atgtgtttgc aagatcgcca gccttacagg ttttacgtga aatgaaagcc 120 attggaatag aaccctcgct tgcaacatat caccatatta ttcgcctgtt tgatcaacct 180 ggagaccctt taaagagatc atccttcatc atttatgata taatgaatga atcaatggga 240 aagagatttt ctccaaagga cccggatgat gataagtttt ttcagtcagc catgagcata 300 tgctcatctc tcagagatct agaacttgcc taccaagtac atggcctttt aaaaaccgga 360 gacaactgga aattcattgg acctgatcaa catcgtaatt tctattattc caagttcttc 420 gatttgattt gtctaatgga acaaattgat gttaccttga agtggtatga ggacctgata 480 ccttcagcct actttcccca ctcccaaaca atgatacatc ttctccaagc attggatgtg 540 gccaatcggc tagaagtgat tcctaaaatt tggaaagata gtaaagaata tggtcatact 600 ttccgcagtg acctgagaga agagatcctg atgctcatgg caagggacaa gcacccacca 660 gagcttcagg tggcatttgc tgactgtgct gctgatatca aatctgcgta tgaaagccaa 720 cccatcagac agactgctca ggattggcca gccacctctc tcaactgtat agctatcctc 780 tttttaaggg ctgggagaac tcaggaagcc tggaaaatgt tggggctttt caggaagcat 840 aataagattc ctagaagtga gttgctgaat gagcttatgg acagtgcaaa agtgtctaac 900 agcccttccc aggccattga agtagtagag ctggcaagtg ccttcagctt acctatttgt 960 gagggcctca cccagagagt aatgagtgat tttgcaatca accaggaaca aaaggaagcc 1020 ctaagtaatc taactgcatt gaccagtgac agtgatactg acagcagcag tgacagcgac 1080 agtgacacca gtgaaggcaa atga 1104 <210> 46 <211> 367 <212> PRT <213> Homo sapiens <400> 46 Met Val Ala Gln Lys Val Lys Pro Asn Leu Gln Thr Phe Asn Thr Ile   1 5 10 15 Leu Lys Cys Leu Arg Arg Phe His Val Phe Ala Arg Ser Pro Ala Leu              20 25 30 Gln Val Leu Arg Glu Met Lys Ala Ile Gly Ile Glu Pro Ser Leu Ala          35 40 45 Thr Tyr His His Ile Ile Arg Leu Phe Asp Gln Pro Gly Asp Pro Leu      50 55 60 Lys Arg Ser Ser Phe Ile Ile Tyr Asp Ile Met Asn Glu Ser Met Gly  65 70 75 80 Lys Arg Phe Ser Pro Lys Asp Pro Asp Asp Asp Lys Phe Phe Gln Ser                  85 90 95 Ala Met Ser Ile Cys Ser Ser Leu Arg Asp Leu Glu Leu Ala Tyr Gln             100 105 110 Val His Gly Leu Leu Lys Thr Gly Asp Asn Trp Lys Phe Ile Gly Pro         115 120 125 Asp Gln His Arg Asn Phe Tyr Tyr Ser Lys Phe Phe Asp Leu Ile Cys     130 135 140 Leu Met Glu Gln Ile Asp Val Thr Leu Lys Trp Tyr Glu Asp Leu Ile 145 150 155 160 Pro Ser Ala Tyr Phe Pro His Ser Gln Thr Met Ile His Leu Leu Gln                 165 170 175 Ala Leu Asp Val Ala Asn Arg Leu Glu Val Ile Pro Lys Ile Trp Lys             180 185 190 Asp Ser Lys Glu Tyr Gly His Thr Phe Arg Ser Asp Leu Arg Glu Glu         195 200 205 Ile Leu Met Leu Met Ala Arg Asp Lys His Pro Pro Glu Leu Gln Val     210 215 220 Ala Phe Ala Asp Cys Ala Ala Asp Ile Lys Ser Ala Tyr Glu Ser Gln 225 230 235 240 Pro Ile Arg Gln Thr Ala Gln Asp Trp Pro Ala Thr Ser Leu Asn Cys                 245 250 255 Ile Ala Ile Leu Phe Leu Arg Ala Gly Arg Thr Gln Glu Ala Trp Lys             260 265 270 Met Leu Gly Leu Phe Arg Lys His Asn Lys Ile Pro Arg Ser Glu Leu         275 280 285 Leu Asn Glu Leu Met Asp Ser Ala Lys Val Ser Asn Ser Pro Ser Gln     290 295 300 Ala Ile Glu Val Val Glu Leu Ala Ser Ala Phe Ser Leu Pro Ile Cys 305 310 315 320 Glu Gly Leu Thr Gln Arg Val Met Ser Asp Phe Ala Ile Asn Gln Glu                 325 330 335 Gln Lys Glu Ala Leu Ser Asn Leu Thr Ala Leu Thr Ser Asp Ser Asp             340 345 350 Thr Asp Ser Ser Ser Asp Ser Asp Ser Asp Thr Ser Glu Gly Lys         355 360 365 <210> 47 <211> 16 <212> DNA <213> Artificial Sequence <220> <223> H-T11A anchor primer for TRG15 <400> 47 aagctttttt ttttta 16 <210> 48 <211> 13 <212> DNA <213> Artificial Sequence <220> <223> H-AP15 Primer for TRG15 <400> 48 aagcttacgc aac 13 <210> 49 <211> 1064 <212> DNA <213> Homo sapiens <400> 49 gatcaaaagg ccatgggtag gaggcttgca ggtgtgactt taggatcatg gcttctcccc 60 acgatttttt gggggtcgag ggaagcagag catggtgctc caatcccaga acccagctgc 120 cctcaagctc agaggaccag gtcctggcct ggctgctgct gctgggactc ctccctatgg 180 gcatccctgc atgtcctgtg tgtccctgag gagggacatg gggaactcag gggccacctc 240 tcctcgaact gcggggccag agcagagagc ccttgcacac caccagcctc tcctccctgt 300 gccccaggag ttcatccttt gctgcctggc ccgggaccct gcccgccggc cctctgccca 360 cagcctcctc ttccaccgcg tgctcttcga ggtgcactcg ctgaagctcc tggcagccca 420 ctgcttcatc cagcaccagt acctcatgcc tgagaatgtg gtggaggaga agaccaaggc 480 catggacctg cacgcggtct tggcggagct tccccggccc cgcaggcccc cgctgcagtg 540 gcggtactcg gaagtctcct tcatggagct ggacaaattc ctggaggatg tcaggaatgg 600 aatctaccca ctgatgaact ttgcagccac tcgacccctg gggctgcccc gtgtgctggc 660 cccacccccg gaggaggtcc aaaaggccaa gaccccgacg ccagagccct ttgactctga 720 gaccagaaag gtcatccaga tgcagtgcaa cctggagaga agcgaggaca aggcgcgctg 780 gcatctcact ctgcttctgg tgctggaaga ccggctgcac cggcagctga cctacgacct 840 gctcccaagt aggctgggca ggggacttgg gcggcgggcg gtgaccccag gcggcgggtg 900 ggcagcggcc tcatgcctcg ccttacgcca ttgcctcagc ggacagcgcc caggacctcg 960 cctcggagct cgtgcactat ggcttcctcc acgaggacga ccggatgaag ctggccgcct 1020 tccaggagag caccttcctc aagtaccgtg ggacccaggc ctga 1064 <210> 50 <211> 324 <212> PRT <213> Homo sapiens <400> 50 Met Val Leu Gln Ser Gln Asn Pro Ala Ala Leu Lys Leu Arg Gly Pro   1 5 10 15 Gly Pro Gly Leu Ala Ala Ala Ala Gly Thr Pro Pro Tyr Gly His Pro              20 25 30 Cys Met Ser Cys Val Ser Leu Arg Arg Asp Met Gly Asn Ser Gly Ala          35 40 45 Thr Ser Pro Arg Thr Ala Gly Pro Glu Gln Arg Ala Leu Ala His His      50 55 60 Gln Pro Leu Leu Pro Val Pro Gln Glu Phe Ile Leu Cys Cys Leu Ala  65 70 75 80 Arg Asp Pro Ala Arg Arg Pro Ser Ala His Ser Leu Leu Phe His Arg                  85 90 95 Val Leu Phe Glu Val His Ser Leu Lys Leu Leu Ala Ala His Cys Phe             100 105 110 Ile Gln His Gln Tyr Leu Met Pro Glu Asn Val Val Glu Glu Lys Thr         115 120 125 Lys Ala Met Asp Leu His Ala Val Leu Ala Glu Leu Pro Arg Pro Arg     130 135 140 Arg Pro Pro Leu Gln Trp Arg Tyr Ser Glu Val Ser Phe Met Glu Leu 145 150 155 160 Asp Lys Phe Leu Glu Asp Val Arg Asn Gly Ile Tyr Pro Leu Met Asn                 165 170 175 Phe Ala Ala Thr Arg Pro Leu Gly Leu Pro Arg Val Leu Ala Pro Pro             180 185 190 Pro Glu Glu Val Gln Lys Ala Lys Thr Pro Thr Pro Glu Pro Phe Asp         195 200 205 Ser Glu Thr Arg Lys Val Ile Gln Met Gln Cys Asn Leu Glu Arg Ser     210 215 220 Glu Asp Lys Ala Arg Trp His Leu Thr Leu Leu Leu Val Leu Glu Asp 225 230 235 240 Arg Leu His Arg Gln Leu Thr Tyr Asp Leu Leu Pro Ser Arg Leu Gly                 245 250 255 Arg Gly Leu Gly Arg Arg Ala Val Thr Pro Gly Gly Gly Trp Ala Ala             260 265 270 Ala Ser Cys Leu Ala Leu Arg His Cys Leu Ser Gly Gln Arg Pro Gly         275 280 285 Pro Arg Leu Gly Ala Arg Ala Leu Trp Leu Pro Pro Arg Gly Arg Pro     290 295 300 Asp Glu Ala Gly Arg Leu Pro Gly Glu His Leu Pro Gln Val Pro Trp 305 310 315 320 Asp Pro Gly Leu                 <210> 51 <211> 16 <212> DNA <213> Artificial Sequence <220> <223> H-T11C anchor primer for TRG16 <400> 51 aagctttttt tttttc 16 <210> 52 <211> 13 <212> DNA <213> Artificial Sequence <220> <223> H-AP13 Primer for TRG16 <400> 52 aagcttcggc ata 13 <210> 53 <211> 432 <212> DNA <213> Homo sapiens <400> 53 atggcggaga agtttgacca cctagaggag cacctggaga agttcgtgga gaacattcgg 60 cagctcggca tcatcgtcag tgacttccag cccagcagcc aggccgggct caaccaaaag 120 ctgaatttta ttgttactgg cttacaggat attgacaagt gcagacagca gcttcatgat 180 attactgtac cgttggaagt ttttgaatat atagatcaag gtcgaaatcc ccagctctac 240 accaaagagt gcctggagag ggctctagct aaaaatgagc aagttaaagg caagatcgac 300 accatgaaga aatttaaaag cctgttgatt caagaacttt ctaaagtatt tccggaagac 360 atggctaagt atcgaagcat ccggggggag gatcacccgc cttcttaacc agctcaccct 420 ccctgtgtga ag 432 <210> 54 <211> 135 <212> PRT <213> Homo sapiens <400> 54 Met Ala Glu Lys Phe Asp His Leu Glu Glu His Leu Glu Lys Phe Val   1 5 10 15 Glu Asn Ile Arg Gln Leu Gly Ile Ile Val Ser Asp Phe Gln Pro Ser              20 25 30 Ser Gln Ala Gly Leu Asn Gln Lys Leu Asn Phe Ile Val Thr Gly Leu          35 40 45 Gln Asp Ile Asp Lys Cys Arg Gln Gln Leu His Asp Ile Thr Val Pro      50 55 60 Leu Glu Val Phe Glu Tyr Ile Asp Gln Gly Arg Asn Pro Gln Leu Tyr  65 70 75 80 Thr Lys Glu Cys Leu Glu Arg Ala Leu Ala Lys Asn Glu Gln Val Lys                  85 90 95 Gly Lys Ile Asp Thr Met Lys Lys Phe Lys Ser Leu Leu Ile Gln Glu             100 105 110 Leu Ser Lys Val Phe Pro Glu Asp Met Ala Lys Tyr Arg Ser Ile Arg         115 120 125 Gly Glu Asp His Pro Pro Ser     130 135 <210> 55 <211> 16 <212> DNA <213> Artificial Sequence <220> <223> H-T11G anchor primer for TRG17 <400> 55 aagctttttt tttttg 16 <210> 56 <211> 13 <212> DNA <213> Artificial Sequence <220> <223> H-AP14 Primer for TRG17 <400> 56 aagcttggag ctt 13 <210> 57 <211> 1141 <212> DNA <213> Homo sapiens <400> 57 acacccctag ggcctaaaga tgctgaggtc tgtatggaat tttctgaaac gccacaaaaa 60 gaaatgcatc ttcctgggca cggtccttgg aggagtatat attctgggga aatatggaca 120 gaagaaaatc agagaaatac aggaaaggga ggctgcagaa tacattgccc aagcacgacg 180 acaatatcat tttgaaagta accagaggac ttgcaatatg acagtgctgt ccatgcttcc 240 aacactgaga gaggccttaa tgcagcaact gaattccgag agcctcacag ctctgctaaa 300 aaacaggcct tcaaacaagc tagaaatatg ggaggatctg aagataataa gtttcacaag 360 aagtactgtg gctgtataca gtacctgtat gctggttgtt cttttgcggg tccagttaaa 420 cataattggt ggatatattt acctggataa tgcagcagtt ggcaaaaatg gcactacaat 480 tcttgctccc ccagatgtcc aacagcagta tttatcaagt attcagcacc tacttggaga 540 tggtctgaca gaattgatca ctgtcattaa acaagctgtg cagaaggttt taggaagtgt 600 ttctcttaaa cattctttgt cccttttgga cttggagcaa aaactaaaag aaatcagaaa 660 tctcgttgag cagcataagt cttcttcttg gattaataaa gatggatcca aacctttatt 720 atgccattat atgatgccag atgaagaaac tccattagca gtgcaggcct gtggactttc 780 tcctcgagac attaccacta ttaaacttct caatgaaact agagacatgt tggaaagccc 840 agattttagt acagttttga atacctgttt aaaccgaggt tttagtagac ttctagacaa 900 tatggctgag ttctttcgac ctactgaaca ggacctgcaa catggtaact ctatgaatag 960 tctttccagt gtcagcctgc ctttagctaa gataattcca atagtaaacg gacagatcca 1020 ttcagtttgc agtgaaacac ctagtcattt tgttcaggat ctgttgacaa tggagcaagt 1080 gaaagacttt gctgctaatg tgtatgaagc ttttagtacc cctcagcaac tggagaaatg 1140 a 1141 <210> 58 <211> 373 <212> PRT <213> Homo sapiens <400> 58 Met Leu Arg Ser Val Trp Asn Phe Leu Lys Arg His Lys Lys Lys Cys   1 5 10 15 Ile Phe Leu Gly Thr Val Leu Gly Gly Val Tyr Ile Leu Gly Lys Tyr              20 25 30 Gly Gln Lys Lys Ile Arg Glu Ile Gln Glu Arg Glu Ala Ala Glu Tyr          35 40 45 Ile Ala Gln Ala Arg Arg Gln Tyr His Phe Glu Ser Asn Gln Arg Thr      50 55 60 Cys Asn Met Thr Val Leu Ser Met Leu Pro Thr Leu Arg Glu Ala Leu  65 70 75 80 Met Gln Gln Leu Asn Ser Glu Ser Leu Thr Ala Leu Leu Lys Asn Arg                  85 90 95 Pro Ser Asn Lys Leu Glu Ile Trp Glu Asp Leu Lys Ile Ile Ser Phe             100 105 110 Thr Arg Ser Thr Val Ala Val Tyr Ser Thr Cys Met Leu Val Val Leu         115 120 125 Leu Arg Val Gln Leu Asn Ile Ile Gly Gly Tyr Ile Tyr Leu Asp Asn     130 135 140 Ala Ala Val Gly Lys Asn Gly Thr Thr Ile Leu Ala Pro Pro Asp Val 145 150 155 160 Gln Gln Gln Tyr Leu Ser Ser Ile Gln His Leu Leu Gly Asp Gly Leu                 165 170 175 Thr Glu Leu Ile Thr Val Ile Lys Gln Ala Val Gln Lys Val Leu Gly             180 185 190 Ser Val Ser Leu Lys His Ser Leu Ser Leu Leu Asp Leu Glu Gln Lys         195 200 205 Leu Lys Glu Ile Arg Asn Leu Val Glu Gln His Lys Ser Ser Ser Trp     210 215 220 Ile Asn Lys Asp Gly Ser Lys Pro Leu Leu Cys His Tyr Met Met Pro 225 230 235 240 Asp Glu Glu Thr Pro Leu Ala Val Gln Ala Cys Gly Leu Ser Pro Arg                 245 250 255 Asp Ile Thr Thr Ile Lys Leu Leu Asn Glu Thr Arg Asp Met Leu Glu             260 265 270 Ser Pro Asp Phe Ser Thr Val Leu Asn Thr Cys Leu Asn Arg Gly Phe         275 280 285 Ser Arg Leu Leu Asp Asn Met Ala Glu Phe Phe Arg Pro Thr Glu Gln     290 295 300 Asp Leu Gln His Gly Asn Ser Met Asn Ser Leu Ser Ser Val Ser Leu 305 310 315 320 Pro Leu Ala Lys Ile Ile Pro Ile Val Asn Gly Gln Ile His Ser Val                 325 330 335 Cys Ser Glu Thr Pro Ser His Phe Val Gln Asp Leu Leu Thr Met Glu             340 345 350 Gln Val Lys Asp Phe Ala Ala Asn Val Tyr Glu Ala Phe Ser Thr Pro         355 360 365 Gln Gln Leu Glu Lys     370 <210> 59 <211> 16 <212> DNA <213> Artificial Sequence <220> <223> H-T11C anchor primer for TRG18 <400> 59 aagctttttt tttttc 16 <210> 60 <211> 13 <212> DNA <213> Artificial Sequence <220> <223> H-AP18 Primer for TRG18 <400> 60 aagcttagag gca 13 <210> 61 <211> 449 <212> DNA <213> Homo sapiens <400> 61 ctgctggttg aggctgtgtg ggtgggggac gggccgaggc gatggcggag aagtttgacc 60 acctagagga gcacctggag aagttcgtgg agaacattcg gcagctcggc atcatcgtca 120 gtgacttcca gcccagcagc caggccgggc tcaaccaaaa gctgaatttt attgttactg 180 gcttacagga tattgacaag tgcagacagc agcttcatga tattactgta ccgttagaag 240 tttttgaata tatagatcaa ggtcgaaatc cccagctcta caccaaagag tgcctggaga 300 gggctctagc taaaaatgag caagttaaag gcaagatcga caccatgaag aaatttaaaa 360 gcctgttgat tcaagaactt tctaaagtat ttccggaaga catggctaag tatcgaagca 420 tccgggggga ggatcacccg ccttcttaa 449 <210> 62 <211> 135 <212> PRT <213> Homo sapiens <400> 62 Met Ala Glu Lys Phe Asp His Leu Glu Glu His Leu Glu Lys Phe Val   1 5 10 15 Glu Asn Ile Arg Gln Leu Gly Ile Ile Val Ser Asp Phe Gln Pro Ser              20 25 30 Ser Gln Ala Gly Leu Asn Gln Lys Leu Asn Phe Ile Val Thr Gly Leu          35 40 45 Gln Asp Ile Asp Lys Cys Arg Gln Gln Leu His Asp Ile Thr Val Pro      50 55 60 Leu Glu Val Phe Glu Tyr Ile Asp Gln Gly Arg Asn Pro Gln Leu Tyr  65 70 75 80 Thr Lys Glu Cys Leu Glu Arg Ala Leu Ala Lys Asn Glu Gln Val Lys                  85 90 95 Gly Lys Ile Asp Thr Met Lys Lys Phe Lys Ser Leu Leu Ile Gln Glu             100 105 110 Leu Ser Lys Val Phe Pro Glu Asp Met Ala Lys Tyr Arg Ser Ile Arg         115 120 125 Gly Glu Asp His Pro Pro Ser     130 135 <210> 63 <211> 16 <212> DNA <213> Artificial Sequence <220> <223> H-T11A anchor primer for TRG20 <400> 63 aagctttttt ttttta 16 <210> 64 <211> 13 <212> DNA <213> Artificial Sequence <220> <223> H-AP13 Primer for TRG20 <400> 64 aagcttcggc ata 13  

Claims (7)

Human protoprotein having the amino acid sequence of SEQ ID NO: 6. Human primary cancer gene encoding the primary cancer protein of claim 1. The method of claim 2, Said gene has a nucleotide sequence corresponding to base numbers 87 to 482 of SEQ ID NO: 5. The human primary cancer gene according to claim 2, wherein the gene has a nucleotide sequence of SEQ ID NO: 5. A vector comprising the proto-oncogene of any one of claims 2 to 4. Claim 1 cancer diagnostic kit comprising the original cancer protein. A cancer diagnostic kit comprising the original oncogene of any one of claims 2 to 4.

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