WO2006109941A1

WO2006109941A1 - Human cancer suppressor gene, protein encoded therein

Info

Publication number: WO2006109941A1
Application number: PCT/KR2006/001174
Authority: WO
Inventors: Hyun-Kee Kim; Jin-Woo Kim
Original assignee: Hyun-Kee Kim; Jin-Woo Kim
Priority date: 2005-03-30
Filing date: 2006-03-30
Publication date: 2006-10-19
Also published as: KR100689276B1; JP2008534006A; CN101184774A; EP1866333A4; EP1866333A1; CA2602976A1; KR20060104265A

Abstract

Disclosed are a human cancer suppressor gene, a protein encoded therein, an expression vector containing the same and a microorganism transformed with the vector. The cancer suppressor gene of the present invention may be effectively used for diagnosing, preventing and treating human cancers.

Description

HUMAN CANCER SUPPRESSOR GENE, PROTEIN ENCODED THEREIN

TECHNICAL FIELD

The present invention relates to a human cancer suppressor gene, a protein

encoded therein, an expression vector containing the same and a cell transformed with

the vector.

BACKGROUND ART

Tumor suppressor gene products function to suppress normal cells from being transformed into certain cancer cells, and therefore loss of this function of the tumor

suppressor gene products allows the normal cells to become malignant transformants

(Klein, G., FASEB J, 7, 821-825 (1993)). In order to allow cancer cells to grow into a

cancer, the cells should lose a function to control the normal copy number of a tumor

suppressor gene. It was found that modification in a coding sequence of a p53 tumor

suppressor gene is one of the most general genetic changes in the human cancers (Bishop, J.M., Cell, 64, 235-248 (1991); and Weinberg, R.A., Science, 254, 1138-1146

(1991)).

However, it was estimated that only some of breast cancer tissues exhibit a p53

mutation because the p53 mutation reported in the breast cancer amounts to a range of 30 % of the total breast cancer (Keen, J.C. & Davidson, N. E., Cancer, 97, 825-833 (2003)) and Borresen-Dale, A-L., Human Mutation, 21, 292-300 (2003)). Also, it was estimated that only some of leukemia tissues exhibit a p53 mutation because the p53

mutation reported in the leukemia amounts to a range of 20 % of the total leukemia (Boyapati, A., et al., Acta Haematol., 111(1-2), 100-106 (2004)).

The p53 mutation accounts to at least 50 % of the liver cancer especially in the

region exposed to aflatoxin Bl or having a high frequency of infection by hepatitis B virus, and it is mainly characterized by a missense mutation at a codon 249 in the p53

tumor gene (Montesano, R. et al, J. Natl. Cancer Inst., 89, 1844-1851 (1997);

Szymanska, K. & Hainaut, P. Acta Biochimica Polonica, 50, 231-238 (2003)).

However, it was reported that the p53 mutation amounts to nothing but a range of 30 %

of the liver cancer in U.S. and Western Europe, and there is no hot spot in which such

mutation occurs frequently (Szymanska, K. & Hainaut, P. Acta Biochimica Polonica, 50, 231-238 (2003)).

Meanwhile, it was estimated that only some of the cervical cancer tissues exhibit

a p53 mutation because the p53 mutation reported in the cervical cancer amounts to only a range of 2 to 11 % of the toatal cervical cancer (Crook, T. et al., Lancet, 339,

1070-1073 (1992); and Busby-Earle, R.M.C. et al., BrJ. Cancer, 69, 732-737 (1994)). Also, it was reported that the mutation frequency of a p53 tumor suppressor gene

in the non-small-cell lung cancer and the small-cell lung cancer amounts to

approximately 50 % and 70 % of the total lung cancer, respectively (Takahashi, T. et ah, Science, 246, 491-494 1989; Bodner, S.M. et al., Oncogene, 7, 743-749 (1992); Mao, L. Lung Cancer, 34, S27-S34 (2001)). Smoking is one of the most critical factors in

development and progress of the lung cancer, and other various tumor suppressor genes and cancer genes in addition to the p53 are concurrently involved in the development and the progress of the lung cancer (Osada, H. & Takahashi, T. Oncogene, 21,

7421-7434 (2002)). Accordingly, the present inventors have ardently attempted to separate a novel tumor suppressor gene from normal tissues such as breast, liver, cervix, lungs, etc. using

an mRNA differential display (DD) method for more effectively displaying genes

differentially expressed between the normal tissues such as the breast, the liver, the

cervix and the lungs and the cancer tissues such as the breast cancer, the liver cancer,

the cervical cancer and the lung cancer (Liang, P. and Pardee, A. B., Science, 257, 967-971 (1992); and Liang, P. et al., Cancer Res., 52, 6966-6968 (1993)).

DISCLOSURE OF INVENTION

Accordingly, the present invention is designed to solve the problems of the prior

art, and therefore it is an object of the present invention to provide a novel human

cancer suppressor gene.

It is another object of the present invention to provide a cancer suppressor protein encoded by the cancer suppressor gene. It is still another object of the present invention to provide an expression vector

including the cancer suppressor gene.

In order to accomplish one of the above objects, the present invention provides a human cancer suppressor gene having a DNA sequence selected from the group

consisting of 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; SEQ ID NO: 61; SEQ ID NO: 65; SEQ ID NO: 69; SEQ ID NO: 73; SEQ ID NO: 77; SEQ ID NO: 81; SEQ ID NO: 85; SEQ ID NO: 89; SEQ ID NO: 93; SEQ ID

NO: 97; SEQ ID NO: 101; SEQ ID NO: 105; SEQ ID NO: 109 and SEQ ID NO: 113.

According to another of the above objects, the present invention provides a

human cancer suppressor protein having an amino acid sequence selected from the

group consisting of SEQ ID NO: 2; 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; SEQ ID NO: 62; SEQ ID NO: 66; SEQ ID NO: 70; SEQ ID NO: 74;

SEQ ID NO: 78; SEQ ID NO: 82; SEQ ID NO: 86; SEQ ID NO: 90; SEQ ID NO: 94; SEQ ID NO: 98; SEQ ID NO: 102; SEQ ID NO: 106; SEQ ID NO: 110; and SEQ ID

NO: 114.

According to still another of the above objects, the present invention provides an

expression vector including the cancer suppressor gene.

According to yet another of the above objects, the present invention provides a

cell transformed with the expression vector.

BRIEF DESCRIPTION OF THE DRAWINGS

These and other features, aspects, and advantages of preferred embodiments of the present invention will be more fully described in the following detailed description,

taken accompanying drawings. In the drawings:

FIG. 1 is a gel diagram showing a PCR result using a random 5'-13-mer primer

H-AP33 of SEQ ID NO: 3 and an anchored oligo-dT primer of SEQ ID NO: 4;

FIG. 2 is a gel diagram showing a PCR result using a random 5'-13-mer primer H-APlO of SEQ ID NO: 7 and an anchored oligo-dT primer of SEQ ID NO: 8;

FIG. 3 is a gel diagram showing a PCR result using a random 5'-13-mer primer

H-AP5 of SEQ ID NO: 11 and an anchored oligo-dT primer of SEQ ID NO: 12;

FIG. 4 is a gel diagram showing a PCR result using a random 5'-13-mer primer

H-AP2 of SEQ ID NO: 15 and an anchored oligo-dT primer of SEQ ID NO: 16;

FIG. 5 is a gel diagram showing a PCR result using a random 5'-13-mer primer

H-AP8 of SEQ ID NO: 19 and an anchored oligo-dT primer of SEQ ID NO: 20;

FIG. 6 is a gel diagram showing a PCR result using a random 5'-13-mer primer

H-AP7 of SEQ ID NO: 23 and an anchored oligo-dT primer of SEQ ID NO: 24;

FIG. 7 is a gel diagram showing a PCR result using a random 5'-13-mer primer

H-APl 1 of SEQ ID NO: 27 and an anchored oligo-dT primer of SEQ ID NO: 28;

FIG. 8 is a gel diagram showing a PCR result using a random 5'-13-mer primer H-AP3 of SEQ ID NO: 31 and an anchored oligo-dT primer of SEQ ID NO: 32;

FIG. 9 is a gel diagram showing a PCR result using a random 5'-13-mer primer

H-AP4 of SEQ ID NO: 35 and an anchored oligo-dT primer of SEQ ID NO: 36;

FIG. 10 is a gel diagram showing a PCR result using a random 5'-13-mer primer

H-AP8 of SEQ ID NO: 39 and an anchored oligo-dT primer of SEQ ID NO: 40;

FIG. 11 is a gel diagram showing a PCR result using a random 5'-13-mer primer

H-AP33 of SEQ ID NO: 43 and an anchored oligo-dT primer of SEQ ID NO: 44; FIG. 12 is a gel diagram showing a PCR result using a random 5'-13-mer primer

H-AP35 of SEQ ID NO: 47 and an anchored oligo-dT primer of SEQ ID NO: 48;

FIG. 13 is a gel diagram showing a PCR result using a random 5'-13-mer primer

H-AP3 of SEQ ID NO: 51 and an anchored oligo-dT primer of SEQ ID NO: 52; FIG. 14 is a gel diagram showing a PCR result using a random 5'-13-mer primer H-AP12 of SEQ ID NO: 55 and an anchored oligo-dT primer of SEQ ID NO: 56;

FIG. 15 is a gel diagram showing a PCR result using a random 5'-13-mer primer

H-AP12 of SEQ ID NO: 59 and an anchored oligo-dT primer of SEQ ID NO: 60;

FIG. 16 is a gel diagram showing a PCR result using a random 5'-13-mer primer

H-AP7 of SEQ ID NO: 63 and an anchored oligo-dT primer of SEQ ID NO: 64;

FIG. 17 is a gel diagram showing a PCR result using a random 5'-13-mer primer

H-AP8 of SEQ ID NO: 67 and an anchored oligo-dT primer of SEQ ID NO: 68;

FIG. 18 is a gel diagram showing a PCR result using a random 5'-13-mer primer

H-APlO of SEQ ID NO: 71 and an anchored oligo-dT primer of SEQ ID NO: 72;

FIG. 19 is a gel diagram showing a PCR result using a random 5'-13-mer primer

H-AP16 of SEQ ID NO: 75 and an anchored oligo-dT primer of SEQ ID NO: 76;

FIG. 20 is a gel diagram showing a PCR result using a random 5'-13-mer primer

H-AP2 of SEQ ID NO: 79 and an anchored oligo-dT primer of SEQ ID NO: 80; FIG. 21 is a gel diagram showing a PCR result using a random 5'-13-mer primer

H-AP9 of SEQ ID NO: 83 and an anchored oligo-dT primer of SEQ ID NO: 84;

FIG. 22 is a gel diagram showing a PCR result using a random 5'-13-mer primer H-AP9 of SEQ ID NO: 87 and an anchored oligo-dT primer of SEQ ID NO: 88;

FIG. 23 is a gel diagram showing a PCR result using a random 5'-13-mer primer

H-AP32 of SEQ ID NO: 91 and an anchored oligo-dT primer of SEQ ID NO: 92;

FIG. 24 is a gel diagram showing a PCR result using a random 5'-13-mer primer

H-APlO of SEQ ID NO: 95 and an anchored oligo-dT primer of SEQ ID NO: 96;

FIG. 25 is a gel diagram showing a PCR result using a random 5'-13-mer primer H-AP22 of SEQ ID NO: 99 and an anchored oligo-dT primer of SEQ ID NO: 100;

FIG. 26 is a gel diagram showing a PCR result using a random 5'-13-mer primer

H-AP 12 of SEQ ID NO: 103 and an anchored oligo-dT primer of SEQ ID NO: 104;

FIG. 27 is a gel diagram showing a PCR result using a random 5'-13-mer primer

H-APlO of SEQ ID NO: 107 and an anchored oligo-dT primer of SEQ ID NO: 108;

FIG. 28 is a gel diagram showing a PCR result using a random 5'-13-mer primer

H-AP12 of SEQ ID NO: 111 and an anchored oligo-dT primer of SEQ ID NO: 112; and

FIG. 29 is a gel diagram showing a PCR result using a random 5'-13-mer primer

H-AP4 of SEQ ID NO: 115 and an anchored oligo-dT primer of SEQ ID NO: 116.

FIGs. 30 to FIG. 58 are diagrams showing SDS-PAGE analysis results of a GIG8 gene product (FIG. 30); a GIGlO gene product (FIG. 31); a GIGl 3 gene product (FIG.

32); a GIGl 5 gene product (FIG. 33); a GIGl 6 gene product (FIG. 34); a GIG24 gene

product (FIG. 35); a GIG26 gene product (FIG. 36); a GIG29 gene product (FIG. 37); a

GIG30 gene product (FIG. 38); a GIG32 gene product (FIG. 39); a GIG33 gene product

(FIG. 40); a GIG34 gene product (FIG. 41); a GIG35 gene product (FIG. 42); a GIG38 gene product (FIG. 43); a GIG39 gene product (FIG. 44); a GIG40 gene product (FIG.

45); a GIG42 gene product (FIG. 46); a GIG43 gene product (FIG. 47); a GIG46 gene

product (FIG. 48); a PIG33 gene product (FIG. 49); a PIG35 gene product (FIG. 50); a

PIG36 gene product (FIG. 51); an MIG20 gene product (FIG. 52); a PIG49 gene product (FIG. 53); a PIG51 gene product (FIG. 54); an MIGl 2 gene product (FIG. 55); a PIG37 gene product (FIG. 56); a GIG44 gene product (FIG. 57); and a GIG31 gene product

(FIG. 58) of the present invention, respectively.

FIG. 59, FIG. 60, FIG. 61, FIG. 67, FIG. 68, FIG. 69, FIG. 70, FIG. 71, FIG. 72, FIG. 73, FIG. 76, FIG. 82, FIG. 83, FIG. 86 and FIG. 87 are diagrams showing northern

blotting results that the GIG8 gene; the GIGlO gene; the GIGl 3 gene; the GIG30 gene;

the GIG32 gene; the GIG33 gene; the GIG34 gene; the GIG35 gene; the GIG38 gene;

the GIG39 gene; the GIG43 gene; the PIG49 gene; the PIG51 gene; the GIG44 gene; and the GIG31 gene are differentially expressed in a normal breast tissue, a primary

breast cancer tissue and a breast cancer cell line, respectively;

FIG. 62 is a diagram showing a northern blotting result that the GIG 15 gene of

the present invention is differentially expressed in a normal bone marrow tissue, a

leukemic bone marrow tissue, and a K562 leukemia cell;

FIG. 63, FIG. 64, FIG. 65, FIG. 66, FIG. 74, FIG. 75, FIG. 78, FIG. 79, FIG. 80

and FIG. 85 are diagrams showing northern blotting results that the GIG16 gene; the GIG24 gene; the GIG26 gene; the GIG29 gene; the GIG40 gene; the GIG42 gene; the

PIG33 gene; the PIG35 gene; the PIG36 gene; and the PIG37 gene are differentially

expressed in a normal liver tissue, a primary liver cancer tissue and a liver cancer cell

line, respectively;

FIG. 77 and FIG. 81 are diagrams showing northern blotting results that the GIG46 gene; and the MIG20 gene are differentially expressed in a normal exocervical

tissue, a primary uterine cancer tissue and a uterine cancer cell line, respectively;

FIG. 84 is a diagram showing a northern blotting result that the MIG 12 gene of

the present invention is differentially expressed in a normal lung tissue, a primary lung cancer tissue, a metastatic lung cancer tissue and a lung cancer cell line, and bottoms of FIGs. 59 to 87 are diagrams showing northern blotting results obtained by hybridizing

the same blots with β -actin probe, respectively. FIGs. 88 to 116 are diagrams showing northern blotting results that the GIG8

gene (FIG. 88); the GIGlO gene (FIG. 89); the GIGl 3 gene (FIG. 90); the GIGl 5 gene

(FIG. 91); the GIGl 6 gene (FIG. 92); the GIG24 gene (FIG. 93); the GIG26 gene (FIG.

94); the GIG29 gene (FIG. 95); the GIG30 gene (FIG. 96); the GIG32 gene (FIG. 97);

the GIG33 gene (FIG. 98); the GIG34 gene (FIG. 99); the GIG35 gene (FIG. 100); the

GIG38 gene (FIG. 101); the GIG39 gene (FIG. 102); the GIG40 gene (FIG. 103); the

GIG42 gene (FIG. 104); the GIG43 gene (FIG. 105); the GIG46 gene (FIG. 106); the

PIG33 gene (FIG. 107); the PIG35 gene (FIG. 108); the PIG36 gene (FIG. 109); the

MIG20 gene (FIG. 110); the PIG49 gene (FIG. I l l); the PIG51 gene (FIG. 112); the

MIG12 gene (FIG. 113); the PIG37 gene (FIG. 114); the GIG44 gene (FIG. 115); and

the GIG31 gene (FIG. 116) are differentially expressed in various normal tissues,

respectively, and bottoms of FIGs. 88 to 116 are diagrams showing northern blotting

results obtained by hybridizing the same blots with β -actin probe, respectively.

FIGs. 117 to 145 are diagrams showing northern blotting results that the GIG8

gene (FIG. 117); the GIGlO gene (FIG. 118); the GIGl 3 gene (FIG. 119); the GIGl 5 gene (FIG. 120); the GIG16 gene (FIG. 121); the GIG24 gene (FIG. 122); the GIG26

gene (FIG. 123); the GIG29 gene (FIG. 124); the GIG30 gene (FIG. 125); the GIG32

gene (FIG. 126); the GIG33 gene (FIG. 127); the GIG34 gene (FIG. 128); the GIG35 gene (FIG. 129); the GIG38 gene (FIG. 130); the GIG39 gene (FIG. 131); the GIG40

gene (FIG. 132); the GIG42 gene (FIG. 133); the GIG43 gene (FIG. 134); the GIG46 gene (FIG. 135); the PIG33 gene (FIG. 136); the PIG35 gene (FIG. 137); the PIG36 gene (FIG. 138); the MIG20 gene (FIG. 139); the PIG49 gene (FIG. 140); the PIG51

gene (FIG. 141); the MIGl 2 gene (FIG. 142); the PIG37 gene (FIG. 143); the GIG44 gene (FIG. 144); and the GIG31 gene (FIG. 145) are differentially expressed in various cancer cell lines, respectively, and bottoms of FIGs. 117 to 145 are diagrams showing

northern blotting results obtained by hybridizing the same blots with β -actin probe,

respectively.

FIG. 146, FIG. 147, FIG. 148, FIG. 154, FIG. 155, FIG. 156, FIG. 157, FIG. 158,

FIG. 159, FIG. 160, FIG. 169, FIG. 170, FIG. 173 and FIG. 174 are diagrams showing

growth curves of a wild-type MCF-7 cell; MCF-7 breast cancer cells transfected with

the GIG8 gene; the GIGlO gene; the GIGl 3 gene; the GIG30 gene; GIG32 gene; the

GIG33 gene; the GIG34 gene; the GIG35 gene; the GIG38 gene; the GIG39 gene; the PIG49 gene; the PIG33 gene; the GIG44 gene; and the GIG31 gene, respectively; and a

MCF-7 cell transfected with the expression vector pcDNA3.1, respectively;

FIG. 149 is a diagram showing growth curves of a wild-type K562 cell line; a

K562 leukemia cell transfected with the GIGl 5 gene; and a K562 cell transfected with

the expression vector pcDNA3.1 ;

FIG. 150, FIG. 151, FIG. 152, FIG. 153, FIG. 161, FIG. 162, FIG. 163, FIG. 165,

FIG. 166, FIG. 167 and FIG. 172 are diagrams showing growth curves of a wild-type HepG2 liver cancer cell line; HepG2 liver cancer cells transfected with the GIG 16 gene;

the GIG24 gene; the GIG26 gene; the GIG29 gene; the GIG40 gene; the GIG42 gene; the GIG43 gene; the PIG33 gene; the PIG35 gene; the PIG36 gene; and the PIG37 gene,

respectively; and a HepG2 cell transfected with the expression vector pcDN A3.1 ;

FIG. 164 and FIG. 168 are diagrams showing growth curves of a wild-type HeLa

cell; HeLa uterine cancer cells transfected with the GIG46 gene; and the MIG20 gene, respectively; and a HeLa cell transfected with the expression vector pcDNA3.1, respectively; and

FIG. 171 is a diagram showing growth curves of a wild-type A549 lung cancer

cell line; an A549 lung cancer cell transfected with the MIGl 2 gene; and an A549 cell

transfected with the expression vector pcDNA3.1.

BEST MODES FOR CARRYING OUT THE INVENTION

Hereinafter, preferred embodiments of the present invention will be described in

detail with reference to the accompanying drawings.

1. GIG8

The gene of the present invention is a human cancer suppressor gene 8 (GIG8)

having a DNA sequence of SEQ ID NO: 1, which was deposited with Accession No.

AY634687 into the GenBank database of U.S. National Institutes of Health (NIH)

(Scheduled Release Date: December 31, 2005), and a DNA sequence of the deposited

gene is similar to that of the Homo sapiens inhibitor of DNA binding 2, dominant

negative helix-loop-helix protein (ID2) gene deposited with Accession No. NM 002166

into the database. From this study result, it was however found that the GIG8 gene was closely related to various human carcinogenesis. From the study result, it was found that the GIG8 tumor suppressor gene was rarely expressed or not expressed in various human tumors including the breast cancer, while its expression was significantly

increased in various normal tissues.

The DNA sequence of SEQ ID NO: 1 has one open reading frame (ORF)

corresponding to base positions from 120 to 524 of the DNA sequence (base positions

from 522 to 524 represent a stop codon). A protein expressed from the gene of the present invention consists of 134

amino acid residues, and has an amino acid sequence of SEQ ID NO: 2 and a molecular

weight of approximately 15 kDa.

The gene and the protein of the present invention may be separated from human

tissues, or also be synthesized according to the known methods for synthesizing DNA or

peptide. For example, the gene of the present invention may be screened and cloned

according to the conventional methods on the basis of the information on the DNA sequence set forth in SEQ ID NO: 1. As another example, a 163-bp cDNA fragment,

which is not expressed or rarely expressed in the cancer tissue or the cancer cell line but

differentially expressed in the normal tissue, may be obtained by carrying out a reverse

transcription-polymerase chain reaction (RT-PCR) on the total RNA extracted from a

normal tissue, and a cancer tissue or a cancer cell line using a random primer H-AP33 of

SEQ ID NO: 3 (5'-AAGCTTGCTGCTC-S¹) and an anchored oligo-dT primer of SEQ

ID NO: 4 (5'-AAGCTTTTTTTTTTTA-3^l), and the resultant fragment, which is used as

the probe, may be plaque-hybridized with a cDNA library to obtain a full-length cDNA

clone.

It is regarded that the gene of the present invention is overexpressed in the

normal tissues, preferably the breast, the brain, the heart, the muscles, the large intestine, the thymus, the spleen, the kidney, the liver, the small intestine, the placenta and the

lungs to suppress the carcinogenesis. The gene of the present invention is mainly overexpressed in these tissues as an mRNA transcript having a size of approximately 1.3

kb.

2. GIGlO The gene of the present invention is a human cancer suppressor gene 10 (GIGlO)

having a DNA sequence of SEQ ID NO: 5, which was deposited with Accession No.

AY542305 into the GenBank database of U.S. National Institutes of Health (NIH) (Scheduled Release Date: December 31, 2005), and a DNA sequence of the deposited gene is similar to that of the Homo sapiens c-Cbl-interacting protein (CIN85) mRNA

gene deposited with Accession No. AF230904 into the database.

From this study result, it was however found that the GIGlO gene was closely

related to various human carcinogenesis. From the study result, it was found that the

GIGlO tumor suppressor gene was rarely expressed or not expressed in various human

tumors including the breast cancer, while its expression was significantly increased in

various normal tissues.

The DNA sequence of SEQ ID NO: 5 has one open reading frame (ORF)

corresponding to base positions from 52 to 2,049 of the DNA sequence (base positions

from 2,047 to 2,049 represent a stop codon). A protein expressed from the gene of the present invention consists of 665

amino acid residues, and has an amino acid sequence of SEQ ID NO: 6 and a molecular

weight of approximately 73 kDa.

The gene and the protein of the present invention may be separated from human

peptide. For example, the gene of the present invention may be screened and cloned according to the conventional methods on the basis of the information on the DNA sequence set forth in SEQ ID NO: 5. As another example, a 321-bp cDNA fragment, which is not expressed or rarely expressed in the cancer tissue or the cancer cell line but differentially expressed in the normal tissue, may be obtained by carrying out a reverse

normal tissue, and a cancer tissue or a cancer cell line using a random primer H-APlO of

SEQ ID NO: 7 (5'-AAGCTTCCACGTA-S') and an anchored oligo-dT primer of SEQ

ID NO: 8 (5'-AAGCTTTTTTTTTTTC-S'), and the resultant fragment, which is used as

the probe, may be plaque-hybridized with a cDNA library to obtain a full-length cDNA clone.

It is regarded that the gene of the present invention is overexpressed in the

normal tissues, preferably the breast, the brain, the heart, the muscles, the thymus, the

spleen, the kidney, the liver, the small intestine, the placenta and the lungs to suppress

the carcinogenesis. The gene of the present invention is mainly overexpressed in these

tissues as an mRNA transcript having a size of approximately 3.5 kb. 3. GIGl 3

The gene of the present invention is a human cancer suppressor gene 13 (GIG13)

having a DNA sequence of SEQ ID NO: 9, which was deposited with Accession No.

AY493418 into the GenBank database of U.S. National Institutes of Health (NIH)

(Scheduled Release Date: December 31, 2005), and a DNA sequence of the deposited gene is similar to that of the Homo sapiens period homolog 3 (Drosophila) (PER3) gene

deposited with Accession No. NM 016831 into the database.

From this study result, it was however found that the GIG 13 gene was closely related to various human carcinogenesis. From the study result, it was found that the

GIG 13 tumor suppressor gene was rarely expressed or not expressed in various human

tumors including the breast cancer, while its expression was significantly increased in various normal tissues. The DNA sequence of SEQ ID NO: 9 has one open reading

frame (ORF) corresponding to base positions from 72 to 3,677 of the DNA sequence (base positions from 3,675 to 3,677 represent a stop codon).

A protein expressed from the gene of the present invention consists of 1,201

amino acid residues, and has an amino acid sequence of SEQ ID NO: 10 and a

molecular weight of approximately 132 kDa.

The gene and the protein of the present invention may be separated from human

peptide. For example, the gene of the present invention may be screened and cloned according to the conventional methods on the basis of the information on the DNA sequence set forth in SEQ ID NO: 1. As another example, a 347-bp cDNA fragment,

differentially expressed in the normal tissue, may be obtained by carrying out a reverse transcription-polymerase chain reaction (RT-PCR) on the total RNA extracted from a

normal tissue, and a cancer tissue or a cancer cell line using a random primer H-AP5 of

SEQ ID NO: 11 (5'-AAGCTTAGTAGGC-S¹) and an anchored oligo-dT primer of SEQ

ID NO: 12 (5^I-AAGCTTTTTTTTTTTC-3^I), and the resultant fragment, which is used as the probe, may be plaque-hybridized with a cDNA library to obtain a full-length cDNA

clone. It is regarded that the gene of the present invention is overexpressed in the normal tissues, preferably the breast and the liver to suppress the carcinogenesis. The

gene of the present invention is mainly overexpressed in these tissues as an mRNA

transcript having a size of approximately 1.3 kb. Especially, the gene of the present invention is differentially expressed only in the normal tissues. For example, the gene

of the present invention is rarely expressed or not expressed in the cancer tissues and the

cancer cells such as the breast cancer tissue, and the breast cancer cell line MCF-7, but

differentially increasingly expressed only in the normal breast tissues.

4. GIGl 5

The gene of the present invention is a human cancer suppressor gene 15 (GIGl 5)

having a DNA sequence of SEQ ID NO: 13, which was deposited with Accession No.

AY927233 into the GenBank database of U.S. National Institutes of Health (NIH)

(Scheduled Release Date: October 1, 2006), and a DNA sequence of the deposited gene

is similar to that of the Homo sapiens chromosome 11, clone RP11-466H18 gene deposited with Accession No. ACl 16533 into the database. From this study result, it was however found that the GIGl 5 gene was closely related to various human

carcinogenesis. From the study result, it was found that the GIG 15 tumor suppressor

gene was rarely expressed or not expressed in various human tumors including the leukemia, while its expression was significantly increased in various normal tissues.

The DNA sequence of SEQ ID NO: 13 has one open reading frame (ORF) corresponding to base positions from 18 to 338 of the DNA sequence (base positions

from 336 to 338 represent a stop codon).

A protein expressed from the gene of the present invention consists of 106

amino acid residues, and has an amino acid sequence of SEQ ID NO: 14 and a molecular weight of approximately 12 kDa.

The gene and the protein of the present invention may be separated from human tissues, or also be synthesized according to the known methods for synthesizing DNA or peptide. For example, the gene of the present invention may be screened and cloned

according to the conventional methods on the basis of the information on the DNA

sequence set forth in SEQ ID NO: 13. As another example, a 133-bp cDNA fragment,

which is rarely expressed in the cancer tissue or the cancer cell line but differentially

expressed in the normal tissue, may be obtained by carrying out a reverse

transcription-polymerase chain reaction (RT-PCR) on the total RNA extracted from a normal tissue, and a cancer tissue or a cancer cell line using a random primer H-AP2 of

SEQ ID NO: 15 (5'-AAGCTTCGACTGT-S¹) and an anchored oligo-dT primer of SEQ

ID NO: 16 (5'-AAGCTTTTTTTTTTTC-3^l), and the resultant fragment, which is used as

clone.

It is regarded that the gene of the present invention is overexpressed in the

lungs to suppress the carcinogenesis. The gene of the present invention is mainly overexpressed in these tissues as an mRNA transcript having a size of approximately 0.5

kb. Especially, the gene of the present invention is differentially expressed only in the normal tissues. For example, the gene of the present invention is rarely expressed in the cancer tissues and the cancer cells such as the leukemia cell and the leukemia cell

line K562, but differentially increasingly expressed only in the normal breast tissues.

5. GIGl 6

The gene of the present invention is a human cancer suppressor gene 16 (GIG 16)

having a DNA sequence of SEQ ID NO: 17, which was deposited with Accession No. AY513277 into the GenBank database of U.S. National Institutes of Health (NIH)

(Scheduled Release Date: December 31, 2005), and some DNA sequence of the

deposited gene is different from that of the Homo sapiens hydroxyacid oxidase 2 (long

chain) (HAO2) gene deposited with Accession No. NM_016527 into the database, the

HA02 gene being known to be one of three genes having 2-hydroxyacid oxidase

activity (Jones, J.M., et al., J. Biol. Chem. 275(17), 12590-12597 (2000)). From this

study result, it was however found that the GIG 16 tumor suppressor gene was not

expressed at all in various human tumors including the liver cancer, while its expression

was significantly increased in various normal tissues.

The DNA sequence of SEQ ID NO: 17 has one open reading frame (ORF)

corresponding to base positions from 41 to 1,096 of the DNA sequence (base positions

from 1,094 to 1,096 represent a stop codon).

A protein expressed from the gene of the present invention consists of 351

amino acid residues, and has an amino acid sequence of SEQ ID NO: 18 and a molecular weight of approximately 39 kDa.

The gene and the protein of the present invention may be separated from human

according to the conventional methods on the basis of the information on the DNA sequence set forth in SEQ ID NO: 17. As another example, a 213-bp cDNA fragment, which is not expressed in the cancer tissue or the cancer cell line but differentially expressed in the normal tissue, may be obtained by carrying out a reverse transcription-polymerase chain reaction (RT-PCR) on the total RNA extracted from a normal tissue, and a cancer tissue or a cancer cell line using a random primer H-AP8 of

SEQ ID NO: 19 (5'-AAGCTTTTACCGC-S¹) and an anchored oligo-dT primer of SEQ

ID NO: 20 (5'-AAGCTTTTTTTTTTTC-S'), and the resultant fragment, which is used as

the probe, may be plaque-hybridized with a cDNA library to obtain a full-length cDNA clone. The present inventors inserted the full-length GIG 16 cDNA into the expression

vector pBAD/Thio-Topo (Invitrogen, U.A.), and then transformed E. coli DH5 α with

the resultant expression vector to obtain a transformant, which was designated E. coli

DH5 α /GIG16/pBAD/Thio-Topo.

It is regarded that the gene of the present invention is overexpressed in the

normal tissues, preferably the liver and the kidney to suppress the carcinogenesis. Also,

it is regarded that its gene expression is suppressed in the leukemia, the uterine cancer, the malignant lymphoma, the colon cancer, the lung cancer and the skin cancer to induce

tissues as an mRNA transcript having a size of approximately 2.0 kb. Especially, the

gene of the present invention is differentially expressed only in the normal tissues. For

example, the gene of the present invention is not expressed in the cancer tissues and the cancer cells such as the liver cancer tissue, and the liver cancer cell line HepG2, but differentially expressed only in the normal breast tissues. 6. GIG24

The gene of the present invention is a human cancer suppressor gene 24 (GIG24) having a DNA sequence of SEQ ID NO: 21, which was deposited with Accession No. AY513275 into the GenBank database of U.S. National Institutes of Health (NIH) (Scheduled Release Date: December 31, 2005), and a DNA sequence of the deposited gene is similar to that of the Homo sapiens cDNA FLJ35730 fis, clone TESTI2003131,

highly similar to ALPHA- 1-ANTICHYMOTRYPSIN PRECURSOR gene deposited

with Accession No. AK093049 into the database. From this study result, it was

however found that the GIG24 tumor suppressor gene was not expressed at all in

various human tumors including the liver cancer, while its expression was significantly

increased in the normal liver tissue.

The DNA sequence of SEQ ID NO: 21 has one open reading frame (ORF)

corresponding to base positions from 34 to 1,305 of the DNA sequence (base positions

from 1,303 to 1,305 represent a stop codon).

A protein expressed from the gene of the present invention consists of 423

amino acid residues, and has an amino acid sequence of SEQ ID NO: 22 and a

molecular weight of approximately 47 kDa.

The gene and the protein of the present invention may be separated from human

according to the conventional methods on the basis of the information on the DNA sequence set forth in SEQ ID NO: 1. As another example, a 221-bp cDNA fragment, which is not expressed in the cancer tissue or the cancer cell line but differentially

expressed in the normal tissue, may be obtained by carrying out a reverse transcription-polymerase chain reaction (RT-PCR) on the total RNA extracted from a normal tissue, and a cancer tissue or a cancer cell line using a random primer H-AP7 of

SEQ ID NO: 23 (5'-AAGCTTAACGAGG-S') and an anchored oligo-dT primer of SEQ

ID NO: 24 (5^t-AAGCTTTTTTTTTTTC-3^l), and the resultant fragment, which is used as the probe, may be plaque-hybridized with a cDNA library to obtain a full-length cDNA

clone. The present inventors inserted the full-length GIG24 cDNA into the expression

DH5 α /GIG24/pBAD/Thio-Topo.

It is regarded that the gene of the present invention is overexpressed in the

normal tissues, preferably the liver, the heart and the muscles to suppress the

carcinogenesis. Also, it is regarded that its gene expression is suppressed in the leukemia, the uterine cancer, the malignant lymphoma, the colon cancer, the lung cancer

and the skin cancer to induce the carcinogenesis. The gene of the present invention is

mainly overexpressed in these tissues as an mRNA transcript having a size of

approximately 2.4 kb.

7. GIG26

The gene of the present invention is a human cancer suppressor gene 26 (GIG26) having a DNA sequence of SEQ ID NO: 25, which was deposited with Accession No.

AY544126 into the GenBank database of U.S. National Institutes of Health (NIH)

gene is similar to those of the Homo sapiens prostate-specific membrane antigen-like

(PSMAL) gene and the Homo sapiens prostate-specific membrane antigen-like protein

(PSMAL/GCP III) mRNA gene, deposited with Accession No. NM_153696 and

AF261715 into the database, respectively, the prostate-specific membrane antigen being

known to be a marker protein that is mainly expressed in a prostate epithelium ((Lee,

S.J., et al., J. MoI. Biol. 330(4), 749-760 (2003)). From this study result, it was however found that the GIG26 tumor suppressor gene was not expressed at all in various human tumors including the liver cancer, while its expression was significantly

increased in various normal tissues.

The DNA sequence of SEQ ID NO: 25 has one open reading frame (ORF)

corresponding to base positions from 26 to 1,354 of the DNA sequence (base positions

from 1,352 to 1,354 represent a stop codon).

A protein expressed from the gene of the present invention consists of 442

amino acid residues, and has an amino acid sequence of SEQ ID NO: 26 and a molecular weight of approximately 50 kDa. The gene and the protein of the present invention may be separated from human

sequence set forth in SEQ ID NO: 25. As another example, a 204-bp cDNA fragment,

which is not expressed in the cancer tissue or the cancer cell line but differentially expressed in the normal tissue, may be obtained by carrying out a reverse transcription-polymerase chain reaction (RT-PCR) on the total RNA extracted from a

normal tissue, and a cancer tissue or a cancer cell line using a random primer H-APl 1 of SEQ ID NO: 27 (5'-AAGCTTCGGGTAA-S') and an anchored oligo-dT primer of SEQ

ID NO: 28 (5'-AAGCTTTTTTTTTTTG-S'), and the resultant fragment, which is used as the probe, may be plaque-hybridized with a cDNA library to obtain a full-length cDNA

clone.

The present inventors inserted the full-length GIG26 cDNA into the expression vector pBAD/Thio-Topo (Invitrogen, U.A.), and then transformed E. coli DH5 α with

DH5 α /GIG26/pBAD/Thio-Topo.

It is regarded that the gene of the present invention is overexpressed in the

normal tissues, preferably the liver, the kidney, the brain and the heart to suppress the carcinogenesis. Also, it is regarded that its gene expression is suppressed in leukemia,

uterine cancer, malignant lymphoma, colon cancer, lung cancer and skin cancer to

induce the carcinogenesis. The gene of the present invention is mainly overexpressed

in these tissues as an mRNA transcript having a size of approximately 2.0 kb.

8. GIG29

The gene of the present invention is a human cancer suppressor gene 29 (GIG29)

having a DNA sequence of SEQ ID NO: 29, which was deposited with Accession No.

AY544127 into the GenBank database of U.S. National Institutes of Health (NIH)

(Scheduled Release Date: December 31, 2005), and a DNA sequence of the deposited gene is similar to that of the Homo sapiens solute carrier family 10 (sodium/bile acid

cotransporter family) gene deposited with Accession No. NM 003049 into the database.

From this study result, it was however found that the GIG29 tumor suppressor gene was not expressed at all in various human tumors including the liver cancer, while its

expression was significantly increased in the normal liver tissue.

The DNA sequence of SEQ ID NO: 29 has one open reading frame (ORF) corresponding to base positions from 62 to 1,111 of the DNA sequence (base positions

from 1,109 to 1,111 represent a stop codon).

A protein expressed from the gene of the present invention consists of 349 amino acid residues, and has an amino acid sequence of SEQ ID NO: 30 and a

molecular weight of approximately 38 kDa.

The gene and the protein of the present invention may be separated from human

according to the conventional methods on the basis of the information on the DNA sequence set forth in SEQ ID NO: 29. As another example, a 277-bp cDNA fragment,

which is not expressed in the cancer tissue or the cancer cell line but differentially

expressed in the normal tissue, may be obtained by carrying out a reverse

transcription-polymerase chain reaction (RT-PCR) on the total RNA extracted from a normal tissue, and a cancer tissue or a cancer cell line using a random primer H-AP3 of SEQ ID NO: 31 (5'-AAGCTTTGGTCAG-S¹) and an anchored oligo-dT primer of SEQ

ID NO: 32 (5'-AAGCTTTTTTTTTTTA-3¹), and the resultant fragment, which is used as the probe, may be plaque-hybridized with a cDNA library to obtain a full-length cDNA

clone. The present inventors inserted the full-length GIG29 cDNA into the expression

DH5 α /GIG29/pBAD/Thio-Topo.

It is regarded that the gene of the present invention is overexpressed in the normal tissues, preferably the liver to suppress the carcinogenesis. Also, it is regarded that its gene expression is suppressed in the leukemia, the uterine cancer, the malignant lymphoma, the colon cancer, the lung cancer and the skin cancer to induce the

carcinogenesis. The gene of the present invention is mainly overexpressed in these tissues as an mRNA transcript having a size of approximately 1.4 kb.

9. GIG30

The gene of the present invention is a human cancer suppressor gene 30 (GIG30)

having a DNA sequence of SEQ ID NO: 33, which was deposited with Accession No.

AY524045 into the GenBank database of U.S. National Institutes of Health (NIH)

gene is similar to that of the Homo sapiens clone DNA43305 RIPK2 (UNQ277) gene deposited with Accession No. AY358814 into the database. From this study result, it

was however found that the GIG30 gene was closely related to various human

carcinogenesis. From the study result, it was found that the GIG30 tumor suppressor

gene was rarely expressed or not expressed in various human tumors including the

breast cancer, while its expression was significantly increased in various normal tissues.

The DNA sequence of SEQ ID NO: 33 has one open reading frame (ORF)

corresponding to base positions from 88 to 1,710 of the DNA sequence (base positions

from 1 ,708 to 1,710 represent a stop codon).

A protein expressed from the gene of the present invention consists of 540

amino acid residues, and has an amino acid sequence of SEQ ID NO: 34 and a

molecular weight of approximately 61 kDa.

The gene and the protein of the present invention may be separated from human

tissues, or also be synthesized according to the known methods for synthesizing DNA or peptide. For example, the gene of the present invention may be screened and cloned

sequence set forth in SEQ ID NO: 35. As another example, a 278-bp cDNA fragment, which is not expressed or rarely expressed in the cancer tissue or the cancer cell line but

normal tissue, and a cancer tissue or a cancer cell line using a random primer H-AP4 of SEQ ID NO: 35 (5'-AAGCTTCTCAACG-S¹) and an anchored oligo-dT primer of SEQ

ID NO: 36 (5^t-AAGCTTTTTTTTTTTG-3¹), and the resultant fragment, which is used as

It is regarded that the gene of the present invention is overexpressed in the

normal tissues, preferably the breast, the heart, the muscles and the liver to suppress the

carcinogenesis. The gene of the present invention is mainly overexpressed in these

tissues as an mRNA transcript having a size of approximately 1.9 kb.

10. GIG32

The gene of the present invention is a human cancer suppressor gene 32 (GIG32)

having a DNA sequence of SEQ ID NO: 37, which was deposited with Accession No. AY762103 into the GenBank database of U.S. National Institutes of Health (NIH)

(Scheduled Release Date: December 31, 2005), and a DNA sequence of the deposited gene is similar to that of the Homo sapiens caveolin 1, caveolae protein, 22 kDa

(CAVl) gene deposited with Accession No. NM OO 1753 into the database. From this

study result, it was however found that the GIG32 gene was closely related to various human carcinogenesis. From the study result, it was found that the GIG32 tumor

suppressor gene was rarely expressed or not expressed in various human tumors including the breast cancer, while its expression was significantly increased in various normal tissues.

The DNA sequence of SEQ ID NO: 37 has one open reading frame (ORF)

corresponding to base positions from 43 to 579 of the DNA sequence (base positions

from 577 to 579 represent a stop codon).

A protein expressed from the gene of the present invention consists of 178

amino acid residues, and has an amino acid sequence of SEQ ID NO: 38 and a

molecular weight of approximately 20 kDa.

The gene and the protein of the present invention may be separated from human

according to the conventional methods on the basis of the information on the DNA sequence set forth in SEQ ID NO: 37. As another example, a 172-bp cDNA fragment,

transcription-polymerase chain reaction (RT-PCR) on the total RNA extracted from a normal tissue, and a cancer tissue or a cancer cell line using a random primer H-AP8 of SEQ ID NO: 39 (5'-AAGCTTTTACCGC-S') and an anchored oligo-dT primer of SEQ ID NO: 40 (5¹-AAGCTTTTTTTTTTTG-3^l), and the resultant fragment, which is used as

clone.

It is regarded that the gene of the present invention is overexpressed in the

normal tissues, preferably the breast, the brain, the heart, the muscles, the large intestine,

the thymus, the spleen, the kidney, the liver, the small intestine, the placenta and the lungs to suppress the carcinogenesis. The gene of the present invention is mainly

overexpressed in these tissues as an mRNA transcript having a size of approximately 4.0 kb.

11. GIG33

The gene of the present invention is a human cancer suppressor gene 33 (GIG33)

having a DNA sequence of SEQ ID NO: 41, which was deposited with Accession No. AY871273 into the GenBank database of U.S. National Institutes of Health (NIH) (Scheduled Release Date: October 1, 2006), and a DNA sequence of the deposited gene

is similar to that of the Homo sapiens ribosomal protein L35a (RPL35A) gene deposited

with Accession No. NM_000996 into the database. A ribosomal gene is an

intracellular organelle that catalyzes the protein synthesis and consists of a small 4OS

subunit and a large 60S subunit. However, a function of the gene remains to be specified (Herzog, H., et al., Nucleic Acids Res., 18(15), 4600 (1990); Kenmochi, N., et

al., Genome Res., 8(5), 509-523 (1998); Lopez, CD., et al., Cancer Lett. 180(2),

195-202 (2002)). From this study result, it was however found that the GIG33 gene

was closely related to various human carcinogenesis. From the study result, it was found that the GIG33 tumor suppressor gene was rarely expressed or not expressed in

various human tumors including the breast cancer, while its expression was significantly

increased in various normal tissues. The DNA sequence of SEQ ID NO: 41 has one open reading frame (ORF) corresponding to base positions from 74 to 406 of the DNA sequence (base positions

from 404 to 406 represent a stop codon).

A protein expressed from the gene of the present invention consists of 110 amino acid residues, and has an amino acid sequence of SEQ ID NO: 42 and a

molecular weight of approximately 12 kDa.

The gene and the protein of the present invention may be separated from human

sequence set forth in SEQ ID NO: 41. As another example, a 182-bp cDNA fragment,

which is not expressed or rarely expressed in the cancer tissue or the cancer cell line but differentially expressed in the normal tissue, may be obtained by carrying out a reverse

SEQ ID NO: 43 (5'-AAGCTTGCTGCTC-S¹) and an anchored oligo-dT primer of SEQ

ID NO: 44 (5'-AAGCTTTTTTTTTTTG-S'), and the resultant fragment, which is used as the probe, may be plaque-hybridized with a cDNA library to obtain a full-length cDNA

clone.

It is regarded that the gene of the present invention is overexpressed in the

the thymus, the spleen, the kidney, the liver, the small intestine, the placenta, the lungs

and the peripheral blood leukocyte to suppress the carcinogenesis. The gene of the present invention is mainly overexpressed in these tissues as an mRNA transcript having a size of approximately 0.6 kb.

12. GIG34 The gene of the present invention is a human cancer suppressor gene 34 (GIG34) having a DNA sequence of SEQ ID NO: 45, which was deposited with Accession No.

AY871274 into the GenBank database of U.S. National Institutes of Health (NIH) (Scheduled Release Date: October 1, 2006), and a DNA sequence of the deposited gene

is similar to that of the Homo sapiens ribosomal protein Ll 1 gene deposited with

Accession No. BCO 18970 into the database. From this study result, it was however

found that the GIG34 gene was closely related to various human carcinogenesis. From

the study result, it was found that the GIG34 tumor suppressor gene was rarely expressed or not expressed in various human tumors including the breast cancer, while

its expression was significantly increased in various normal tissues. The DNA sequence of SEQ ID NO: 45 has one open reading frame (ORF) corresponding to base positions from 5 to 538 of the DNA sequence (base positions

from 536 to 538 represent a stop codon).

A protein expressed from the gene of the present invention consists of 177

amino acid residues, and has an amino acid sequence of SEQ ID NO: 46 and a

molecular weight of approximately 20 kDa.

The gene and the protein of the present invention may be separated from human tissues, or also be synthesized according to the known methods for synthesizing DNA or

according to the conventional methods on the basis of the information on the DNA sequence set forth in SEQ ID NO: 45. As another example, a 205-bp cDNA fragment, which is not expressed or rarely expressed in the cancer tissue or the cancer cell line but

differentially expressed in the normal tissue, may be obtained by carrying out a reverse transcription-polymerase chain reaction (RT-PCR) on the total RNA extracted from a normal tissue, and a cancer tissue or a cancer cell line using a random primer H-AP35 of

SEQ ID NO: 47 (S'-AAGCTTCAGGGCA-S¹) and an anchored oligo-dT primer of SEQ

ID NO: 48 (5^t-AAGCTTTTTTTTTTTC-3^l), and the resultant fragment, which is used as

It is regarded that the gene of the present invention is overexpressed in the

and the peripheral blood leukocyte to suppress the carcinogenesis. The gene of the

present invention is mainly overexpressed in these tissues as an mRNA transcript having

a size of approximately 0.6 kb. 13. GIG35

The gene of the present invention is a human cancer suppressor gene 35 (GIG35)

having a DNA sequence of SEQ ID NO: 49, which was deposited with Accession No.

AY542307 into the GenBank database of U.S. National Institutes of Health (NIH)

gene is similar to that of the Homo sapiens eukaryotic translation elongation factor 1 gamma (EEFlG) gene deposited with Accession No. NM OO 1404 into the database. The gene is a subunit of the elongation factor 1 that takes an important role in

transferring aminoacyl tRNAs to ribosome ((Kumabe, T., et al., Nucleic Acids Res., 20(10), 2598 (1992)). From this study result, it was however found that the GIG35

gene was closely related to various human carcinogenesis. From the study result, it

was found that the GIG35 tumor suppressor gene was rarely expressed in various human tumors including the breast cancer, while its expression was significantly increased in various normal tissues.

The DNA sequence of SEQ ID NO: 49 has one open reading frame (ORP)

corresponding to base positions from 19 to 1,332 of the DNA sequence (base positions from 1,330 to 1,332 represent a stop codon).

A protein expressed from the gene of the present invention consists of 437

amino acid residues, and has an amino acid sequence of SEQ ID NO: 50 and a

molecular weight of approximately 50 kDa.

sequence set forth in SEQ ID NO: 49. As another example, a 212-bp cDNA fragment,

transcription-polymerase chain reaction (RT-PCR) on the total RNA extracted from a normal tissue, and a cancer tissue or a cancer cell line using a random primer H-AP3 of

SEQ ID NO: 51 (5'-AAGCTTTGGTCAG-S¹) and an anchored oligo-dT primer of SEQ

ID NO: 52 (5^•-AAGCTTTTTTTTTTTC-3^l), and the resultant fragment, which is used as the probe, may be plaque-hybridized with a cDNA library to obtain a full-length cDNA clone.

It is regarded that the gene of the present invention is overexpressed in the

lungs to suppress the carcinogenesis. The gene of the present invention is mainly

overexpressed in these tissues as an mRNA transcript having a size of approximately 1.3 kb.

14. GIG38

The gene of the present invention is a human cancer suppressor gene 38 (GIG38)

having a DNA sequence of SEQ ID NO: 53, which was deposited with Accession No.

AY550970 into the GenBank database of U.S. National Institutes of Health (NIH)

gene is similar to that of the Homo sapiens sin3 -associated polypeptide, 18 kDa

(SAPl 8) gene deposited with Accession No. NM 005870 into the database. From this

study result, it was however found that the GIG38 gene was closely related to various

human carcinogenesis. From the study result, it was found that the GIG38 tumor

suppressor gene was rarely expressed or not expressed in various human tumors including the breast cancer, while its expression was significantly increased in various

normal tissues.

The DNA sequence of SEQ ID NO: 53 has one open reading frame (ORF) corresponding to base positions from 17 to 478 of the DNA sequence (base positions

from 476 to 478 represent a stop codon). A protein expressed from the gene of the present invention consists of 153 amino acid residues, and has an amino acid sequence of SEQ ID NO: 54 and a

molecular weight of approximately 17 kDa.

according to the conventional methods on the basis of the information on the DNA sequence set forth in SEQ ID NO: 53. As another example, a 172-bp cDNA fragment,

normal tissue, and a cancer tissue or a cancer cell line using a random primer H-AP 12 of

SEQ ID NO: 55 (5'-AAGCTTGAGTGCT-S') and an anchored oligo-dT primer of SEQ

ID NO: 56 (5'-AAGCTTTTTTTTTTTC-S'), and the resultant fragment, which is used as the probe, may be plaque-hybridized with a cDNA library to obtain a full-length cDNA

clone.

It is regarded that the gene of the present invention is overexpressed in the

normal tissues, preferably the breast, the heart, the muscles, the kidney, the liver and the placenta to suppress the carcinogenesis. The gene of the present invention is mainly overexpressed in these tissues as an mRNA transcript having a size of approximately 0.7

kb.

15. GIG39

The gene of the present invention is a human cancer suppressor gene 39 (GIG39) having a DNA sequence of SEQ ID NO: 57, which was deposited with Accession No.

AY550972 into the GenBank database of U.S. National Institutes of Health (NIH)

(Scheduled Release Date: December 31, 2005), and a DNA sequence of the deposited gene is similar to that of the Homo sapiens chromosome 1 open reading frame 24 gene deposited with Accession No. BC030531 into the database. From this study result, it

was however found that the GIG39 gene was closely related to various human

carcinogenesis. From the study result, it was found that the GIG39 tumor suppressor

breast cancer, while its expression was significantly increased in various normal tissues. The DNA sequence of SEQ ID NO: 57 has one open reading frame (ORP)

corresponding to base positions from 70 to 2,856 of the DNA sequence (base positions

from 2,854 to 2,856 represent a stop codon).

A protein expressed from the gene of the present invention consists of 928

amino acid residues, and has an amino acid sequence of SEQ ID NO: 58 and a

molecular weight of approximately 103 kDa.

according to the conventional methods on the basis of the information on the DNA sequence set forth in SEQ ID NO: 57. As another example, a 327-bp cDNA fragment, which is not expressed or rarely expressed in the cancer tissue or the cancer cell line but

transcription-polymerase chain reaction (RT-PCR) on the total RNA extracted from a normal tissue, and a cancer tissue or a cancer cell line using a random primer H-AP 12 of SEQ ID NO: 59 (5'-AAGCTTGAGTGCT-S¹) and an anchored oligo-dT primer of SEQ

ID NO: 60 (5^I-AAGCTTTTTTTTTTTA-3^I), and the resultant fragment, which is used as

It is regarded that the gene of the present invention is overexpressed in the

normal tissues, preferably the breast and the liver to suppress the carcinogenesis. The

gene of the present invention is mainly overexpressed in these tissues as an mRNA transcript having a size of approximately 2.4 kb.

16. GIG40

The gene of the present invention is a human cancer suppressor gene 40 (GIG40) having a DNA sequence of SEQ ID NO: 61, which was deposited with Accession No.

AY550966 into the GenBank database of U.S. National Institutes of Health (NIH)

gene is similar to that of the Homo sapiens epidermal growth factor receptor

(erythroblastic leukemia viral (v-erb-b) oncogene homolog, avian) (EGFR) deposited with Accession No. NM 005228 into the database. From this study result, it was

however found that the GIG40 tumor suppressor gene was rarely expressed in various

human tumors including the liver cancer, while its expression was significantly

increased in the normal liver tissue.

The DNA sequence of SEQ ID NO: 61 has one open reading frame (ORF)

corresponding to base positions from 47 to 3,679 of the DNA sequence (base positions

from 3,677 to 3,679 represent a stop codon). A protein expressed from the gene of the present invention consists of 1,210 amino acid residues, and has an amino acid sequence of SEQ ID NO: 62 and a

molecular weight of approximately 134 kDa.

peptide. For example, the gene of the present invention may be screened and cloned according to the conventional methods on the basis of the information on the DNA

sequence set forth in SEQ ID NO: 61. As another example, a 275-bp cDNA fragment,

expressed in the normal tissue, may be obtained by carrying out a reverse

transcription-polymerase chain reaction (RT-PCR) on the total RNA extracted from a normal tissue, and a cancer tissue or a cancer cell line using a random primer H-AP7 of

SEQ ID NO: 63 (5'-AAGCTTAACGAGG-S¹) and an anchored oligo-dT primer of SEQ

ID NO: 64 (5^I-AAGCTTTTTTTTTTTG-3^I), and the resultant fragment, which is used as

clone. The present inventors inserted the full-length GIG40 cDNA into the expression

DH5 α /GIG40/pB AD/Thio-Topo.

It is regarded that the gene of the present invention is overexpressed in the

normal tissues, preferably the liver, the heart and the muscles to suppress the

carcinogenesis. Also, it is regarded that its gene expression is suppressed in the leukemia, the uterine cancer, the malignant lymphoma, the colon cancer, the lung cancer and the skin cancer to induce the carcinogenesis. The gene of the present invention is mainly overexpressed in these tissues as an niRNA transcript having a size of

approximately 1.5 kb. 17. GIG42 The gene of the present invention is a human cancer suppressor gene 42 (GIG42) having a DNA sequence of SEQ ID NO: 65, which was deposited with Accession No.

AY550967 into the GenBank database of U.S. National Institutes of Health (NIH)

gene is similar to that of the Homo sapiens albumin gene deposited with Accession No.

BC034023 into the database. From this study result, it was however found that the

GIG42 tumor suppressor gene was not expressed at all in various human tumors

including the liver cancer, while its expression was significantly increased in various normal tissues. The DNA sequence of SEQ ID NO: 65 has one open reading frame (ORF)

corresponding to base positions from 8 to 1,837 of the DNA sequence (base positions

from 1,835 to 1,837 represent a stop codon).

A protein expressed from the gene of the present invention consists of 609

amino acid residues, and has an amino acid sequence of SEQ ID NO: 66 and a

molecular weight of approximately 69 kDa.

The gene and the protein of the present invention may be separated from human

sequence set forth in SEQ ID NO: 65. As another example, a 327-bp cDNA fragment, which is not expressed in the cancer tissue or the cancer cell line but differentially expressed in the normal tissue, may be obtained by carrying out a reverse transcription-polymerase chain reaction (RT-PCR) on the total RNA extracted from a normal tissue, and a cancer tissue or a cancer cell line using a random primer H-AP8 of

SEQ ID NO: 67 (5'-AAGCTTTTACCGC-S¹) and an anchored oligo-dT primer of SEQ

ID NO: 68 (5'-AAGCTTTTTTTTTTTG-S'), and the resultant fragment, which is used as the probe, may be plaque-hybridized with a cDNA library to obtain a full-length cDNA

clone. The present inventors inserted the full-length GIG42 cDNA into the expression

DH5 α /GIG42/pBAD/Thio-Topo.

It is regarded that the gene of the present invention is overexpressed in the normal tissues, preferably the liver to suppress the carcinogenesis. Also, it is regarded

that its gene expression is suppressed in the leukemia, the uterine cancer, the malignant lymphoma, the colon cancer, the lung cancer and the skin cancer to induce the

tissues as an mRNA transcript having a size of approximately 2.5 kb.

18. GIG43

The gene of the present invention is a human cancer suppressor gene 43 (GIG43) having a DNA sequence of SEQ ID NO: 69, which was deposited with Accession No. AY550971 into the GenBank database of U.S. National Institutes of Health (NIH)

gene is similar to that of the Homo sapiens phospholipid scramblase 4 gene deposited with Accession No. BC028354 into the database. From this study result, it was

however found that the GIG43 gene was closely related to various human

carcinogenesis. From the study result, it was found that the GIG43 tumor suppressor gene was rarely expressed or not expressed in various human tumors including the

The DNA sequence of SEQ ID NO: 69 has one open reading frame (ORF)

corresponding to base positions from 96 to 1 ,085 of the DNA sequence (base positions

from 1,083 to 1,085 represent a stop codon).

A protein expressed from the gene of the present invention consists of 329 amino acid residues, and has an amino acid sequence of SEQ ID NO: 70 and a

molecular weight of approximately 37 kDa.

The gene and the protein of the present invention may be separated from human

sequence set forth in SEQ ID NO: 69. As another example, a 273-bp cDNA fragment,

SEQ ID NO: 71 (5'-AAGCTTCCACGTA-S¹) and an anchored oligo-dT primer of SEQ

ID NO: 72 (5'-AAGCTTTTTTTTTTTG-S'), and the resultant fragment, which is used as the probe, may be plaque-hybridized with a cDNA library to obtain a full-length cDNA clone.

It is regarded that the gene of the present invention is overexpressed in the

normal tissues, preferably the breast, the heart, the kidney, the liver, the placenta and the lungs to suppress the carcinogenesis. The gene of the present invention is mainly

overexpressed in these tissues as an mRNA transcript having a size of approximately 3.5

kb.

19. GIG46

The gene of the present invention is a human cancer suppressor gene 1 (GIG46)

having a DNA sequence of SEQ ID NO: 73, which was deposited with Accession No.

AY692464 into the GenBank database of U.S. National Institutes of Health (NIH)

(Scheduled Release Date: May 1, 2006), and a DNA sequence of the deposited gene is

similar to that of the Homo sapiens actin, alpha 2, smooth muscle, aorta (ACTA2) gene

deposited with Accession No. NM_001613 into the database. Contrary to its functions

as reported previously, it was however found from this study result that the GIG46 gene

was closely related to various human carcinogenesis. From the study result, it was

found that the GIG8 tumor suppressor gene was very rarely expressed in various human

tumors including the uterine cancer, while its expression was significantly increased in

various normal tissues.

The DNA sequence of SEQ ID NO: 73 has one open reading frame (ORP)

corresponding to base positions from 393 to 1,526 of the DNA sequence (base positions

from 1,524 to 1,526 represent a stop codon).

A protein expressed from the gene of the present invention consists of 377

amino acid residues, and has an amino acid sequence of SEQ ID NO: 74 and a molecular weight of approximately 42 kDa.

according to the conventional methods on the basis of the information on the DNA sequence set forth in SEQ ID NO: 73. As another example, a 255-bp cDNA fragment

(corresponding to base positions from 1,323 to 1,577), which is not expressed in the

cancer tissue or the cancer cell line but differentially expressed in the normal tissue, may

be obtained by carrying out a reverse transcription-polymerase chain reaction (RT-PCR)

on the total RNA extracted from a normal tissue, and a cancer tissue or a cancer cell line

using a random primer H-AP 16 of SEQ ID NO: 75 (S'-AAGCTTTAGAGCG-S¹) and an

anchored oligo-dT primer of SEQ ID NO: 76 (5'-AAGCTTTTTTTTTTTA-S'), and the

resultant fragment, which is used as the probe, may be plaque-hybridized with a cDNA library to obtain a full-length cDNA clone.

It is regarded that the gene of the present invention is overexpressed in the

normal tissues, preferably the uterus, the brain, the heart, the skeletal muscles, the large

intestine, the thymus, the spleen, the kidney, the liver, the small intestine, the placenta,

the lungs and the peripheral blood leukocyte to suppress the carcinogenesis. The gene

of the present invention is mainly overexpressed in these tissues as an mRNA transcript having a size of approximately 1.5 kb, and an mRNA transcript having a size of

approximately 2.0 kb is also expressed in addition to the 1.5-kb mRNA transcript.

20. PIG33 The gene of the present invention is a human cancer suppressor gene (PIG33) having a DNA sequence of SEQ ID NO: 77, which was deposited with Accession No. AY513278 into the GenBank database of U.S. National Institutes of Health (NIH) (Scheduled Release Date: December 31, 2005), and a DNA sequence of the deposited gene is similar to that of the Homo sapiens SPARC-like 1 (mast9, hevin) gene deposited

with Accession No. BC033721 into the database. From this study result, it was

however found that the PIG35 tumor suppressor gene was rarely expressed in various

human tumors including the liver cancer, while its expression was significantly increased in the normal liver tissue.

The DNA sequence of SEQ ID NO: 77 has one open reading frame (ORF)

corresponding to base positions from 81 to 2,075 of the DNA sequence (base positions

from 2,073 to 2,075 represent a stop codon).

A protein expressed from the gene of the present invention consists of 664

amino acid residues, and has an amino acid sequence of SEQ ID NO: 78 and a

molecular weight of approximately 75 kDa.

The gene and the protein of the present invention may be separated from human

sequence set forth in SEQ ID NO: 77. As another example, a 256-bp cDNA fragment,

transcription-polymerase chain reaction (RT-PCR) on the total RNA extracted from a normal tissue, and a cancer tissue or a cancer cell line using a random primer H-AP2 of SEQ ID NO: 79 (5'-AAGCTTCGACTGT-S') and an anchored oligo-dT primer of SEQ

ID NO: 80 (5'-AAGCTTTTTTTTTTTA-S'), and the resultant fragment, which is used as

The present inventors inserted the full-length PIG33 cDNA into the expression

DH5 α / PIG33/pBAD/Thio-Topo.

It is regarded that the gene of the present invention is overexpressed in the

normal tissues, preferably the liver, the brain, the heart, the muscles, the large intestine, the thymus, the spleen, the kidney, the small intestine, the placenta and the lungs to suppress the carcinogenesis. Also, it is regarded that its gene expression is suppressed

in the leukemia, the uterine cancer, the colon cancer, the lung cancer and the skin cancer

to induce the carcinogenesis. The gene of the present invention is mainly

overexpressed in these tissues as an mRNA transcript having a size of approximately 3.0

kb.

21. PIG35 The gene of the present invention is a human cancer suppressor gene (PIG35) having a DNA sequence of S EQ ID NO: 81, which was deposited with Accession No.

AY513280 into the GenBank database of U.S. National Institutes of Health (NIH)

gene is similar to that of the Homo sapiens serine (or cysteine) proteinase inhibitor

deposited with Accession No. NM 002615 into the database. From this study result, it was however found that the PIG35 tumor suppressor gene was rarely expressed in various human tumors including the breast cancer, while its expression was significantly

increased in the normal liver tissue. The DNA sequence of SEQ ID NO: 81 has one open reading frame (ORF)

corresponding to base positions from 50 to 1,306 of the DNA sequence (base positions

from 1,304 to 1,306 represent a stop codon).

A protein expressed from the gene of the present invention consists of 418 amino acid residues, and has an amino acid sequence of SEQ ID NO: 82 and a molecular weight of approximately 46 kDa.

The gene and the protein of the present invention may be separated from human

sequence set forth in SEQ ID NO: 81. As another example, a 312-bp cDNA fragment,

normal tissue, and a cancer tissue or a cancer cell line using a random primer H-AP9 of

SEQ ID NO: 83 (5'-AAGCTTCATTCCG-S¹) and an anchored oligo-dT primer of SEQ ID NO: 84 (5'-AAGCTTTTTTTTTTTC-S'), and the resultant fragment, which is used as

clone. The present inventors inserted the full-length PIG35 cDNA into the expression

DH5 α /PIG35/pBAD/Thio-Topo. It is regarded that the gene of the present invention is overexpressed in the

normal tissues, preferably the liver, the heart, the muscles, the brain, the small intestine,

the lungs and the placenta to suppress the carcinogenesis. Also, it is regarded that its

gene expression is suppressed in the leukemia, the uterine cancer, the malignant

lymphoma, the colon cancer, the lung cancer and the skin cancer to induce the

tissues as an mRNA transcript having a size of approximately 1.7 kb.

22. PIG36

The gene of the present invention is a human cancer suppressor gene (PIG36)

having a DNA sequence of SEQ ID NO: 85, which was deposited with Accession No.

AY544129 into the GenBank database of U.S. National Institutes of Health (NIH)

(Scheduled Release Date: December 31, 2005), and a DNA sequence of the deposited gene is similar to that of the Homo sapiens ATP synthase, H+ transporting,

mitochondrial FO complex, subunit F6 (ATP5J) deposited with Accession No. NM_001685 into the database. From this study result, it was however found that the

PIG36 tumor suppressor gene was rarely expressed in various human tumors including

the liver cancer, while its expression was significantly increased in various normal

tissues.

The DNA sequence of SEQ ID NO: 85 has one open reading frame (ORF)

corresponding to base positions from 102 to 428 of the DNA sequence (base positions from 426 to 428 represent a stop codon).

A protein expressed from the gene of the present invention consists of 108

amino acid residues, and has an amino acid sequence of SEQ ID NO: 86 and a molecular weight of approximately 13 kDa.

sequence set forth in SEQ ID NO: 85. As another example, a 162-bp cDNA fragment,

normal tissue, and a cancer tissue or a cancer cell line using a random primer H-AP9 of SEQ ID NO: 87 (S'-AAGCTTCATTCCG-S¹) and an anchored oligo-dT primer of SEQ

ID NO: 88 (5'-AAGCTTTTTTTTTTTG-S'), and the resultant fragment, which is used as

clone. The present inventors inserted the full-length PIG36 cDNA into the expression

vector pBAD/Thio-Topo (Invitrogen, U. A.), and then transformed E. coli DH5 α with

DH5 α / PIG36/pBAD/Thio-Topo.

It is regarded that the gene of the present invention is overexpressed in the

normal tissues, preferably the liver, the heart, the muscles, kidney and the placenta to

suppress the carcinogenesis. Also, it is regarded that the gene of the present invention was suppressed in the leukemia, the uterine cancer, the malignant lymphoma, the colon cancer, the lung cancer and the skin cancer to induce the carcinogenesis. The gene of the present invention is mainly overexpressed in these tissues as an mRNA transcript having a size of approximately 1.0 kb.

23. MIG20

A DNA sequence of SEQ ID NO: 89 was deposited with Accession No.

AY871271 into the GenBank database of U.S. National Institutes of Health (NIH)

is similar to that of the Homo sapiens mRNA; cDNA DKFZp686C0390 gene deposited

with Accession No. BX537651 into the database. From this study result, it was

however found that the MIG20 tumor suppressor gene was not expressed at all in

various human tumors including the uterine cancer, while its expression was

significantly increased in various normal tissues.

The DNA sequence of SEQ ID NO: 89 has one open reading frame (ORF)

corresponding to base positions from 8 to 202 of the DNA sequence (base positions

from 200 to 202 represent a stop codon). Also, the DNA sequence of SEQ ID NO: 89 has another open reading frame corresponding to base positions from 233 to 442 of the

DNA sequence (base positions from 440 to 442 represent a stop codon).

A protein expressed from the gene of the present invention consists of 64 amino

acid residues, and has an amino acid sequence of SEQ ID NO: 90 and a molecular

weight of approximately 7 kDa.

The gene and the protein of the present invention may be separated from human tissues, or also be synthesized according to the known methods for synthesizing DNA or peptide. For example, the gene of the present invention may be screened and cloned according to the conventional methods on the basis of the information on the DNA

sequence set forth in SEQ ID NO: 89. As another example, a 311-bp cDNA fragment (corresponding to base positions from 2,067 to 2,377), which is not expressed in the

be obtained by carrying out a reverse transcription-polymerase chain reaction (RT-PCR) on the total RNA extracted from a normal tissue, and a cancer tissue or a cancer cell line

using a random primer H-AP32 of SEQ ID NO: 91 (5'-AAGCTTCCTGCAA-S¹) and an

anchored oligo-dT primer of SEQ ID NO: 92 (5¹-AAGCTTTTTTTTTTTA-3^l), and the resultant fragment, which is used as the probe, may be plaque-hybridized with a cDNA library to obtain a full-length cDNA clone.

It is regarded that the gene of the present invention is overexpressed in the

normal tissues, preferably the uterus, the heart, the skeletal muscle, the kidney and the

liver to suppress the carcinogenesis. The gene of the present invention is mainly

overexpressed in these tissues as an mRNA transcript having a size of approximately 4.4

kb, and mRNA transcripts having sizes of approximately 2.4 kb and 1.5 kb are also

expressed in addition to the 4.4-kb mRNA transcript.

24. PIG49

The gene of the present invention is a human cancer suppressor gene (GIG49) having a DNA sequence of SEQ ID NO: 93, which was deposited with Accession No.

AY524047 into the GenBank database of U.S. National Institutes of Health (NIH)

gene is similar to that of the Homo sapiens transducer of ERBB2, 1 (TOBl) gene deposited with Accession No. NM 005749 into the database. From this study result, it was however found that the GIG49 gene was closely related to various human carcinogenesis. From the study result, it was found that the GIG49 tumor suppressor gene was rarely expressed or not expressed in various human tumors including the

The DNA sequence of SEQ ID NO: 93 has one open reading frame (ORF)

corresponding to base positions from 11 to 1 ,048 of the DNA sequence (base positions

from 1,046 to 1,048 represent a stop codon). However, because of degeneracy of codons, or considering preference of codons for living organisms to express the gene,

the gene of the present invention may be variously modified in coding region without

changing an amino acid sequence of the protein expressed from the coding region, and

also be variously modified or changed in a region except the coding region within a

range that does not affect the gene expression. Such a modified gene is also included in the scope of the present invention. Accordingly, the present invention also includes

a polynucleotide having substantially the same DNA sequences as the above-mentioned gene; and fragments of the gene. The term "substantially the same polynucleotide"

means a DNA sequence having a sequence homology of at least 80 %, preferably at

least 90 %, and the most preferably at least 95 %.

A protein expressed from the gene of the present invention consists of 345

amino acid residues, and has an amino acid sequence of SEQ ID NO: 94 and a

molecular weight of approximately 38 kDa. Also, one or more amino acids may be substituted, added or deleted even in the amino acid sequence of the protein within a range that does not affect functions of the protein, and only some of the protein may be used depending on their usage. Such a modified amino acid sequence is also included in the scope of the present invention. Accordingly, the present invention also includes

a polypeptide having substantially the same amino acid sequence as the protein; and fragments thereof. The term "substantially the same polypeptide" means a polypeptide

having a sequence homology of at least 80 %, preferably at least 90 %, and the most preferably at least 95 %.

The gene and the protein of the present invention may be separated from human

according to the conventional methods on the basis of the information on the DNA sequence set forth in SEQ ID NO: 93. As another example, a 272-bp cDNA fragment,

SEQ ID NO: 95 (5'-AAGCTTCCACGTA-S') and an anchored oligo-dT primer of SEQ

ID NO: 96 (5^I-AAGCTTTTTTTTTTTA-3^I), and the resultant fragment, which is used as the probe, may be plaque-hybridized with a cDNA library to obtain a full-length cDNA

clone.

The gene prepared thus may be inserted into a vector for expression in the

microorganisms or animal cells, already known in the art, to obtain an expression vector, and then DNA of the gene may be replicated in a large quantity or its protein may be produced in a commercial quantity by introducing the expression vector into suitable host cells, for example Escherichia coli, a MCF-7 cell line, etc. Upon constructing the

expression vector, expression regulatory sequences such as a promoter and a terminator, autonomously replicating sequences, secretion signals, etc. may be suitably selected and combined depending on a kind of the host cell that produces the gene or the protein.

It is regarded that the gene of the present invention is overexpressed in the

normal tissues, preferably the breast, the muscles, the heart, the kidney, the liver and the

placenta to suppress the carcinogenesis. The gene of the present invention is mainly overexpressed in these tissues as an mRNA transcript having a size of approximately 2.4 kb. Also, an mRNA transcript having a size of approximately 1.5 kb is expressed in

addition to the 2.4-kb mRNA transcript.

25. PIG51

The gene of the present invention is a human cancer suppressor gene (PIG51) having a DNA sequence of SEQ ID NO: 97, which was deposited with Accession No.

AY542308 into the GenBank database of U.S. National Institutes of Health (NIH)

gene is similar to that of the Homo sapiens TBCl domain family, member 7 gene

deposited with Accession No. BC007054 into the database. A specific function of the

gene remains to be specified. From this study result, it was however found that the

PIG51 gene was closely related to various human carcinogenesis. From the study

result, it was found that the PIG51 tumor suppressor gene was rarely expressed or not expressed in various human tumors including the breast cancer, while its expression was

significantly increased in various normal tissues. The DNA sequence of SEQ ID NO: 97 has one open reading frame (ORF) corresponding to base positions from 59 to 802 of the DNA sequence (base positions

from 800 to 802 represent a stop codon).

A protein expressed from the gene of the present invention consists of 247 amino acid residues, and has an amino acid sequence of SEQ ID NO: 98 and a

molecular weight of approximately 28 kDa.

The gene and the protein of the present invention may be separated from human

sequence set forth in SEQ ID NO: 97. As another example, a 211-bp cDNA fragment, which is not expressed or rarely expressed in the cancer tissue or the cancer cell line but

normal tissue, and a cancer tissue or a cancer cell line using a random primer H-AP22 of

SEQ ID NO: 99 (5'-AAGCTTTTGATCC-S') and an anchored oligo-dT primer of SEQ

ID NO: 100 (5'-AAGCTTTTTTTTTTTG-3^l), and the resultant fragment, which is used

as the probe, may be plaque-hybridized with a cDNA library to obtain a full-length

cDNA clone.

It is regarded that the gene of the present invention is overexpressed in the

normal tissues, preferably the breast, the heart, the muscles, the thymus, the spleen, the

kidney, the liver, the placenta and the peripheral blood leukocyte to suppress the

tissues as an mRNA transcript having a size of approximately 1.0 kb. 26. MIG12

The gene of the present invention is a human cancer suppressor gene (MIG 12)

having a DNA sequence of SEQ ID NO: 101, which was deposited with Accession No. AY453400 into the GenBank database of U.S. National Institutes of Health (NIH)

(Scheduled Release Date: March 31, 2005), and a DNA sequence of the deposited gene

is similar to that of the Homo sapiens thymosin, beta 10 gene deposited with Accession

No. BCO 16731 into the database. From this study result, it was however found that the

MIG 12 tumor suppressor gene was rarely expressed in various human tumors including

the lung cancer, while its expression was significantly increased in various normal

tissues.

The DNA sequence of SEQ ID NO: 101 has one open reading frame (ORF)

corresponding to base positions from 29 to 163 of the DNA sequence (base positions

from 161 to 163 represent a stop codon).

A protein expressed from the gene of the present invention consists of 44 amino

acid residues, and has an amino acid sequence of SEQ ID NO: 102 and a molecular

weight of approximately 5 kDa.

sequence set forth in SEQ ID NO: 101. As another example, a 161-bp cDNA fragment, which is very rarely expressed in the cancer tissue or the cancer cell line but differentially expressed in the normal tissue, may be obtained by carrying out a reverse transcription-polymerase chain reaction (RT-PCR) on the total RNA extracted from a normal tissue, and a cancer tissue or a cancer cell line using a random primer H-AP 12 of SEQ ID NO: 103 (5'-AAGCTTGAGTGCT-S¹) and an anchored oligo-dT primer of SEQ ID NO: 104 (5'-AAGCTTTTTTTTTTTC-S'), and the resultant fragment, which is used

as the probe, may be plaque-hybridized with a cDNA library to obtain a full-length cDNA clone.

It is regarded that the gene of the present invention is overexpressed in the

normal tissues, preferably the lungs, the brain, the heart, the muscles, the large intestine,

the thymus, the spleen, the kidney, the liver, the small intestine, the placenta and the

peripheral blood leukocyte to suppress the carcinogenesis. The gene of the present invention is mainly overexpressed in these tissues as an mRNA transcript having a size

of approximately 0.5 kb, and mRNA transcripts having sizes of approximately 1.0 kb

and 0.8 kb are expressed in addition to the 0.5-kb mRNA transcript.

27. PIG37

The gene of the present invention is a human cancer suppressor gene (PIG37) having a DNA sequence of SEQ ID NO: 105, which was deposited with Accession No.

AY513281 into the GenBank database of U.S. National Institutes of Health (NIH)

gene is similar to that of the Homo sapiens inositol 1,4,5-trisphosphate 3-kinase B

(ITPKB) gene deposited with Accession No. NM_002221 into the database. From this

study result, it was however found that the PIG37 tumor suppressor gene was rarely expressed in various human tumors including the liver cancer, while its expression was

significantly increased in the normal liver tissue.

The DNA sequence of SEQ ID NO: 105 has one open reading frame (ORF) corresponding to base positions from 4 to 1,422 of the DNA sequence (base positions

from 1,420 to 1 ,422represent a stop codon). A protein expressed from the gene of the present invention consists of 472

amino acid residues, and has an amino acid sequence of SEQ ID NO: 106 and a

molecular weight of approximately 53 kDa.

The gene and the protein of the present invention may be separated from human

according to the conventional methods on the basis of the information on the DNA sequence set forth in SEQ ID NO: 105. As another example, a 263 -bp cDNA fragment,

normal tissue, and a cancer tissue or a cancer cell line using a random primer H-APlO of SEQ ID NO: 107 (5'-AAGCTTCCACGTA-S¹) and an anchored oligo-dT primer of SEQ

ID NO: 108 (5'-AAGCTTTTTTTTTTTG-S'), and the resultant fragment, which is used as the probe, may be plaque-hybridized with a cDNA library to obtain a full-length

cDNA clone. The present inventors inserted the full-length PIG33 cDNA into the

expression vector pBAD/Thio-Topo (Invitrogen, U.A.), and then transformed E. coli

DH5 α with the resultant expression vector to obtain a transformant, which was

designated E. coli DH5 α /PIGSS/pBAD/Thio-Topo.

It is regarded that the gene of the present invention is overexpressed in the normal tissues, preferably the liver, the brain, the heart, the muscles, the large intestine, the thymus, the spleen, the kidney, the small intestine, the placenta and the lungs to suppress the carcinogenesis. Also, it is regarded that the gene of the present invention was suppressed in the leukemia, the uterine cancer, the colon cancer, the lung cancer

and the skin cancer to induce the carcinogenesis. The gene of the present invention is mainly overexpressed in these tissues as an mRNA transcript having a size of

approximately 7.0 kb.

28. GIG44

The gene of the present invention is a human cancer suppressor gene 44 (GIG44)

having a DNA sequence of SEQ ID NO: 109, which was deposited with Accession No.

AY971350 into the GenBank database of U.S. National Institutes of Health (NIH)

(Scheduled Release Date: May 31, 2006), and a DNA sequence of the deposited gene is similar to that of the Homo sapiens putative insulin-like growth factor II associated

protein gene deposited with Accession No. BC042127 into the database. From this

study result, it was however found that the GIG44 gene was closely related to various

human carcinogenesis. From the study result, it was found that the GIG44 tumor

suppressor gene was rarely expressed or not expressed in various human tumors

including the breast cancer, while its expression was significantly increased in various

normal tissues.

The DNA sequence of SEQ ID NO: 109 has one open reading frame (ORF)

corresponding to base positions from 59 to 400 of the DNA sequence (base positions

from 398 to 400 represent a stop codon).

A protein expressed from the gene of the present invention consists of 113 amino acid residues, and has an amino acid sequence of SEQ ID NO: 110 and a molecular weight of approximately 12 kDa.

sequence set forth in SEQ ID NO: 109. As another example, a 221-bp cDNA fragment,

SEQ ID NO: 111 (5'-AAGCTTGAGTGCT-S¹) and an anchored oligo-dT primer of SEQ

ID NO: 112 (5¹-AAGCTTTTTTTTTTTG-3^l), and the resultant fragment, which is used as the probe, may be plaque-hybridized with a cDNA library to obtain a full-length

cDNA clone.

It is regarded that the gene of the present invention is overexpressed in the normal tissues, preferably the breast, the heart, the kidney, the liver, the placenta and the

overexpressed in these tissues as an mRNA transcript having a size of approximately 1.0

kb.

29. GIG31

The gene of the present invention is a human cancer suppressor gene 31 (GIG31)

having a DNA sequence of SEQ ID NO: 113, which was deposited with Accession No. AY971351 into the GenBank database of U.S. National Institutes of Health (NIH)

(Scheduled Release Date: May 31, 2006), and a DNA sequence of the deposited gene is

similar to that of the Homo sapiens regulator of G-protein signalling 2 gene deposited with Accession No. NM 002923 into the database. From this study result, it was

however found that the GIG31 gene was closely related to various human

carcinogenesis. From the study result, it was found that the GIG31 tumor suppressor

The DNA sequence of SEQ ID NO: 113 has one open reading frame (ORF)

corresponding to base positions from 14 to 649 of the DNA sequence (base positions from 647 to 649 represent a stop codon).

A protein expressed from the gene of the present invention consists of 211 amino acid residues, and has an amino acid sequence of SEQ ID NO: 114 and a

molecular weight of approximately 24 kDa.

The gene and the protein of the present invention may be separated from human

according to the conventional methods on the basis of the information on the DNA sequence set forth in SEQ ID NO: 113. As another example, a 223-bp cDNA fragment, which is not expressed or rarely expressed in the cancer tissue or the cancer cell line but

normal tissue, and a cancer tissue or a cancer cell line using a random primer H-AP4 of SEQ ID NO: 115 (5'-AAGCTTCTCAACG-S¹) and an anchored oligo-dT primer of SEQ ID NO: 116 (5'-AAGCTTTTTTTTTTT A-3¹), and the resultant fragment, which is used

It is regarded that the gene of the present invention is overexpressed in the

normal tissues, preferably the breast, the heart, the large intestine, the spleen, the small

intestine, the placenta, the lungs and the peripheral blood leukocyte to suppress the

tissues as an mRNA transcript having a size of approximately 1.4 kb.

Meanwhile, because of degeneracy of codons, or considering preference of codons for living organisms to express the genes, the genes of the present invention may

be variously modified in coding regions without changing an amino acid sequence of the

protein expressed from the coding region, and also be variously modified or changed in

a region except the coding region within a range that does not affect the gene expression.

Such a modified gene is also included in the scope of the present invention.

Accordingly, the present invention also includes polynucleotides having substantially

the same DNA sequences as the above-mentioned genes; and fragments of the genes.

The term "substantially the same polynucleotide" means a DNA sequence having a

sequence homology of at least 80 %, preferably at least 90 %, and the most preferably at

least 95 %.

Also, one or more amino acids may be substituted, added or deleted even in the amino acid sequences of the proteins of the present invention within a range that does

not affect functions of the proteins, and only some of the proteins may be used depending on their usage. Such a modified amino acid sequence is also included in the scope of the present invention. Accordingly, the present invention also includes

polypeptides having substantially the same amino acid sequences as the proteins; and fragments thereof. The term "substantially the same polypeptide" means a polypeptide

having sequence homology of at least 80 %, preferably at least 90 %, and the most preferably at least 95 %.

In some embodiments, the genes of the present invention prepared thus may be also inserted into a vector for expression in the microorganisms or animal cells, already

known in the art, to obtain expression vectors, and then DNA of the genes may be

replicated in a large quantity or its protein may be produced in a commercial quantity by

introducing the expression vectors into suitable host cells, for example Escherichia coli, a MCF-7 cell line, etc. Upon constructing the expression vectors, expression regulatory sequences such as a promoter and a terminator, autonomously replicating

sequences, secretion signals, etc. may be suitably selected and combined depending on

kinds of the host cells that produce the genes or the proteins.

Especially, the genes of the present invention are differentially expressed only in

the normal tissues. For example, their gene expressions are slightly detected or not

detected in the cancer tissues and the cancer cells such as the breast cancer tissue, the

breast cancer cell line MCF-7, the leukemia cell, the leukemia cell line K562, the liver cancer tissue, the liver cancer cell line HepG2, the cervical cancer tissue, the cervical cancer cell line, the lung cancer tissue, the metastatic lung cancer tissue and the lung

cancer cell lines (A549 and NCI-H358), but differentially increased only in the normal

uterine tissues.

The cancer cell lines introduced with the genes of the present invention showed a high mortality, and therefore the genes of the present invention may be effectively used for treatment and prevention of the cancer. Hereinafter, the present invention will be described in detail referring to

preferred examples. Therefore, the description proposed herein is just a preferable

example for the purpose of illustrations only, not intended to limit the scope of the invention.

Reference Example: Separation of Total RNA

The total RNA samples were separated from fresh tissues or cultured cells using

the RNeasy total RNA kit (Qiagen Inc., Germany), and the contaminated DNA was then

removed from the RNA samples using the message clean kit (GenHunter Corp., MA,

U.S.).

Example 1: Separation of Total RNA and Differential Display of mRNA

A differential expression pattern of the gene was investigated in a normal breast tissue, a primary breast cancer tissue and a breast cancer cell line, as follows.

A normal breast tissue sample was obtained from a breast cancer patient during

mastectomy, and a primary breast cancer tissue sample was obtained during radical

mastectomy from a breast cancer patient who has not been subject to the radiation therapy and/or anticancer chemotherapy before the surgical treatment. MCF-7

(American Type Culture Collection; ATCC Number HTB-22) was used as the human

breast cancer cell line. The total RNAs were separated from these tissues and cells in the same manner as described in the reference example.

In order to conduct the mRNA differential display of the GIGl 5, a bone marrow tissue was also obtained from a normal person, and a primary leukemic bone marrow

tissue was obtained from a leukemia patient who has not been previously subject to the anticancer chemotherapy and/or radiation therapy during the bone marrow biopsy.

K-562 (American Type Cell Collection; ATCC Number CCL-243) was used as the

human chronic myelogenous leukemia cell line in the differential display method. The

total RNAs were separated from these tissues and cells in the same manner as described

in the reference example.

Meanwhile, a differential expression pattern of the gene was investigated in a normal liver tissue, a primary liver cancer tissue and a liver cancer cell line in the case of the liver cancer-related genes, as follows.

A normal liver tissue sample and a liver cancer tissue sample were obtained

from a liver cancer patient during the tissue biopsy, and the liver cancer cell line HepG2

(American Type Culture Collection; ATCC Number HB-8065) was used as the human liver cancer cell line. The total RNAs were separated from these tissues and cells in

the same manner as described in the reference example.

Also, a differential expression pattern of the gene of interest was measured in a normal exocervical tissue, a primary cervical cancer tissue and a cervical cancer cell line,

as follows. A normal exocervical tissue sample was obtained from a patient suffering

from a uterine myoma during hysterectomy, and a primary cervical tumor tissue sample

and a metastatic iliac lymph node tumor tissue sample were obtained during radical hysterectomy from a patient who has not been subject to the radiation therapy and/or

anticancer chemotherapy before surgical treatment. CUMC-6 (Kim, J. W. et ah, Gynecol. Oncol. 62: 230-240, 1996) was used as the human cervical cancer cell line.

The total RNA samples were separated from these tissues and cells in the same manner

as described in the reference example. The total RNAs were separated from these tissues and cells in the same manner as described in the reference example.

Also, a differential expression pattern of the gene of interest was measured in a

normal lung tissue, a primary lung cancer tissue, a metastatic lung cancer tissue and a

lung cancer cell line, as follows. A normal lung tissue sample, a lung cancer tissue

sample and a metastatic lung cancer tissue sample were obtained from a lung cancer

patient during surgical operation. The lung cancer cell lines A549 (American Type

Culture Collection; ATCC Number CCL-185) and NCI-H358 (American Type Culture

Collection; ATCC Number CRL-5807) were used as the human lung cancer cell line.

The total RNAs were separated from these tissues and cells in the same manner as

described in the reference example.

A RT-PCR reaction was carried out using each of the total RNA samples

separated from the tissues and the cells according to the modified method as described

in the disclosure (Liang, P. and Pardee, A. B., Science, 257, 967-971 (1992); and Liang,

P. et al., Cancer Res., 52, 6966-6968 (1993)), as follows.

1-1. GIG8

0.2 μg of the total RNA was reverse-transcribed with an anchored oligo-dT

primer of SEQ ID NO: 4 using a kit (a RNAimage kit, GenHunter), and then a PCR

reaction was carried out in the presence of 0.5 mM [ α -³⁵S]-labeled dATP (1,200

Ci/mmol) using the same anchored oligo-dT primer and a random 5'-13-mer primer

H-AP33 (RNAimage primer set 5, GenHunter Corporation, U.S.) of SEQ ID NO: 3. The PCR reaction was conducted under the following conditions: the total 40

amplification cycles consisting of a denaturation step at 95 ^°C for 40 seconds, an

annealing step at 40 °C for 2 minutes and an extension step at 72 ^"C for 40 seconds, and followed by one extension step at 72 ^°C for 5 minutes. The amplified fragments

were electrophoresized in a 6 % polyacrylamide gel for DNA base sequence, and then

autoradiographed.

FIG. 1 shows a PCR result using a random 5'-13-mer primer H-AP33 of SEQ ID

NO: 3 and an anchored oligo-dT primer of SEQ ID NO: 4. In FIG. 1 , Lanes 1 , 2 and 3

represent a normal breast tissue; Lanes 4, 5 and 6 represent a breast cancer tissue; and

Lane 7 represents a breast cancer cell line MCF-7. As shown in FIG. 1, it was

confirmed that a 163-bp cDNA fragment (Base positions from 317 to 479 of the

full-length GIG8 gene sequence) was very rarely expressed in the breast cancer tissue

and the breast cancer cell line, but differentially expressed at an increased level only in

the normal lung tissue. This cDNA fragment was designated FC33.

A 163-bp band, FC33 fragment, was removed from the dried gell, boiled for 15

minutes to elute the cDNA, and a PCR reaction was then carried out under the same said

condition using the same primer set as described above to re-amplify the FC33 cDNA,

except that the [ α -³⁵S]-labeled dATP and the 20 μ M dNTP were not used herein.

The re-amplified cDNA fragment FC33 was cloned into an expression vector pGEM-T Easy using the TA cloning system (Promega), and then its DNA sequence was determined using the Sequenase Version 2.0 DNA Sequencing System (United States

Biochemical Co.). 1-2. GIGlO

0.2 jMg of the total RNA was reverse-transcribed with an anchored oligo-dT

primer of SEQ ID NO: 8 using a kit (a RNAimage kit, GenHunter), and then a PCR

reaction was carried out in the presence of 0.5 mM [ α -³⁵S]-labeled dATP (1,200 Ci/mmol) using the same anchored oligo-dT primer and a random 5'-13-mer primer

H-APlO (RNAimage primer set 2, GenHunter Corporation, U.S.) of SEQ ID NO: 7.

The PCR reaction was conducted under the following conditions: the total 40

annealing step at 40 ⁰C for 2 minutes and an extension step at 72 ^°C for 40 seconds,

and followed by one extension step at 72 ^°C for 5 minutes. The amplified fragments

autoradiographed.

FIG. 2 shows a PCR result using a random 5'-13-mer primer H-APlO of SEQ ID

NO: 7 and an anchored oligo-dT primer of SEQ ID NO: 8. In FIG. 2, Lanes 1 , 2 and 3

Lane 7 represents a breast cancer cell line MCF-7. As shown in FIG. 2, it was

confirmed that a 321-bp cDNA fragment (Base positions from 1,716 to 2,036 of the full-length GIGlO gene sequence) was very rarely expressed in the breast cancer tissue and the breast cancer cell line, but differentially expressed at an increased level only in

the normal breast tissue. This cDNA fragment was designated FC42.

A 321-bp band, FC42 fragment, was removed from the dried gell, boiled for 15

condition using the same primer set as described above to re-amplify the FC42 cDNA,

except that the [ α -³⁵S]-labeled dATP and the 2O u M dNTP were not used herein.

The re-amplified cDNA fragment FC42 was cloned into an expression vector pGEM-T Easy using the TA cloning system (Promega), and then its DNA sequence was determined using the Sequenase Version 2.0 DNA Sequencing System (United States Biochemical Co.).

1-3. GIGl 3

0.2 μg of the total RNA was reverse-transcribed with an anchored oligo-dT

primer of SEQ ID NO: 12 using a kit (a RNAimage kit, GenHunter), and then a PCR

reaction was carried out in the presence of 0.5 raM [ α -³⁵S]-labeled dATP (1,200

Ci/mmol) using the same anchored oligo-dT primer and a random 5'-13-mer primer

H-AP5 (RNAimage primer set 1, GenHunter Corporation, U.S.) of SEQ ID NO: 11.

The PCR reaction was conducted under the following conditions: the total 40

amplification cycles consisting of a denaturation step at 95 °C for 40 seconds, an

annealing step at 40 "C for 2 minutes and an extension step at 72 ^°C for 40 seconds,

autoradiographed.

FIG. 3 shows a PCR result using a random 5'-13-mer primer H-AP5 of SEQ ID

NO: 11 and an anchored oligo-dT primer of SEQ ID NO: 12. In FIG. 3, Lanes 1, 2 and

3 represent a normal breast tissue; Lanes 4, 5 and 6 represent a breast cancer tissue; and Lane 7 represents a breast cancer cell line MCF-7. As shown in FIG. 3, it was

confirmed that a 347-bp cDNA fragment (Base positions from 3,253 to 3,599 of the

full-length GIG 13 gene sequence) was very rarely expressed in the breast cancer tissue and the breast cancer cell line, but differentially expressed at an increased level only in

the normal breast tissue. This cDNA fragment was designated FC59.

A 347-bp band, FC59 fragment, was removed from the dried gell, boiled for 15 minutes to elute the cDNA, and a PCR reaction was then carried out under the same said condition using the same primer set as described above to re-amplify the FC59 cDNA,

The re-amplified cDNA fragment FC59 was cloned into an expression vector pGEM-T

Easy using the TA cloning system (Promega), and then its DNA sequence was

determined using the Sequenase Version 2.0 DNA Sequencing System (United States Biochemical Co.).

1-4. GIGl 5

In order to conduct the mRNA differential display, a bone marrow tissue was

also obtained from a normal person, and a primary leukemic bone marrow tissue was

obtained from a leukemia patient who has not been previously subject to the anticancer

chemotherapy and/or radiation therapy during the bone marrow biopsy. K-562

(American Type Cell Collection; ATCC Number CCL-243) was used as the human

chronic myelogenous leukemia cell line in the differential display method. The total

RNAs were separated from these tissues and cells in the same manner as described in

the reference example.

A RT-PCR reaction was carried out using each of the total RNA samples

P. et al., Cancer Res., 52, 6966-6968 (1993)), as follows. 0.2 μg of the total RNA

was reverse-transcribed with an anchored oligo-dT primer of SEQ ID NO: 16 using a kit (a RNAimage kit, GenHunter), and then a PCR reaction was carried out in the presence

of 0.5 ffiM [ α -³⁵S]-labeled dATP (1,200 Ci/mmol) using the same anchored oligo-dT

primer and a random 5'-13-mer primer H-AP2 (RNAimage primer set 5, GenHunter Corporation, U.S.) of SEQ ID NO: 15. The PCR reaction was conducted under the following conditions: the total 40 amplification cycles consisting of a denaturation step

at 95 ^°C for 40 seconds, an annealing step at 40 ^°C for 2 minutes and an extension

step at 72 ^°C for 40 seconds, and followed by one extension step at 72 ^°C for 5

minutes. The amplified fragments were electrophoresized in a 6 % polyacrylamide gel

for DNA base sequence, and then autoradiographed.

FIG. 4 shows a PCR result using a random 5'-13-mer primer H-AP2 of SEQ ID NO: 15 and an anchored oligo-dT primer of SEQ ID NO: 16. As shown in FIG. 4, it was confirmed that the gene is expressed at a different level in the normal bone marrow tissue and the leukemia cell and K-562 cell using the differential display (DD) method.

As seen in FIG. 4, a 133-bp cDNA fragment, GV2 (Base positions from 212 to 344 of

the full-length GIGl 5 gene sequence), was very rarely expressed in the leukemia tissue

and the K-562 cell, but highly expressed only in the normal bone marrow tissue. This

cDNA fragment was designated GV2. A 133-bp band, GV2 fragment, was removed

from the dried gell, boiled for 15 minutes to elute the cDNA, and a PCR reaction was then carried out under the same said condition using the same primer set as described

above to re-amplify the GV2 cDNA, except that the [ α -³⁵S]-labeled dATP and the 20

μ M dNTP were not used herein. The re-amplified cDNA fragment GV2 was cloned

into an expression vector pGEM-T Easy using the TA cloning system (Promega), and

then its DNA sequence was determined using the Sequenase Version 2.0 DNA Sequencing System (United States Biochemical Co.). 1-5. GIG16

A differential expression pattern of the gene was investigated in a normal liver tissue, a primary liver cancer tissue and a liver cancer cell line, as follows.

A normal liver tissue sample and a liver cancer tissue sample were obtained

(American Type Culture Collection; ATCC Number HB-8065) was used as the human

liver cancer cell line. The total RNAs were separated from these tissues and cells in

the same manner as described in the reference example.

A RT-PCR reaction was carried out using each of the total RNA samples separated from the tissues and the cells according to the modified method as described

P. et al, Cancer Res., 52, 6966-6968 (1993)), as follows. 0.2 βg of the total RNA

was reverse-transcribed with an anchored oligo-dT primer of SEQ ID NO: 20 using a kit

(a RNAimage kit, GenHunter), and then a PCR reaction was carried out in the presence

of 0.5 mM [ α -³⁵S]-labeled dATP (1,200 Ci/mmol) using the same anchored oligo-dT

primer and a random 5'-13-mer primer H-AP8 (RNAimage primer set 1, GenHunter

Corporation, U.S.) of SEQ ID NO: 19. The PCR reaction was conducted under the

following conditions: the total 40 amplification cycles consisting of a denaturation step

at 95 °C for 40 seconds, an annealing step at 40 ^°C for 2 minutes and an extension

step at 72 °C for 40 seconds, and followed by one extension step at 72 ^°C for 5

minutes. The amplified fragments were electrophoresized in a 6 % polyacrylamide gel for DNA base sequence, and then autoradiographed.

FIG. 5 shows a PCR result using a random 5'-13-mer primer H-AP8 of SEQ ID

NO: 19 and an anchored oligo-dT primer of SEQ ID NO: 20. In FIG. 5, Lanes 1 , 2 and

3 represent a normal liver tissue; Lanes 4, 5 and 6 represent a liver cancer tissue; and Lane 7 represents a liver cancer cell line HepG2. • As shown in FIG. 5, it was

confirmed that a 213-bp cDNA fragment (Base positions from 867 to 1,079 of the

full-length GIGl 6 gene sequence) was not expressed in the liver cancer tissue and the

liver cancer cell line, but differentially expressed only in the normal liver tissue. This

cDNA fragment was designated HP8.

A 213-bp band, HP 8 fragment, was removed from the dried gell, boiled for 15 minutes to elute the cDNA, and a PCR reaction was then carried out under the same said

condition using the same primer set as described above to re-amplify the HP8 cDNA,

The re-amplified cDNA fragment HP8 was cloned into an expression vector pGEM-T

Easy using the TA cloning system (Promega), and then its DNA sequence was

1-6. GIG24

A differential expression pattern of the gene was investigated in a normal liver

tissue, a primary liver cancer tissue and a liver cancer cell line, as follows.

A normal liver tissue sample and a liver cancer tissue sample were obtained

(American Type Culture Collection; ATCC Number HB-8065) was used as the human

liver cancer cell line. The total RNAs were separated from these tissues and cells in the same manner as described in the reference example.

A RT-PCR reaction was carried out using each of the total RNA samples separated from the tissues and the cells according to the modified method as described in the disclosure (Liang, P. and Pardee, A. B., Science, 257, 967-971 (1992); and Liang,

was reverse-transcribed with an anchored oligo-dT primer of SEQ ID NO: 24 using a kit

primer and a random 5'-13-mer primer H-AP7 (RNAimage primer set 1, GenHunter

Corporation, U.S.) of SEQ ID NO: 23. The PCR reaction was conducted under the

for DNA base sequence, and then autoradiographed.

FIG. 6 shows a PCR result using a random 5'-13-mer primer H-AP7 of SEQ ID

NO: 23 and an anchored oligo-dT primer of SEQ ID NO: 24. In FIG. 6, Lanes I₅ 2 and 3 represent a normal liver tissue; Lanes 4, 5 and 6 represent a liver cancer tissue; and

Lane 7 represents a liver cancer cell line HepG2. As shown in FIG. 6, it was

confirmed that a 221-bp cDNA fragment (Base positions from 1,057 to 1,277 of the

full-length GIG42 gene sequence) was not expressed in the liver cancer tissue and the

liver cancer cell line, but differentially expressed only in the normal liver tissue. This cDNA fragment was designated HP71.

A 221-bp band, HP71 fragment, was removed from the dried gell, boiled for 15

minutes to elute the cDNA, and a PCR reaction was then carried out under the same said condition using the same primer set as described above to re-amplify the HP71 cDNA, except that the [ α -³⁵S]-labeled dATP and the 20 μ M dNTP were not used herein.

The re-amplified cDNA fragment HP71 was cloned into an expression vector pGEM-T

Easy using the TA cloning system (Promega), and then its DNA sequence was

1-7. GIG26

0.2 βg of the total RNA was reverse-transcribed with an anchored oligo-dT

primer of SEQ ID NO: 28 using a kit (a RNAimage kit, GenHunter), and then a PCR

Ci/mmol) using the same anchored oligo-dT primer and a random 5'-13-mer primer H-API l (RNAimage primer set 1, GenHunter Corporation, U.S.) of SEQ ID NO: 27. The PCR reaction was conducted under the following conditions: the total 40

annealing step at 40 ^°C for 2 minutes and an extension step at 72 ^°C for 40 seconds,

were electrophoresized in a 6 % polyacrylamide gel for DNA base sequence, and then autoradiographed.

FIG. 7 shows a PCR result using a random 5'-13-mer primer H-APl 1 of SEQ ID

NO: 27 and an anchored oligo-dT primer of SEQ ID NO: 28. In FIG. 7, Lanes 1 , 2 and 3 represent a normal liver tissue; Lanes 4, 5 and 6 represent a liver cancer tissue; and

Lane 7 represents a liver cancer cell line HepG2. As shown in FIG. 7, it was

confirmed that a 204-bp cDNA fragment (Base positions from 1,036 to 1,239 of the full-length GIG26 gene sequence) was not expressed in the liver cancer tissue and the liver cancer cell line, but differentially expressed only in the normal liver tissue. This

cDNA fragment was designated HPl 15.

A 204-bp band, HPl 15 fragment, was removed from the dried gell, boiled for 15

condition using the same primer set as described above to re-amplify the HPl 15 cDNA,

The re-amplified cDNA fragment HPl 15 was cloned into an expression vector pGEM-T

Easy using the TA cloning system (Promega), and then its DNA sequence was

determined using the Sequenase Version 2.0 DNA Sequencing System (United States

Biochemical Co.).

1-8. GIG29

0.2 μg of the total RNA was reverse-transcribed with an anchored oligo-dT

primer of SEQ ID NO: 32 using a kit (a RNAimage kit, GenHunter), and then a PCR

Ci/mmol) using the same anchored oligo-dT primer and a random 5'-13-mer primer

H-AP3 (RNAimage primer set 1, GenHunter Corporation, U.S.) of SEQ ID NO: 31.

The PCR reaction was conducted under the following conditions: the total 40

amplification cycles consisting of a denaturation step at 95 "C for 40 seconds, an

annealing step at 40 °C for 2 minutes and an extension step at 72 ^°C for 40 seconds,

and followed by one extension step at 72 °C for 5 minutes. The amplified fragments

autoradiographed.

FIG. 8 shows a PCR result using a random 5'-13-mer primer H-AP3 of SEQ ID NO: 31 and an anchored oligo-dT primer of SEQ ID NO: 32. In FIG. 8, Lanes 1 , 2 and

3 represent a normal liver tissue; Lanes 4, 5 and 6 represent a liver cancer tissue; and

Lane 7 represents a liver cancer cell line HepG2. As shown in FIG. 8, it was

confirmed that a 277-bp cDNA fragment (Base positions from 823 to 1,099 of the

full-length GIG29 gene sequence) was not expressed in the liver cancer tissue and the

cDNA fragment was designated HP3.

A 277-bp band, HP3 fragment, was removed from the dried gell, boiled for 15

minutes to elute the cDNA, and a PCR reaction was then carried out under the same said condition using the same primer set as described above to re-amplify the HP3 cDNA,

The re-amplified cDNA fragment HP3 was cloned into an expression vector pGEM-T

Easy using the TA cloning system (Promega), and then its DNA sequence was

determined using the Sequenase Version 2.0 DNA Sequencing System (United States

Biochemical Co.). 1-9. GIG30

0.2 μg of the total RNA was reverse-transcribed with an anchored oligo-dT

primer of SEQ ID NO: 36 using a kit (a RNAimage kit, GenHunter), and then a PCR

Ci/mmol) using the same anchored oligo-dT primer and a random 5'-13-mer primer

H-AP4 (RNAimage primer set 1, GenHunter Corporation, U.S.) of SEQ ID NO: 35. The PCR reaction was conducted under the following conditions: the total 40

amplification cycles consisting of a denaturation step at 95 ^°C for 40 seconds, an annealing step at 40 ^°C for 2 minutes and an extension step at 72 ^°C for 40 seconds,

and followed by one extension step at 72 "C for 5 minutes. The amplified fragments

FIG. 9 shows a PCR result using a random 5'-13-mer primer H-AP4 of SEQ ID

NO: 35 and an anchored oligo-dT primer of SEQ ID NO: 36. In FIG. 9, Lanes 1 , 2 and

3 represent a normal breast tissue; Lanes 4, 5 and 6 represent a breast cancer tissue; and

Lane 7 represents a breast cancer cell line MCF-7. As shown in FIG. 9, it was

confirmed that a 278-bp cDNA fragment (Base positions from 1,462 to 1,739 of the

full-length GIG30 gene sequence) was very rarely expressed in the breast cancer tissue

and the breast cancer cell line, but differentially expressed at an increased level only in the normal breast tissue. This cDNA fragment was designated FC48.

A 278-bp band, FC48 fragment, was removed from the dried gell, boiled for 15

condition using the same primer set as described above to re-amplify the FC48 cDNA,

The re-amplified cDNA fragment FC48 was cloned into an expression vector pGEM-T

Easy using the TA cloning system (Promega), and then its DNA sequence was determined using the Sequenase Version 2.0 DNA Sequencing System (United States Biochemical Co.).

1-10. GIG32

0.2 βg of the total RNA was reverse-transcribed with an anchored oligo-dT

primer of SEQ ID NO: 40 using a kit (a RNAimage kit, GenHunter), and then a PCR reaction was carried out in the presence of 0.5 mM [ α -³⁵S]-labeled dATP (1,200

Ci/mmol) using the same anchored oligo-dT primer and a random 5'-13-mer primer

H-AP8 (RNAimage primer set 1, GenHunter Corporation, U.S.) of SEQ ID NO: 39.

The PCR reaction was conducted under the following conditions: the total 40

and followed by one extension step at 72 ⁰C for 5 minutes. The amplified fragments

autoradiographed. FIG. 10 shows a PCR result using a random 5'- 13-mer primer H-AP8 of SEQ ID

NO: 39 and an anchored oligo-dT primer of SEQ ID NO: 40. In FIG. 10, Lanes 1, 2

and 3 represent a normal breast tissue; Lanes 4, 5 and 6 represent a breast cancer tissue; and Lane 7 represents a breast cancer cell line MCF-7. As shown in FIG. 10, it was

confirmed that a 172-bp cDNA fragment tissue (Base positions from 428 to 599 of the

full-length GIG32 gene sequence) was very rarely expressed in the breast cancer tissue

the normal breast. This cDNA fragment was designated FC82.

A 172-bp band, FC82 fragment, was removed from the dried gell, boiled for 15 minutes to elute the cDNA, and a PCR reaction was then carried out under the same said condition using the same primer set as described above to re-amplify the FC82 cDNA,

The re-amplified cDNA fragment FC82 was cloned into an expression vector pGEM-T

Easy using the TA cloning system (Promega), and then its DNA sequence was determined using the Sequenase Version 2.0 DNA Sequencing System (United States

Biochemical Co.).

1-11. GIG33

0.2 μg of the total RNA was reverse-transcribed with an anchored oligo-dT

primer of SEQ ID NO: 44 using a kit (a RNAimage kit, GenHunter), and then a PCR

Ci/mmol) using the same anchored oligo-dT primer and a random 5'-13-mer primer

H-AP33 (RNAimage primer set 5, GenHunter Corporation, U.S.) of SEQ ID NO: 43. The PCR reaction was conducted under the following conditions: the total 40

annealing step at 40 ^°C for 2 minutes and an extension step at 72 ⁰C for 40 seconds,

autoradiographed. FIG. 11 shows a PCR result using a random 5'-13-mer primer H-AP33 of SEQ

ID NO: 43 and an anchored oligo-dT primer of SEQ ID NO: 44. In FIG. 11 , Lanes 1 , 2 and 3 represent a normal breast tissue; Lanes 4, 5 and 6 represent a breast cancer tissue;

and Lane 7 represents a breast cancer cell line MCF-7. As shown in FIG. 11, it was confirmed that a 182-bp cDNA fragment (Base positions from 216 to 397 of the full-length GIG33 gene sequence) was very rarely expressed in the breast cancer tissue

and the breast cancer cell line, but differentially expressed at an increased level only in the normal breast tissue. This cDNA fragment was designated FC86.

A 182-bp band, FC86 fragment, was removed from the dried gell, boiled for 15 minutes to elute the cDNA, and a PCR reaction was then carried out under the same said

condition using the same primer set as described above to re-amplify the FC86 cDNA,

The re-amplified cDNA fragment FC86 was cloned into an expression vector pGEM-T

Biochemical Co.). 1-12. GIG34

0.2 μg of the total RNA was reverse-transcribed with an anchored oligo-dT

primer of SEQ ID NO: 48 using a kit (a RNAimage kit, GenHunter), and then a PCR

Ci/mmol) using the same anchored oligo-dT primer and a random 5'-13-mer primer

H-AP35 (RNAimage primer set 5, GenHunter Corporation, U.S.) of SEQ ID NO: 47.

The PCR reaction was conducted under the following conditions: the total 40

autoradiographed. FIG. 12 shows a PCR result using a random 5'-13-mer primer H-AP35 of SEQ

ID NO: 47 and an anchored oligo-dT primer of SEQ ID NO: 48. In FIG. 12, Lanes 1 , 2

and 3 represent a normal breast tissue; Lanes 4, 5 and 6 represent a breast cancer tissue; and Lane 7 represents a breast cancer cell line MCF-7. As shown in FIG. 12, it was confirmed that a 205-bp cDNA fragment (Base positions from 343 to 547 of the

full-length GIG34 gene sequence) was very rarely expressed in the breast cancer tissue

the normal breast tissue. This cDNA fragment was designated FC35.

A 205-bp band, FC42 fragment, was removed from the dried gell, boiled for 15

condition using the same primer set as described above to re-amplify the FC35 cDNA,

The re-amplified cDNA fragment FC35 was cloned into an expression vector pGEM-T

Easy using the TA cloning system (Promega), and then its DNA sequence was

determined using the Sequenase Version 2.0 DNA Sequencing System (United States

Biochemical Co.).

1-13. GIG35

0.2 βg of the total RNA was reverse-transcribed with an anchored oligo-dT

primer of SEQ ID NO: 52 using a kit (a RNAimage kit, GenHunter), and then a PCR

Ci/mmol) using the same anchored oligo-dT primer and a random 5'-13-mer primer

H-AP3 (RNAimage primer set 1, GenHunter Corporation, U.S.) of SEQ ID NO: 51.

The PCR reaction was conducted under the following conditions: the total 40

FIG. 13 shows a PCR result using a random 5'-13-mer primer H-AP3 of SEQ ID

NO: 51 and an anchored oligo-dT primer of SEQ ID NO: 52. In FIG. 13, Lanes 1, 2

and 3 represent a normal breast tissue; Lanes 4, 5 and 6 represent a breast cancer tissue;

and Lane 7 represents a breast cancer cell line MCF-7. As shown in FIG. 13, it was

confirmed that a 212-bp cDNA fragment (Base positions from 1,108 to 1,319 of the

full-length GIG35 gene sequence) was very rarely expressed in the breast cancer tissue

the normal breast tissue. This cDNA fragment was designated FC38.

A 212-bp band, FC38 fragment, was removed from the dried gell, boiled for 15

condition using the same primer set as described above to re-amplify the FC38 cDNA,

The re-amplified cDNA fragment FC38 was cloned into an expression vector pGEM-T

Biochemical Co.).

1-14. GIG38

0.2 βg of the total RNA was reverse-transcribed with an anchored oligo-dT

primer of SEQ ID NO: 56 using a kit (a RNAimage kit, GenHunter), and then a PCR

Ci/mmol) using the same anchored oligo-dT primer and a random 5'-13-mer primer

H-AP 12 (RNAimage primer set 2, GenHunter Corporation, U.S.) of SEQ ID NO: 55. The PCR reaction was conducted under the following conditions: the total 40

FIG. 14 shows a PCR result using a random 5'-13-mer primer H-AP12 of SEQ

ID NO: 55 and an anchored oligo-dT primer of SEQ ID NO: 56. In FIG. 14, Lanes 1 , 2

and Lane 7 represents a breast cancer cell line MCF-7. As shown in FIG. 14, it was

confirmed that a 172-bp cDNA fragment (Base positions from 328 to 499 of the full-length GIG38 gene sequence) was very rarely expressed in the breast cancer tissue and the breast cancer cell line, but differentially expressed at an increased level only in

the normal breast tissue. This cDNA fragment was designated FC 122.

A 172-bp band, FC 122 fragment, was removed from the dried gell, boiled for 15 minutes to elute the cDNA, and a PCR reaction was then carried out under the same said condition using the same primer set as described above to re-amplify the FC 122 cDNA,

The re-amplified cDNA fragment FC 122 was cloned into an expression vector pGEM-T Easy using the TA cloning system (Promega), and then its DNA sequence was determined using the Sequenase Version 2.0 DNA Sequencing System (United States

Biochemical Co.).

1-15. GIG39 0.2 μg of the total RNA was reverse-transcribed with an anchored oligo-dT

primer of SEQ ID NO: 60 using a kit (a RNAimage kit, GenHunter), and then a PCR

Ci/mmol) using the same anchored oligo-dT primer and a random 5'-13-mer primer

H-AP12 (RNAimage primer set 2, GenHunter Corporation, U.S.) of SEQ ID NO: 59.

The PCR reaction was conducted under the following conditions: the total 40

autoradiographed.

FIG. 15 shows a PCR result using a random 5'-13-mer primer H-AP12 of SEQ

ID NO: 59 and an anchored oligo-dT primer of SEQ ID NO: 60. In FIG. 15, Lanes 1 , 2 and 3 represent a normal breast tissue; Lanes 4, 5 and 6 represent a breast cancer tissue; and Lane 7 represents a breast cancer cell line MCF-7. As shown in FIG. 15, it was

confirmed that a 327-bp cDNA fragment (Base positions from 2,533 to 2,859 of the full-length GIG39 gene sequence) was very rarely expressed in the breast cancer tissue

and the breast cancer cell line, but differentially expressed at an increased level only in the normal breast tissue. This cDNA fragment was designated FC 126. A 327-bp band, FC 126 fragment, was removed from the dried gell, boiled for 15

minutes to elute the cDNA, and a PCR reaction was then carried out under the same said condition using the same primer set as described above to re-amplify the FC 126 cDNA,

except that the [ α -³⁵S]-labeled dATP and the 20 μ M dNTP were not used herein. The re-amplified cDNA fragment FC 126 was cloned into an expression vector pGEM-T

Easy using the TA cloning system (Promega), and then its DNA sequence was

1-16. GIG40

0.2 μg of the total RNA was reverse-transcribed with an anchored oligo-dT

primer of SEQ ID NO: 64 using a kit (a RNAimage kit, GenHunter), and then a PCR

Ci/mmol) using the same anchored oligo-dT primer and a random 5'-13-mer primer

H-AP7 (RNAimage primer set 1, GenHunter Corporation, U.S.) of SEQ ID NO: 63.

The PCR reaction was conducted under the following conditions: the total 40

amplification cycles consisting of a denaturation step at 95 ⁰C for 40 seconds, an

autoradiographed.

FIG. 16 shows a PCR result using a random 5'-13-mer primer H-AP7 of SEQ ID

NO: 63 and an anchored oligo-dT primer of SEQ ID NO: 64. In FIG. 16, Lanes 1, 2

and 3 represent a normal liver tissue; Lanes 4, 5 and 6 represent a liver cancer tissue; and Lane 7 represents a liver cancer cell line HepG2. As shown in FIG. 16, it was

confirmed that a 275-bp cDNA fragment (Base positions from 3,112 to 3,386 of the

full-length GIG40 gene sequence) was rarely expressed in the liver cancer tissue and the liver cancer cell line, but differentially expressed only in the normal liver tissue. This cDNA fragment was designated HP79.

A 275-bp band, HP79 fragment, was removed from the dried gell, boiled for 15

condition using the same primer set as described above to re-amplify the HP79 cDNA,

The re-amplified cDNA fragment HP79 was cloned into an expression vector pGEM-T Easy using the TA cloning system (Promega), and then its DNA sequence was

determined using the Sequenase Version 2.0 DNA Sequencing System (United States

Biochemical Co.).

1-17. GIG42

0.2 βg of the total RNA was reverse-transcribed with an anchored oligo-dT

primer of SEQ ID NO: 68 using a kit (a RNAimage kit, GenHunter), and then a PCR

reaction was carried out in the presence of 0.5 mM [ α -³⁵S] -labeled dATP (1,200

Ci/mmol) using the same anchored oligo-dT primer and a random 5'-13-mer primer

H-AP8 (RNAimage primer set 1, GenHunter Corporation, U.S.) of SEQ ID NO: 67.

The PCR reaction was conducted under the following conditions: the total 40

autoradiographed.

FIG. 17 shows a PCR result using a random 5'-13-mer primer H-AP8 of SEQ ID

NO: 67 and an anchored oligo-dT primer of SEQ ID NO: 68. In FIG. 17, Lanes 1, 2 and 3 represent a normal liver tissue; Lanes 4, 5 and 6 represent a liver cancer tissue;

and Lane 7 represents a liver cancer cell line HepG2. As shown in FIG. 17, it was

confirmed that a 327-bp cDNA fragment (Base positions from 1,473 to 1,799 of the

cDNA fragment was designated HP85.

A 327-bp band, HP85 fragment, was removed from the dried gell, boiled for 15 minutes to elute the cDNA, and a PCR reaction was then carried out under the same said

condition using the same primer set as described above to re-amplify the HP85 cDNA,

The re-amplified cDNA fragment HP 85 was cloned into an expression vector pGEM-T

Easy using the TA cloning system (Promega), and then its DNA sequence was

determined using the Sequenase Version 2.0 DNA Sequencing System (United States

Biochemical Co.). 1-18. GIG43

0.2 βg of the total RNA was reverse-transcribed with an anchored oligo-dT

primer of SEQ ID NO: 72 using a kit (a RNAimage kit, GenHunter), and then a PCR

Ci/mmol) using the same anchored oligo-dT primer and a random 5'-13-mer primer H-APlO (RNAimage primer set 5, GenHunter Corporation, U.S.) of SEQ ID NO: 71. The PCR reaction was conducted under the following conditions: the total 40

amplification cycles consisting of a denaturation step at 95 ^"C for 40 seconds, an

annealing step at 40 °C for 2 minutes and an extension step at 72 ^°C for 40 seconds, and followed by one extension step at 72 °C for 5 minutes. The amplified fragments

FIG. 18 shows a PCR result using a random 5'-13-mer primer H-APlO of SEQ ID NO: 71 and an anchored oligo-dT primer of SEQ ID NO: 72. In FIG. 18, Lanes 1 , 2

and Lane 7 represents a breast cancer cell line MCF-7. As shown in FIG. 18, it was

confirmed that a 273-bp cDNA fragment (Base positions from 727 to 999 of the full-length GIG43 gene sequence) was very rarely expressed in the breast cancer tissue

the normal breast tissue. This cDNA fragment was designated FC 102.

A 273-bp band, FC 102 fragment, was removed from the dried gell, boiled for 15

condition using the same primer set as described above to re-amplify the FC 102 cDNA,

The re-amplified cDNA fragment FC 102 was cloned into an expression vector pGEM-T Easy using the TA cloning system (Promega), and then its DNA sequence was

determined using the Sequenase Version 2.0 DNA Sequencing System (United States

Biochemical Co.).

1-19. GIG46

A differential expression pattern of the gene of interest was measured in a normal exocervical tissue, a primary cervical cancer tissue and a cervical cancer cell line,

as follows. A normal exocervical tissue sample was obtained from a patient suffering from a uterine myoma during hysterectomy, and a primary cervical tumor tissue sample

and a metastatic iliac lymph node tumor tissue sample were obtained during radical

hysterectomy from a patient who has not been subject to the radiation therapy and/or

anticancer chemotherapy before surgical treatment. CUMC-6 (Kim, J. W. et al, Gynecol. Oncol. 62: 230-240, 1996) was used as the human cervical cancer cell line.

as described in the reference example. The total RNAs were separated from these

tissues and cells in the same manner as described in the reference example.

A RT-PCR reaction was carried out using each of the total RNA samples

was reverse-transcribed with an anchored oligo-dT primer of SEQ ID NO: 76 using a kit

primer and a random 5'-13-mer primer H-AP 16 (RNAimage primer set 5, GenHunter

Corporation, U.S.) of SEQ ID NO: 75. The PCR reaction was conducted under the following conditions: the total 40 amplification cycles consisting of a denaturation step

for DNA base sequence, and then autoradiographed.

FIG. 19 shows a PCR result using a random 5'-13-mer primer H-AP16 of SEQ ID NO: 75 and an anchored oligo-dT primer of SEQ ID NO: 76. In FIG. 19, Lane 1

represents a normal exocervical tissue; Lane 2 represents a cervical cancer tissue; Lane

3 represents a metastatic iliac lymph node tissue; and Lane 4 represents a cervical cancer

cell line CUMC-6. As shown in FIG. 19, it was confirmed that a 255-bp cDNA

fragment was not expressed in the cervical cancer tissue, the metastatic iliac lymph node

tissue and the cervical cancer cell line CUMC-6, but differentially expressed only in the

normal exocervical tissue. This cDNA fragment was designated CAl 61.

A 255-bp band, CA161 fragment, was removed from the dried gell, boiled for 15 minutes to elute the cDNA, and a PCR reaction was then carried out under the same said

condition using the same primer set as described above to re-amplify the CAl 61 cDNA,

except that the [ α -³⁵S] -labeled dATP and the 20 μ M dNTP were not used herein.

The re-amplified cDNA fragment CAl 61 was cloned into an expression vector

pGEM-T Easy using the TA cloning system (Promega), and then its DNA sequence was

determined using the Sequenase Version 2.0 DNA Sequencing System (United States

Biochemical Co.).

1-20. PIG33

0.2 βg of the total RNA was reverse-transcribed with an anchored oligo-dT

primer of SEQ ID NO: 80 using a kit (a RNAimage kit, GenHunter), and then a PCR

Ci/mmol) using the same anchored oligo-dT primer and a random 5'-13-mer primer H-AP2 (RNAimage primer set 1, GenHunter Corporation, U.S.) of SEQ ID NO: 79.

The PCR reaction was conducted under the following conditions: the total 40

amplification cycles consisting of a denaturation step at 95 ^°C for 40 seconds, an annealing step at 40 ^°C for 2 minutes and an extension step at 72 ⁰C for 40 seconds,

FIG. 20 shows a PCR result using a random 5'- 13-mer primer H-AP2 of SEQ ID

NO: 79 and an anchored oligo-dT primer of SEQ ID NO: 80. In FIG. 20, Lanes 1, 2 and 3 represent a normal liver tissue; Lanes 4, 5 and 6 represent a liver cancer tissue;

and Lane 7 represents a liver cancer cell line HepG2. As shown in FIG. 20, it was

confirmed that a 256-bp cDNA fragment (Base positions from 1,623 to 1,878 of the full-length PIG33 gene sequence) was not expressed or rarely expressed in the liver

cancer tissue and the liver cancer cell line, but differentially expressed only in the

normal liver tissue. This cDNA fragment was designated HP29.

A 256-bp band, HP29 fragment, was removed from the dried gell, boiled for 15

minutes to elute the cDNA, and a PCR reaction was then carried out under the same said condition using the same primer set as described above to re-amplify the HP29 cDNA,

The re-amplified cDNA fragment HP29 was cloned into an expression vector pGEM-T

Easy using the TA cloning system (Promega), and then its DNA sequence was

determined using the Sequenase Version 2.0 DNA Sequencing System (United States Biochemical Co.). 1-21. PIG35

0.2 βg of the total RNA was reverse-transcribed with an anchored oligo-dT

primer of SEQ ID NO: 84 using a kit (a RNAimage kit, GenHunter), and then a PCR reaction was carried out in the presence of 0.5 mM [ α -³⁵S]-labeled dATP (1,200

Ci/mmol) using the same anchored oligo-dT primer and a random 5'-13-mer primer

H-AP9 (RNAimage primer set 1, GenHunter Corporation, U.S.) of SEQ ID NO: 83.

The PCR reaction was conducted under the following conditions: the total 40

autoradiographed.

FIG. 21 shows a PCR result using a random 5'-13-mer primer H-AP9 of SEQ ID

NO: 83 and an anchored oligo-dT primer of SEQ ID NO: 84. In FIG. 21, Lanes 1, 2 and 3 represent a normal liver tissue; Lanes 4, 5 and 6 represent a liver cancer tissue;

and Lane 7 represents a liver cancer cell line HepG2. As shown in FIG. 21, it was

confirmed that a 312-bp cDNA fragment (Base positions from 966 to 1,277 of the

full-length PIG35 gene sequence) was not expressed or rarely expressed in the liver

normal liver tissue. This cDNA fragment was designated HP95.

A 312-bp band, HP95 fragment, was removed from the dried gell, boiled for 15

minutes to elute the cDNA, and a PCR reaction was then carried out under the same said condition using the same primer set as described above to re-amplify the HP95 cDNA,

The re-amplifϊed cDNA fragment HP95 was cloned into an expression vector pGEM-T

1-22. PIG36

0.2 βg of the total RNA was reverse-transcribed with an anchored oligo-dT

primer of SEQ ID NO: 88 using a kit (a RNAimage kit, GenHunter), and then a PCR

Ci/mmol) using the same anchored oligo-dT primer and a random 5'-13-mer primer

H-AP9 (RNAimage primer set 1, GenHunter Corporation, U.S.) of SEQ ID NO: 87.

The PCR reaction was conducted under the following conditions: the total 40

autoradiographed.

FIG. 22 shows a PCR result using a random 5'-l 3-mer primer H-AP9 of SEQ ID

NO: 87 and an anchored oligo-dT primer of SEQ ID NO: 88. In FIG. 22, Lanes 1, 2 and 3 represent a normal liver tissue; Lanes 4, 5 and 6 represent a liver cancer tissue; and Lane 7 represents a liver cancer cell line HepG2. As shown in FIG. 22, it was confirmed that a 162-bp cDNA fragment (Base positions from 238 to 399 of the full-length PIG36 gene sequence) was not expressed or rarely expressed in the liver

normal liver tissue. This cDNA fragment was designated HP96.

A 162-bp band, HP96 fragment, was removed from the dried gell, boiled for 15 minutes to elute the cDNA, and a PCR reaction was then carried out under the same said

condition using the same primer set as described above to re-amplify the HP96 cDNA,

The re-amplified cDNA fragment HP96 was cloned into an expression vector pGEM-T

Easy using the TA cloning system (Promega), and then its DNA sequence was

1-23. MIG20

0.2 βg of the total RNA was reverse-transcribed with an anchored oligo-dT

primer of SEQ ID NO: 92 using a kit (a RNAimage kit, GenHunter), and then a PCR

Ci/mmol) using the same anchored oligo-dT primer and a random 5'-13-mer primer

H-AP32 (RNAimage primer set 5, GenHunter Corporation, U.S.) of SEQ ID NO: 91.

The PCR reaction was conducted under the following conditions: the total 40

autoradiographed. FIG. 23 shows a PCR result using a random 5'-13-mer primer H-AP32 of SEQ

ID NO: 91 and an anchored oligo-dT primer of SEQ ID NO: 92. In FIG. 23, Lane 1 represents a normal exocervical tissue; Lane 2 represents a cervical cancer tissue; Lane

3 represents a metastatic iliac lymph node tissue; and Lane 4 represents a cervical cancer cell line CUMC-6. As shown in FIG. 23, it was confirmed that a 311-bp cDNA

fragment was not expressed in the cervical cancer tissue, the metastatic iliac lymph node tissue and the cervical cancer cell line CUMC-6, but differentially expressed only in the

normal exocervical tissue. This cDNA fragment was designated CA324.

A 311-bp band, C A324 fragment, was removed from the dried gell, boiled for 15

condition using the same primer set as described above to re-amplify the C A324 cDNA,

The re-amplified cDNA fragment C A324 was cloned into an expression vector

determined using the Sequenase Version 2.0 DNA Sequencing System (United States

Biochemical Co.).

1-24. PIG49

0.2 μg of the total RNA was reverse-transcribed with an anchored oligo-dT

primer of SEQ ID NO: 96 using a kit (a RNAimage kit, GenHunter), and then a PCR

Ci/mmol) using the same anchored oligo-dT primer and a random 5'-13-mer primer H-APlO (RNAimage primer set 2, GenHunter Corporation, U.S.) of SEQ ID NO: 95.

The PCR reaction was conducted under the following conditions: the total 40

FIG. 24 shows a PCR result using a random 5'-13-mer primer H-APlO of SEQ

ID NO: 95 and an anchored oligo-dT primer of SEQ ID NO: 96. In FIG. 24, Lanes 1 , 2

and Lane 7 represents a breast cancer cell line MCF-7. As shown in FIG. 24, it was

confirmed that a 272-bp cDNA fragment (Base positions from 767 to 1,038 of the full-length PIG49 gene sequence) was very rarely expressed in the breast cancer tissue

the normal breast tissue. This cDNA fragment was designated FClOl.

A 272-bp band, FClOl fragment, was removed from the dried gell, boiled for 15 minutes to elute the cDNA, and a PCR reaction was then carried out under the same said

condition using the same primer set as described above to re-amplify the FClOl cDNA,

The re-amplified cDNA fragment FClOl was cloned into an expression vector pGEM-T

Easy using the TA cloning system (Promega), and then its DNA sequence was

determined using the Sequenase Version 2.0 DNA Sequencing System (United States

Biochemical Co.).

1-25. PIG51

0.2 μg of the total RNA was reverse-transcribed with an anchored oligo-dT

primer of SEQ ID NO: 100 using a kit (a RNAimage kit, GenHunter), and then a PCR

Ci/mmol) using the same anchored oligo-dT primer and a random 5'-13-mer primer

H-AP22 (RNAimage primer set 1, GenHunter Corporation, U.S.) of SEQ ID NO: 99. The PCR reaction was conducted under the following conditions: the total 40

annealing step at 40 ^°C for 2 minutes and an extension step at 72 "C for 40 seconds,

autoradiographed.

FIG. 25 shows a PCR result using a random 5'-13-mer primer H-AP22 of SEQ

ID NO: 99 and an anchored oligo-dT primer of SEQ ID NO: 100. In FIG. 25, Lanes 1,

2 and 3 represent a normal breast tissue; Lanes 4, 5 and 6 represent a breast cancer

tissue; and Lane 7 represents a breast cancer cell line MCF-7. As shown in FIG. 25, it

was confirmed that a 211-bp cDNA fragment (Base positions from 519 to 729 of the

full-length PIG51 gene sequence) was very rarely expressed in the breast cancer tissue

the normal breast tissue. This cDNA fragment was designated FC22. A 211-bp band, FC22 fragment, was removed from the dried gell, boiled for 15

condition using the same primer set as described above to re-amplify the FC22 cDNA,

The re-amplified cDNA fragment FC22 was cloned into an expression vector pGEM-T Easy using the TA cloning system (Promega), and then its DNA sequence was determined using the Sequenase Version 2.0 DNA Sequencing System (United States

Biochemical Co.).

1-26. MIG12 0.2 μg of the total RNA was reverse-transcribed with an anchored oligo-dT

primer of SEQ ID NO: 104 using a kit (a RNAimage kit, GenHunter), and then a PCR

Ci/mmol) using the same anchored oligo-dT primer and a random 5'-13-mer primer

H-AP12 (RNAimage primer set 1, GenHunter Corporation, U.S.) of SEQ ID NO: 103.

The PCR reaction was conducted under the following conditions: the total 40

autoradiographed.

FIG. 26 shows a PCR result using a random 5'-13-mer primer H-AP12 of SEQ

ID NO: 103 and an anchored oligo-dT primer of SEQ ID NO: 104. In FIG. 26, Lane 1

represents a normal lung tissue; Lane 2 represents a lung cancer tissue; Lane 3

represents a metastatic lung cancer tissue; and Lane 4 represents a lung cancer cell line A549. As shown in FIG. 26, it was confirmed that a 161-bp cDNA fragment (Base positions from 35 to 195 of the full-length MIG12 gene sequence) was rarely expressed in the lung cancer tissue, the metastatic lung cancer tissue and the lung cancer cell line, but differentially expressed only in the normal lung tissue. This cDNA fragment was

designated L927.

A 161-bp band, L927 fragment, was removed from the dried gell, boiled for 15

minutes to elute the cDNA, and a PCR reaction was then carried out under the same said condition using the same primer set as described above to re-amplify the L927 cDNA, except that the [ α -³⁵S]-labeled dATP and the 20 μ M dNTP were not used herein.

The re-amplified cDNA fragment L927 was cloned into an expression vector pGEM-T

Easy using the TA cloning system (Promega), and then its DNA sequence was

determined using the Sequenase Version 2.0 DNA Sequencing System (United States

Biochemical Co.).

1-27. PIG37

0.2 μg of the total RNA was reverse-transcribed with an anchored oligo-dT

primer of SEQ ID NO: 108 using a kit (a RNAimage kit, GenHunter), and then a PCR

Ci/mmol) using the same anchored oligo-dT primer and a random 5'-13-mer primer

H-APlO (RNAimage primer set 1, GenHunter Corporation, U.S.) of SEQ ID NO: 107.

The PCR reaction was conducted under the following conditions: the total 40

autoradiographed.

FIG. 27 shows a PCR result using a random 5'-13-mer primer H-APlO of SEQ

ID NO: 107 and an anchored oligo-dT primer of SEQ ID NO: 108. In FIG. 27, Lanes 1, 2 and 3 represent a normal liver tissue; Lanes 4, 5 and 6 represent a liver cancer tissue; and Lane 7 represents a liver cancer cell line HepG2. As shown in FIG. 27, it was confirmed that a 263 -bp cDNA fragment (Base positions from 1,217 to 1,479 of the full-length PIG37 gene sequence) was not expressed or rarely expressed in the liver cancer tissue and the liver cancer cell line, but differentially expressed only in the

normal liver tissue. This cDNA fragment was designated HP 102.

A 263 -bp band, HP 102 fragment, was removed from the dried gell, boiled for 15 minutes to elute the cDNA, and a PCR reaction was then carried out under the same said

condition using the same primer set as described above to re-amplify the HP 102 cDNA,

The re-amplified cDNA fragment HP 102 was cloned into an expression vector pGEM-T Easy using the TA cloning system (Promega), and then its DNA sequence was

determined using the Sequenase Version 2.0 DNA Sequencing System (United States

Biochemical Co.).

1-28. GIG44

0.2 μg of the total RNA was reverse-transcribed with an anchored oligo-dT

primer of SEQ ID NO: 112 using a kit (a RNAimage kit, GenHunter), and then a PCR

Ci/mmol) using the same anchored oligo-dT primer and a random 5'-13-mer primer

H-AP12 (RNAimage primer set 5, GenHunter Corporation, U.S.) of SEQ ID NO: 111. The PCR reaction was conducted under the following conditions: the total 40

autoradiographed.

FIG. 28 shows a PCR result using a random 5'-13-mer primer H-AP12 of SEQ ID NO: 111 and an anchored oligo-dT primer of SEQ ID NO: 112. In FIG. 28, Lanes 1 ,

2 and 3 represent a normal breast tissue; Lanes 4, 5 and 6 represent a breast cancer tissue; and Lane 7 represents a breast cancer cell line MCF-7. As shown in FIG. 28, it

was confirmed that a 221-bp cDNA fragment (Base positions from 179 to 399 of the

full-length GIG44 gene sequence) was very rarely expressed in the breast cancer tissue

the normal breast tissue. This cDN A fragment was designated FC 123.

A 221-bp band, FC 123 fragment, was removed from the dried gell, boiled for 15

minutes to elute the cDNA, and a PCR reaction was then carried out under the same said condition using the same primer set as described above to re-amplify the FC 123 cDNA,

The re-amplified cDNA fragment FC 123 was cloned into an expression vector pGEM-T

Easy using the TA cloning system (Promega), and then its DNA sequence was

determined using the Sequenase Version 2.0 DNA Sequencing System (United States

Biochemical Co.).

1-29. GIG31

0.2 βg of the total RNA was reverse-transcribed with an anchored oligo-dT

primer of SEQ ID NO: 116 using a kit (a RNAimage kit, GenHunter), and then a PCR

Ci/mmol) using the same anchored oligo-dT primer and a random 5'-13-mer primer H-AP4 (RNAimage primer set 1, GenHunter Corporation, U.S.) of SEQ ID NO: 115.

The PCR reaction was conducted under the following conditions: the total 40

FIG. 29 shows a PCR result using a random 5'-l 3-mer primer H-AP4 of SEQ ID

NO: 115 and an anchored oligo-dT primer of SEQ ID NO: 116. In FIG. 29, Lanes 1 , 2

and Lane 7 represents a breast cancer cell line MCF-7. As shown in FIG. 29, it was confirmed that a 223 -bp cDNA fragment (Base positions from 445 to 667 of the

full-length GIG31 gene sequence) was very rarely expressed in the breast cancer tissue

the normal breast tissue. This cDNA fragment was designated FC47.

A 223 -bp band, FC47 fragment, was removed from the dried gell, boiled for 15

condition using the same primer set as described above to re-amplify the FC47 cDNA,

The re-amplified cDNA fragment FC47 was cloned into an expression vector pGEM-T

Easy using the TA cloning system (Promega), and then its DNA sequence was

Example 2: cDNA Library Screening

The cDNA fragments FC33; FC42; FC59; GV2; H-AP8; HP71; HPl 15; HP3; FC48; FC82; FC86; FC35; FC38; FC122; FC126; HP79; HP85; FC102; CA161; HP29; HP95; HP96; CA324; FClOl; FC22; L927; HP 102; FC 123 and FC47 obtained in

Example 1-1 were labeled according to the method of the disclosure (Feinberg, A.P. and

Vogelstein, B., Anal. Biochem., 132, 6-13 (1983)) to obtain ³²P-labeled probes FC33;

FC42; FC59; GV2; H-AP8; HP71; HP115; HP3; FC48; FC82; FC86; FC35; FC38;

FC 122; FC 126; HP79; HP85; FC 102; CAl 61; HP29; HP95; HP96; CA324; FClOl;

FC22; L927; P 102; FC 123 and FC47 cDNA, repectively, and the ³²P-labeled probes

were plaque-hybridized with bacteriophage λ gtl l human lung embryonic fibroblast

cDNA library (Miki, T. et al., Gene, 83, 137-146 (1989)) according to the method as

described in the disclosure (Sambrook, J. et al., Molecular Cloning: A Laboratory

manual, New York: Cold Spring Harbor Laboratory (1989)) to obtain full-length cDNA clones of the human cancer suppressor gene GIGs.

The full-length cDNA clones were sequenced, and therefore a DNA base

sequence result of the GIG8 was identical with SEQ ID NO: 1. The DNA sequence of the GIG8 has an open reading frame encoding 134 amino acid residues, and the amino

acid sequence derived from the open reading frame was identical with SEQ ID NO: 2.

The derived protein also had a molecular weight of approximately 15 kDa.

The transformed E. coli strain was incubated in LB broth, and then 1 mM

isopropy-1-β -D-thiogalactopyranoside (IPTG) was added to the culture broth, and

reacted at 37 °C for 3 hours to express the GIG8 gene. A protein sample was

obtained from the culture broth, and then SDS-PAGE was conducted with the protein sample according to the method as described in the disclosure (Sambrook, J. et al.,

Molecular Cloning: A Laboratory manual, New York: Cold Spring Harbor Laboratory

(1989)). FIG. 30 is a diagram showing an SDS-PAGE analysis of the GIG8 protein. In

FIG. 30, Lane 1 represents a protein sample before the IPTG induction, and Lane 2

represents a protein sample after the expression of the GIG8 gene is induced by IPTG.

As shown in FIG. 30, the expressed GIG8 protein has a molecular weight of

approximately 15 kDa, which corresponds to a molecular weight of a protein derived from its DNA sequence.

A DNA base sequence result of the GIGlO was identical with SEQ ID NO: 5.

The DNA sequence of the GIGlO has an open reading frame encoding 665 amino acid

residues, and the amino acid sequence derived from the open reading frame was

identical with SEQ ID NO: 6. The derived protein also had a molecular weight of approximately 73 kDa.

The transformed E. coli strain was incubated in LB broth, and then 1 mM

reacted at 37 ^°C for 3 hours to express the GIGlO gene. A protein sample was

obtained from the culture broth, and then SDS-PAGE was conducted with the protein

sample according to the method as described in the disclosure (Sambrook, J. et al.,

Molecular Cloning: A Laboratory manual, New York: Cold Spring Harbor Laboratory

(1989)).

FIG. 31 is a diagram showing an SDS-PAGE analysis of the GIGlO protein. In

FIG. 31, Lane 1 represents a protein sample before the IPTG induction, and Lane 2 represents a protein sample after the expression of the GIGlO gene is induced by IPTG.

As shown in FIG. 31, the expressed GIGlO protein has a molecular weight of

approximately 73 kDa, which corresponds to a molecular weight of a protein derived from its DNA sequence.

A DNA base sequence result of the GIGl 3 was identical with SEQ ID NO: 9.

The DNA sequence of the GIG 13 has an open reading frame encoding 1,201 amino acid

residues, and the amino acid sequence derived from the open reading frame was

identical with SEQ ID NO: 10. The derived protein also had a molecular weight of

approximately 132 kDa.

The transformed E. coli strain was incubated in LB broth, and then 1 mM

reacted at 37 ^°C for 3 hours to express the GIGl 3 gene. A protein sample was

sample according to the method as described in the disclosure (Sambrook, J. et al., Molecular Cloning: A Laboratory manual, New York: Cold Spring Harbor Laboratory

(1989)).

FIG. 32 is a diagram showing an SDS-PAGE analysis of the GIGl 3 protein. In

FIG. 32, Lane 1 represents a protein sample before the IPTG induction, and Lane 2

represents a protein sample after the expression of the GIGl 3 gene is induced by IPTG.

As shown in FIG. 32, the expressed GIGl 3 protein has a molecular weight of

approximately 132 kDa, which corresponds to a molecular weight of a protein derived from its DNA sequence. A DNA base sequence result of the GIG15 was identical with SEQ ID NO: 13.

The DNA sequence of the GIG 15 has an open reading frame encoding 106 amino acid residues, and the amino acid sequence derived from the open reading frame was identical with SEQ ID NO: 14. The derived protein also had a molecular weight of approximately 12 kDa.

The transformed E. coli strain was incubated in LB broth, and then 1 mM

reacted at 37 ^°C for 3 hours to express the GIG GIGl 5 gene. A protein sample was

Molecular Cloning: A Laboratory manual, New York: Cold Spring Harbor Laboratory

(1989)).

FIG. 33 is a diagram showing an SDS-PAGE analysis of the GIGl 5 protein. In FIG. 33, Lane 1 represents a protein sample before the IPTG induction, and Lane 2

represents a protein sample after the expression of the GIGl 5 gene is induced by IPTG.

As shown in FIG. 33, the expressed GIGl 5 protein has a molecular weight of

approximately 12 kDa, which corresponds to a molecular weight of a protein derived

from its DNA sequence. A DNA base sequence result of the GIG 16 was identical with SEQ ID NO: 17.

The DNA sequence of the GIG 16 has an open reading frame encoding 351 amino acid

residues, and the amino acid sequence derived from the open reading frame was identical with SEQ ID NO: 18. The derived protein also had a molecular weight of approximately 39 kDa. The resultant full-length GIG 16 cDNA was inserted into the prokaryotic expression vector pBAD/Thio-Topo (Invitrogen, U.A.), and then E. coli

DH5 α was transformed with the resultant expression vector to obtain a transformant,

which was designated E. coli DH5 α /GIGlό/pBAD/Thio-Topo.

The transformed E. coli strain was incubated in LB broth, and then 1 mM isopropy-1-β -D-thiogalactopyranoside (IPTG) was added to the culture broth, and

reacted at 37 ^°C for 3 hours to express the GIGl 6 gene. A protein sample was

Molecular Cloning: A Laboratory manual, New York: Cold Spring Harbor Laboratory

(1989)).

FIG. 34 is a diagram showing an SDS-PAGE analysis of the GIG 16 protein. In

FIG. 34, Lane 1 represents a protein sample before the IPTG induction, and Lane 2

represents a protein sample after the expression of the GIG 16 gene is induced by IPTG.

As shown in FIG. 34, the expressed GIGl 6 protein has a molecular weight of

approximately 39 kDa, which corresponds to a molecular weight of a protein derived from its DNA sequence.

A DNA base sequence result of the GIG24 was identical with SEQ ID NO: 21.

The DNA sequence of the GIG24 has an open reading frame encoding 423 amino acid

residues, and the amino acid sequence derived from the open reading frame was

identical with SEQ ID NO: 22. The derived protein also had a molecular weight of approximately 47 kDa. The resultant full-length GIG24 cDNA was inserted into the prokaryotic expression vector pBAD/Thio-Topo (Invitrogen, U.A.), and then E. coli

which was designated E. coli DH5 α /GIG24/pBAD/Thio-Topo.

The transformed E. coli strain was incubated in LB broth, and then 1 mM

reacted at 37 "C for 3 hours to express the GIG24 gene. A protein sample was obtained from the culture broth, and then SDS-PAGE was conducted with the protein

Molecular Cloning: A Laboratory manual, New York: Cold Spring Harbor Laboratory

(1989)).

FIG. 35 is a diagram showing an SDS-PAGE analysis of the GIG24 protein. In

FIG. 35, Lane 1 represents a protein sample before the IPTG induction, and Lane 2 represents a protein sample after the expression of the GIG24 gene is induced by IPTG.

As shown in FIG. 35, the expressed GIG24 protein has a molecular weight of

approximately 47 kDa, which corresponds to a molecular weight of a protein derived from its DNA sequence.

A DNA base sequence result of the GIG26 was identical with SEQ ID NO: 25.

The DNA sequence of the GIG26 has an open reading frame encoding 442 amino acid

residues, and the amino acid sequence derived from the open reading frame was identical with SEQ ID NO: 26. The derived protein also had a molecular weight of

approximately 50 kDa. The resultant full-length GIG26 cDNA was inserted into the

prokaryotic expression vector pBAD/Thio-Topo (Invitrogen, U.A.), and then E. coli

which was designated E. coli DH5 α /GIG26/pBAD/Thio-Topo.

The transformed E. coli strain was incubated in LB broth, and then 1 niM

reacted at 37 ^°C for 3 hours to express the GIG26 gene. A protein sample was

sample according to the method as described in the disclosure (Sambrook, J. et al., Molecular Cloning: A Laboratory manual, New York: Cold Spring Harbor Laboratory (1989)).

FIG. 36 is a diagram showing an SDS-PAGE analysis of the GIG26 protein. In FIG. 36, Lane 1 represents a protein sample before the IPTG induction, and Lane 2

represents a protein sample after the expression of the GIG26 gene is induced by IPTG.

As shown in FIG. 36, the expressed GIG26 protein has a molecular weight of

approximately 50 kDa, which corresponds to a molecular weight of a protein derived from its DNA sequence.

A DNA base sequence result of the GIG29 was identical with SEQ ID NO: 29.

The DNA sequence of the GIG29 has an open reading frame encoding 349 amino acid

residues, and the amino acid sequence derived from the open reading frame was

identical with SEQ ID NO: 30. The derived protein also had a molecular weight of

approximately 38 kDa. The resultant full-length GIG29 cDNA was inserted into the

which was designated E. coli DH5 α /GIG29/pBAD/Thio-Topo.

The transformed E. coli strain was incubated in LB broth, and then 1 mM

reacted at 37 ^°C for 3 hours to express the GIG29 gene. A protein sample was

obtained from the culture broth, and then SDS-PAGE was conducted with the protein sample according to the method as described in the disclosure (Sambrook, J. et al., Molecular Cloning: A Laboratory manual, New York: Cold Spring Harbor Laboratory

(1989)). FIG. 37 is a diagram showing an SDS-PAGE analysis of the GIG29 protein. In

FIG. 37, Lane 1 represents a protein sample before the IPTG induction, and Lane 2

represents a protein sample after the expression of the GIG29 gene is induced by IPTG.

As shown in FIG. 37, the expressed GIG29 protein has a molecular weight of approximately 38 kDa, which corresponds to a molecular weight of a protein derived

from its DNA sequence.

A DNA base sequence result of the GIG30 was identical with SEQ ID NO: 33.

The DNA sequence of the GIG30 has an open reading frame encoding 540 amino acid

residues, and the amino acid sequence derived from the open reading frame was

identical with SEQ ID NO: 34. The derived protein also had a molecular weight of approximately 61 kDa.

The transformed E. coli strain was incubated in LB broth, and then 1 mM

reacted at 37 ^°C for 3 hours to express the GIG30 gene. A protein sample was

Molecular Cloning: A Laboratory manual, New York: Cold Spring Harbor Laboratory

(1989)).

FIG. 38 is a diagram showing an SDS-PAGE analysis of the GIG30 protein. In

FIG. 38, Lane 1 represents a protein sample before the IPTG induction, and Lane 2 represents a protein sample after the expression of the GIG30 gene is induced by IPTG.

As shown in FIG. 38, the expressed GIG30 protein has a molecular weight of

approximately 61 kDa, which corresponds to a molecular weight of a protein derived from its DNA sequence.

A DNA base sequence result of the GIG32 was identical with SEQ ID NO: 37.

The DNA sequence of the GIG32 has an open reading frame encoding 178 amino acid

residues, and the amino acid sequence derived from the open reading frame was

identical with SEQ ID NO: 38. The derived protein also had a molecular weight of

approximately 20 kDa.

The transformed E. coli strain was incubated in LB broth, and then 1 mM

reacted at 37 ^°C for 3 hours to express the GIG32 gene. A protein sample was

Molecular Cloning: A Laboratory manual, New York: Cold Spring Harbor Laboratory

(1989)).

FIG. 39 is a diagram showing an SDS-PAGE analysis of the GIG32 protein. In

FIG. 39, Lane 1 represents a protein sample before the IPTG induction, and Lane 2

represents a protein sample after the expression of the GIG32 gene is induced by IPTG. As shown in FIG. 39, the expressed GIG32 protein has a molecular weight of

approximately 20 kDa, which corresponds to a molecular weight of a protein derived

from its DNA sequence. A DNA base sequence result of the GIG33 was identical with SEQ ID NO: 41.

The DNA sequence of the GIG33 has an open reading frame encoding 110 amino acid residues, and the amino acid sequence derived from the open reading frame was

identical with SEQ ID NO: 42. The derived protein also had a molecular weight of approximately 12 kDa.

The transformed E. coli strain was incubated in LB broth, and then 1 mM

reacted at 37 ^°C for 3 hours to express the GIG34 gene. A protein sample was

Molecular Cloning: A Laboratory manual, New York: Cold Spring Harbor Laboratory

(1989)).

FIG. 40 is a diagram showing an SDS-PAGE analysis of the GIG33 protein. In FIG. 40, Lane 1 represents a protein sample before the IPTG induction, and Lane 2

represents a protein sample after the expression of the GIG33 gene is induced by IPTG.

As shown in FIG. 40, the expressed GIG33 protein has a molecular weight of

from its DNA sequence. A DNA base sequence result of the GIG34 was identical with SEQ ID NO: 45.

The DNA sequence of the GIG34 has an open reading frame encoding 177 amino acid residues, and the amino acid sequence derived from the open reading frame was

identical with SEQ ID NO: 46. The derived protein also had a molecular weight of

approximately 20 kDa.

The transformed E. coli strain was incubated in LB broth, and then 1 mM

isopropy-1- β -D-thiogalactopyranoside (IPTG) was added to the culture broth, and

reacted at 37 ^°C for 3 hours to express the GIG34 gene. A protein sample was

Molecular Cloning: A Laboratory manual, New York: Cold Spring Harbor Laboratory

(1989)).

FIG. 41 is a diagram showing an SDS-PAGE analysis of the GIG34 protein. In

FIG. 41, Lane 1 represents a protein sample before the IPTG induction, and Lane 2

represents a protein sample after the expression of the GIG34 gene is induced by IPTG.

As shown in FIG. 41, the expressed GIG34 protein has a molecular weight of

from its DNA sequence.

A DNA base sequence result of the GIG35 was identical with SEQ ID NO: 49.

The DNA sequence of the GIG35 has an open reading frame encoding 437 amino acid residues, and the amino acid sequence derived from the open reading frame was

identical with SEQ ID NO: 50. The derived protein also had a molecular weight of

approximately 50 kDa.

The transformed E. coli strain was incubated in LB broth, and then 1 mM

reacted at 37 ^°C for 3 hours to express the GIG35 gene. A protein sample was

Molecular Cloning: A Laboratory manual, New York: Cold Spring Harbor Laboratory (1989)).

FIG. 42 is a diagram showing an SDS-PAGE analysis of the GIG35 protein. In

FIG. 42, Lane 1 represents a protein sample before the IPTG induction, and Lane 2 represents a protein sample after the expression of the GIG35 gene is induced by IPTG.

As shown in FIG. 42, the expressed GIG35 protein has a molecular weight of

approximately 50 kDa, which corresponds to a molecular weight of a protein derived

from its DNA sequence.

A DNA base sequence result of the GIG38 was identical with SEQ ID NO: 53.

The DNA sequence of the GIG38 has an open reading frame encoding 153 amino acid

residues, and the amino acid sequence derived from the open reading frame was

identical with SEQ ID NO: 54. The derived protein also had a molecular weight of approximately 17 kDa.

The transformed E. coli strain was incubated in LB broth, and then 1 mM

reacted at 37 ^°C for 3 hours to express the GIG38 gene. A protein sample was

(1989)).

FIG. 43 is a diagram showing an SDS-PAGE analysis of the GIG38 protein. In

FIG. 43, Lane 1 represents a protein sample before the IPTG induction, and Lane 2 represents a protein sample after the expression of the GIG38 gene is induced by IPTG. As shown in FIG. 43, the expressed GIG38 protein has a molecular weight of approximately 17 kDa, which corresponds to a molecular weight of a protein derived from its DNA sequence.

A DNA base sequence result of the GIG39 was identical with SEQ ID NO: 57. The DNA sequence of the GIG39 has an open reading frame encoding 928 amino acid

residues, and the amino acid sequence derived from the open reading frame was

identical with SEQ ID NO: 58. The derived protein also had a molecular weight of approximately 103 kDa.

The transformed E. coli strain was incubated in LB broth, and then 1 mM

reacted at 37 ^°C for 3 hours to express the GIG39 gene. A protein sample was

(1989)).

FIG. 44 is a diagram showing an SDS-PAGE analysis of the GIG39 protein. In

FIG. 44, Lane 1 represents a protein sample before the IPTG induction, and Lane 2

represents a protein sample after the expression of the GIG39 gene is induced by IPTG.

As shown in FIG. 44, the expressed GIG39 protein has a molecular weight of approximately 103 kDa, which corresponds to a molecular weight of a protein derived from its DNA sequence.

A DNA base sequence result of the GIG40 was identical with SEQ ID NO: 61.

The DNA sequence of the GIG40 has an open reading frame encoding 1,210 amino acid

residues, and the amino acid sequence derived from the open reading frame was identical with SEQ ID NO: 62. The derived protein also had a molecular weight of approximately 134 kDa. The resultant full-length GIG40 cDNA was inserted into the

prokaryotic expression vector pBAD/Thio-Topo (Invitrogen, U.A.), and then E. coli DH5 α was transformed with the resultant expression vector to obtain a transformant,

which was designated E. coli DH5 α /GIG40/pBAD/Thio-Topo.

The transformed E. coli strain was incubated in LB broth, and then 1 mM

reacted at 37 ^°C for 3 hours to express the GIG40 gene. A protein sample was

Molecular Cloning: A Laboratory manual, New York: Cold Spring Harbor Laboratory

(1989)).

FIG. 45 is a diagram showing an SDS-PAGE analysis of the GIG40 protein. In

FIG. 45, Lane 1 represents a protein sample before the IPTG induction, and Lane 2

represents a protein sample after the expression of the GIG40 gene is induced by IPTG.

As shown in FIG. 45, the expressed GIG40 protein has a molecular weight of

approximately 134 kDa, which corresponds to a molecular weight of a protein derived

from its DNA sequence.

A DNA base sequence result of the GIG42 was identical with SEQ ID NO: 65. The DNA sequence of the GIG42 has an open reading frame encoding 609 amino acid

residues, and the amino acid sequence derived from the open reading frame was

identical with SEQ ID NO: 66. The derived protein also had a molecular weight of approximately 69 kDa. The resultant full-length GIG42 cDNA was inserted into the

which was designated E. coli DH5 α /GIG42/pBAD/Thio-Topo. The transformed E. coli strain was incubated in LB broth, and then 1 mM

reacted at 37 ^°C for 3 hours to express the GIG42 gene. A protein sample was

FIG. 46 is a diagram showing an SDS-PAGE analysis of the GIG42 protein. In

FIG. 46, Lane 1 represents a protein sample before the IPTG induction, and Lane 2

represents a protein sample after the expression of the GIG42 gene is induced by IPTG.

As shown in FIG. 46, the expressed GIG42 protein has a molecular weight of

approximately 69 kDa, which corresponds to a molecular weight of a protein derived from its DNA sequence.

A DNA base sequence result of the GIG43 was identical with SEQ ID NO: 69.

The DNA sequence of the GIG43 has an open reading frame encoding 329 amino acid

residues, and the amino acid sequence derived from the open reading frame was

identical with SEQ ID NO: 70. The derived protein also had a molecular weight of

approximately 37 kDa.

The transformed E. coli strain was incubated in LB broth, and then 1 mM

reacted at 37 ^°C for 3 hours to express the GIG43 gene. A protein sample was

FIG. 47 is a diagram showing an SDS-PAGE analysis of the GIG43 protein. In

FIG. 47, Lane 1 represents a protein sample before the IPTG induction, and Lane 2

represents a protein sample after the expression of the GIG43 gene is induced by IPTG.

As shown in FIG. 47, the expressed GIG43 protein has a molecular weight of

approximately 37 kDa, which corresponds to a molecular weight of a protein derived from its DNA sequence.

A DNA base sequence result of the GIG46 was identical with SEQ ID NO: 73.

The DNA sequence of the GIG46 has an open reading frame encoding 377 amino acid

residues, and the amino acid sequence derived from the open reading frame was

identical with SEQ ID NO: 74. The derived protein also had a molecular weight of

approximately 42 kDa.

The resultant full-length GIG46 cDNA clone was inserted into a multi-cloning

site of the prokaryotic expression vector pBAD/thio-Topo (Invitrogen, U.S.) to obtain a

vector pBAD/thio-Topo/GIG46, and Escherichia coli ToplO (Invitrogen, U.S.) was then transformed with the resultant pBAD/thio-Topo/GIG46. The expression protein

HT-Thioredoxin is inserted upstream of the multi-cloning site of the vector

pBAD/thio-Topo. The transformed E. coli strain was incubated in LB broth with

shaking, and the resultant culture broth was diluted 1/100, and then incubated for 3 hours again. 0.5 mM L-arabinose (Sigma, U.S.) was added to the incubated culture broth to induce production of proteins. The E. coli cell in the culture broth was sonicated before and after the L-arabinose induction, and then 12 % sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) was conducted with the

sonicated homogenate. FIG. 48 is a diagram showing an expression pattern of proteins of the E. coli Top 10 strain transformed with the vector pBAD/thio-Topo/GIG46 using

the SDS-PAGE, wherein a band of a fusion protein having a molecular weight of

approximately 57 kDa was clearly observed after the L-arabinose induction. The

57-kDa fusion protein includes the approximately 15-kDa HT-thioredoxin protein

inserted into the vector pBAD/thio-Topo/GIG46 and the approximately 42-kDa GIG46 protein.

FIG. 48 is a diagram showing an SDS-PAGE analysis of the GIG46 protein. In FIG. 48, Lane 1 represents a protein sample before the L-arabinose induction, and Lane

2 represents a protein sample after the expression of the GIG46 gene is induced by

L-arabinose.

A DNA base sequence result of the PIG33 was identical with SEQ ID NO: 77.

The DNA sequence of the PIG33 has an open reading frame encoding 664 amino acid

residues, and the amino acid sequence derived from the open reading frame was identical with SEQ ID NO: 78. The derived protein also had a molecular weight of

approximately 75 kDa. The resultant full-length PIG33 cDNA was inserted into the prokaryotic expression vector pBAD/Thio-Topo (Invitrogen, U.A.), and then E. coli

which was designated E. coli DH5 α /PIG33/pBAD/Thio-Topo.

The transformed E. coli strain was incubated in LB broth, and then 1 mM

reacted at 37 ^°C for 3 hours to express the PIG33 gene. A protein sample was obtained from the culture broth, and then SDS-PAGE was conducted with the protein

FIG. 49 is a diagram showing an SDS-PAGE analysis of the PIG33 protein. In

FIG. 49, Lane 1 represents a protein sample before the IPTG induction, and Lane 2

represents a protein sample after the expression of the PIG33 gene is induced by IPTG.

As shown in FIG. 49, the expressed PIG33 protein has a molecular weight of

approximately 75 kDa, which corresponds to a molecular weight of a protein derived from its DNA sequence.

A DNA base sequence result of the PIG35 was identical with SEQ ID NO: 81.

The DNA sequence of the PIG35 has an open reading frame encoding 418 amino acid

residues, and the amino acid sequence derived from the open reading frame was

identical with SEQ ID NO: 82. The derived protein also had a molecular weight of

approximately 46 kDa. The resultant full-length PIG35 cDNA was inserted into the prokaryotic expression vector pBAD/Thio-Topo (Invitrogen, U.A.), and then E. coli

which was designated E. coli DH5 α /PIGSS/pBAD/Thio-Topo.

The transformed E. coli strain was incubated in LB broth, and then 1 mM

reacted at 37 ^°C for 3 hours to express the PIG35 gene. A protein sample was

obtained from the culture broth, and then SDS-PAGE was conducted with the protein sample according to the method as described in the disclosure (Sambrook, J. et al., Molecular Cloning: A Laboratory manual, New York: Cold Spring Harbor Laboratory (1989)).

FIG. 50 is a diagram showing an SDS-PAGE analysis of the PIG35 protein. In

FIG. 50, Lane 1 represents a protein sample before the IPTG induction, and Lane 2 represents a protein sample after the expression of the PIG35 gene is induced by IPTG.

As shown in FIG. 50, the expressed PIG35 protein has a molecular weight of

approximately 46 kDa, which corresponds to a molecular weight of a protein derived

from its DNA sequence.

A DNA base sequence result of the PIG 36 was identical with SEQ ID NO: 85. The DNA sequence of the PIG36 has an open reading frame encoding 108 amino acid

residues, and the amino acid sequence derived from the open reading frame was

identical with SEQ ID NO: 86. The derived protein also had a molecular weight of

approximately 13 kDa. The resultant full-length PIG36 cDNA was inserted into the

which was designated E. coli DH5 α /PIGSό/pBAD/Thio-Topo.

The transformed E. coli strain was incubated in LB broth, and then 1 mM

reacted at 37 ^°C for 3 hours to express the PIG36 gene. A protein sample was

(1989)). FIG. 51 is a diagram showing an SDS-PAGE analysis of the PIG36 protein. In

FIG. 51, Lane 1 represents a protein sample before the IPTG induction, and Lane 2

represents a protein sample after the expression of the PIG36 gene is induced by IPTG.

As shown in FIG. 51, the expressed PIG36 protein has a molecular weight of

approximately 13 kDa, which corresponds to a molecular weight of a protein derived from its DNA sequence.

A DNA base sequence result of the MIG20 was identical with SEQ ID NO: 89.

The DNA sequence of the MIG20 has an open reading frame encoding 64 amino acid

residues, and the amino acid sequence derived from the open reading frame was

identical with SEQ ID NO: 90. The derived protein also had a molecular weight of

approximately 7 kDa.

The resultant full-length MIG20 cDNA clone was inserted into a multi-cloning site of the prokaryotic expression vector pBAD/thio-Topo (Invitrogen, U.S.) to obtain a

vector pBAD/thio-Topo/MIG20, and Escherichia coli ToplO (Invitrogen, U.S.) was

then transformed with the resultant pBAD/thio-Topo/MIG20. The expression protein HT-Thioredoxin is inserted upstream of the multi-cloning site of the vector

pBAD/thio-Topo. The transformed E. coli strain was incubated in LB broth with

shaking, and the resultant culture broth was diluted 1/100, and then incubated for 3 hours again. 0.5 niM L-arabinose (Sigma, U.S.) was added to the incubated culture

broth to induce production of proteins. The E. coli cell in the culture broth was sonicated before and after the L-arabinose induction, and then 12 % sodium dodecyl

sulphate polyacrylamide gel electrophoresis (SDS-PAGE) was conducted with the

sonicated homogenate. FIG. 52 is a diagram showing an expression pattern of proteins of the E. coli Top 10 strain transformed with the vector pBAD/thio-Topo/MIG20 using

the SDS-PAGE, wherein a band of a fusion protein having a molecular weight of

approximately 22 kDa was clearly observed after the L-arabinose induction. The

22-kDa fusion protein includes the approximately 15 -kDa HT-thioredoxin protein

inserted into the vector pBAD/thio-Topo/MIG20 and the approximately 7-kDa MIG20

protein.

FIG. 52 is a diagram showing an SDS-PAGE analysis of the MIG20 protein. In

FIG. 52, Lane 1 represents a protein sample before the L-arabinose induction, and Lane

2 represents a protein sample after the expression of the MIG20 gene is induced by

L-arabinose.

A DNA base sequence result of the PIG49 was identical with SEQ ID NO: 93.

The DNA sequence of the PIG49 has an open reading frame encoding 345 amino acid

residues, and the amino acid sequence derived from the open reading frame was

identical with SEQ ID NO: 94. The derived protein also had a molecular weight of

approximately 38 kDa.

The transformed E. coli strain was incubated in LB broth, and then 1 mM

reacted at 37 °C for 3 hours to express the PIG49 gene. A protein sample was

(1989)).

FIG. 53 is a diagram showing an SDS-PAGE analysis of the PIG49 protein. In FIG. 53, Lane 1 represents a protein sample before the IPTG induction, and Lane 2

represents a protein sample after the expression of the PIG49 gene is induced by IPTG. As shown in FIG. 53, the expressed PIG49 protein has a molecular weight of

approximately 38 kDa, which corresponds to a molecular weight of a protein derived

from its DNA sequence.

A DNA base sequence result of the PIG51 was identical with SEQ ID NO: 97.

The DNA sequence of the PIG51 has an open reading frame encoding 247 amino acid residues, and the amino acid sequence derived from the open reading frame was

identical with SEQ ID NO: 98. The derived protein also had a molecular weight of

approximately 28 kDa.

The transformed E. coli strain was incubated in LB broth, and then 1 mM

reacted at 37 ^°C for 3 hours to express the PIG51 gene. A protein sample was

Molecular Cloning: A Laboratory manual, New York: Cold Spring Harbor Laboratory

(1989)).

FIG. 54 is a diagram showing an SDS-PAGE analysis of the PIG51 protein. In

FIG. 54, Lane 1 represents a protein sample before the IPTG induction, and Lane 2 represents a protein sample after the expression of the PIG51 gene is induced by IPTG. As shown in FIG. 54, the expressed PIG51 protein has a molecular weight of

approximately 28 kDa, which corresponds to a molecular weight of a protein derived

from its DNA sequence. A DNA base sequence result of the MIG 12 was identical with SEQ ID NO: 101.

The DNA sequence of the MIG 12 has an open reading frame encoding 44 amino acid

residues, and the amino acid sequence derived from the open reading frame was

identical with SEQ ID NO: 102. The derived protein also had a molecular weight of

approximately 5 kDa.

The transformed E. coli strain was incubated in LB broth, and then 1 mM

reacted at 37 ^°C for 3 hours to express the MIG12 gene. A protein sample was

Molecular Cloning: A Laboratory manual, New York: Cold Spring Harbor Laboratory

(1989)).

FIG. 55 is a diagram showing an SDS-PAGE analysis of the MIG 12 protein. In

FIG. 55, Lane 1 represents a protein sample before the IPTG induction, and Lane 2 represents a protein sample after the expression of the MIG 12 gene is induced by IPTG.

As shown in FIG. 55, the expressed MIGl 2 protein has a molecular weight of

approximately 5 kDa, which corresponds to a molecular weight of a protein derived

from its DNA sequence.

A DNA base sequence result of the PIG37 was identical with SEQ ID NO: 105.

The DNA sequence of the PIG37 has an open reading frame encoding 472 amino acid residues, and the amino acid sequence derived from the open reading frame was identical with SEQ ID NO: 106. The derived protein also had a molecular weight of approximately 53 kDa. The resultant full-length PIG37 cDNA was inserted into the prokaryotic expression vector pBAD/Thio-Topo (Invitrogen, U.A.), and then E. coli

which was designated E. coli DH5 α /PIG37/pBAD/Thio-Topo.

The transformed E. coli strain was incubated in LB broth, and then 1 mM

reacted at 37 ^°C for 3 hours to express the PIG37 gene. A protein sample was

sample according to the method as described in the disclosure (Sambrook, J. et al,

FIG. 56 is a diagram showing an SDS-PAGE analysis of the PIG37 protein. In

FIG. 56, Lane 1 represents a protein sample before the IPTG induction, and Lane 2

represents a protein sample after the expression of the PIG37 gene is induced by IPTG. As shown in FIG. 56, the expressed PIG37 protein has a molecular weight of approximately 53 kDa, which corresponds to a molecular weight of a protein derived

from its DNA sequence.

A DNA base sequence result of the GIG44 was identical with SEQ ID NO: 109.

The DNA sequence of the GIG44 has an open reading frame encoding 113 amino acid

residues, and the amino acid sequence derived from the open reading frame was identical with SEQ ID NO: 110. The derived protein also had a molecular weight of

approximately 12 kDa.

The transformed E. coli strain was incubated in LB broth, and then 1 mM

isopropy-1-Jβ -D-thiogalactopyranoside (IPTG) was added to the culture broth, and reacted at 37 ^°C for 3 hours to express the GIG44 gene. A protein sample was

FIG. 57 is a diagram showing an SDS-PAGE analysis of the GIG44 protein. In

FIG. 57, Lane 1 represents a protein sample before the IPTG induction, and Lane 2

represents a protein sample after the expression of the GIG44 gene is induced by IPTG.

As shown in FIG. 57, the expressed GIG44 protein has a molecular weight of approximately 12 kDa, which corresponds to a molecular weight of a protein derived from its DNA sequence.

A DNA base sequence result of the GIG31 was identical with SEQ ID NO: 113.

The DNA sequence of the GIG31 has an open reading frame encoding 211 amino acid

residues, and the amino acid sequence derived from the open reading frame was

identical with SEQ ID NO: 114. The derived protein also had a molecular weight of

approximately 24 kDa.

The transformed E. coli strain was incubated in LB broth, and then 1 mM

reacted at 37 ^°C for 3 hours to express the GIG31 gene. A protein sample was

Molecular Cloning: A Laboratory manual, New York: Cold Spring Harbor Laboratory

(1989)). FIG. 58 is a diagram showing an SDS-PAGE analysis of the GIG31 protein. In

FIG. 58, Lane 1 represents a protein sample before the IPTG induction, and Lane 2 represents a protein sample after the expression of the GIG31 gene is induced by IPTG.

As shown in FIG. 58, the expressed GIG31 protein has a molecular weight of

approximately 24 kDa, which corresponds to a molecular weight of a protein derived

from its DNA sequence.

Example 3: Northern Blotting of GIG Gene

3-1. GIG8. GIGlO, GIG13, GIG30, GIG32, GIG33. GIG34. GIG35. GIG38,

GIG39, GIG43. PIG49. PIG51. GIG44 and GIG31

In order to assess expression levels of the GIG and PIG genes, the northern

blottings were carried out, as follows.

20 βg of each of the total RNA samples obtained from the three normal breast

tissues, the three primary breast cancer tissues and the breast cancer cell line MCF-7 in

Example 1 was denatured and electrophoresized in a 1 % formaldehyde agarose gel, and then the resultant agarose gels were transferred to nylon membranes

(Boehringer-Mannheim, Germany), respectively. The nylon membranes were then

hybridized at 42 °C overnight with the ³²P-labeled random prime probes prepared from

the partial sequences FC33; FC42; FC59; FC48; FC82; FC86; FC35; FC38; FC122;

FC 126; FC 102; FClOl; FC22; FC 123 and FC47 of the full-length GIG cDNAs using the Rediprime II random prime labelling system (Amersham, United Kingdom). The northern blotting procedure was repeated twice; one is that the blots were quantitified

using the densitometer and the other is that the blots were hybridized with the β -actin probe to determine the total amount of mRNA.

FIG. 59 shows the northern blotting result that the GIG8 gene is differentially

expressed in a normal breast tissue, a primary breast cancer tissue and a breast cancer

cell line, and a bottom of FIG. 59 shows the northern blotting result obtained by

hybridizing the same blot with β -actin probe. As shown in FIG. 59 and the bottom of

FIG. 59, it was revealed that the expression level of the GIG8 gene was highly detected

all in the three samples of the normal breast tissue, but its expression was significantly

lower or not detected in the three samples of the breast cancer tissue than the normal

tissue, and very slightly detected even in the one sample of the breast cancer cell line.

The northern blotting was carried out on the normal human multiple tissue

(Clontech) and the human cancer cell line (Clontech). That is to say, the northern

blotting was carried out by hybridizing blots, to which each of the total RNA samples

extracted from the normal tissues and the cancer cell lines was transferred, in the same

manner as described above, wherein the blots were commercially available from the

company Clontech (U. S), and the normal tissue is, for example, selected from the group

consisting of brain, heart, skeletal muscle, colon, thymus, spleen, kidney, liver, small intestine, placenta, lungs and peripheral blood leukocyte, and the cancer cell line is, for example, selected from the group consisting of a promyelocytic leukemia HL-60, an

HeLa cervical cancer cell, a chronic myelocytic leukemia cell line K-562,

lymphoblastoid leukemia MOLT-4, a Burkitt's lymphoma Raji, an SW480 colon cancer cell, an A549 lung cancer cell and a G361 melanoma cell.

FIG. 88 shows a northern blotting result that the GIG8 gene is differentially expressed in various normal tissues, and a bottom of FIG. 88 shows a northern blotting result obtained by hybridizing the same blot with β -actin probe. As shown in Fig. 88,

a dominant GIG8 mRNA transcript having a size of approximately 1.3 kb was

overexpressed in the normal tissues such as the brain, the heart, the muscle, the large

the lungs and the peripheral blood. A GIG8 mRNA transcript having a size of

approximately 2.5 kb was also expressed in the normal tissues such as the liver and the peripheral blood at the same time. FIG. 117 shows a northern blotting result that the

GIG8 gene is differentially expressed in various cancer cell lines, and a bottom of FIG.

117 shows a northern blotting result obtained by hybridizing the same blot with

β -actin probe. As shown in FIG. 117, the GIG8 gene was not expressed in the tissues

such as the promyelocytic leukemia HL-60, the HeLa cervical cancer cell, the chronic

myelocytic leukemia cell line K-562, the lymphoblastoid leukemia MOLT-4, the

Burkitt's lymphoma Raji, the SW480 colon cancer cell, the A549 lung cancer cell and the G361 melanoma cell. From such a result, it might be seen that the GIG8 gene of

the present invention had the tumor suppresser function in the normal tissues such as the

breast, the brain, the heart, the muscle, the thymus, the spleen, the kidney, the liver, the small intestine, the placenta, the lungs, the large intestine and the peripheral blood.

FIG. 60 shows the northern blotting result that the GIGlO gene is differentially expressed in a normal breast tissue, a primary breast cancer tissue and a breast cancer cell line, and a bottom of FIG. 60 shows the northern blotting result obtained by

hybridizing the same blot with β -actin probe. As shown in FIG. 60, it was revealed

that the expression level of the GIGlO gene was highly detected all in the three samples

of the normal breast tissue, but its expression was significantly lower or not detected in the three samples of the breast cancer tissue than the normal tissue, and very slightly

detected even in the one sample of the breast cancer cell line.

The northern blotting was carried out on the normal human multiple tissue

company Clontech (U. S), and the normal tissue is, for example, selected from the group consisting of brain, heart, skeletal muscle, colon, thymus, spleen, kidney, liver, small

intestine, placenta, lungs and peripheral blood leukocyte, and the cancer cell line is, for

example, selected from the group consisting of a promyelocyt leukemia HL-60, an

HeLa cervical cancer cell, a chronic myelocytic leukemia cell line K-562, lymphoblastoid leukemia MOLT-4, a Burkitt's lymphoma Raji, an SW480 colon cancer

cell, an A549 lung cancer cell and a G361 melanoma cell.

FIG. 89 shows a northern blotting result that the GIGlO gene is differentially

expressed in various normal tissues, and a bottom of FIG. 89 shows a northern blotting

result obtained by hybridizing the same blot with β -actin probe. As shown in Fig. 89,

a dominant GIGlO mRNA transcript having a size of approximately 3.5 kb was

overexpressed in the normal tissues such as the brain, the heart, the muscle, the thymus, the spleen, the kidney, the liver, the small intestine, the placenta, the lungs and the peripheral blood, but not expressed in the large intestine tissue. A GIGlO mRNA transcript having a size of approximately 2.2 kb was also expressed in the normal tissues such as the heart and the placenta at the same time. FIG. 118 shows a northern blotting result that the GIGlO gene is differentially expressed in various cancer cell lines, and a

bottom of FIG. 118 shows a northern blotting result obtained by hybridizing the same

blot with β -actin probe. As shown in FIG. 118, the GIG8 gene was not expressed in

the tissues such as the HeLa cervical cancer cell, the chronic myelocytic leukemia cell

line K-562, the A549 lung cancer cell and the G361 melanoma cell, but its expression

was detected in the promyelocytic leukemia HL-60, the lymphoblastoid leukemia

MOLT-4, the Burkitt's lymphoma Raji and the SW480 colon cancer cell.

From such a result, it might be seen that the GIGlO gene of the present invention

had the tumor suppresser function in the normal tissues such as the breast, the brain, the

heart, the muscle, the thymus, the spleen, the kidney, the liver, the small intestine, the

placenta, the lungs and the peripheral blood.

FIG. 61 shows the northern blotting result that the GIG 13 gene is differentially

cell line, and a bottom of FIG. 61 shows the northern blotting result obtained by

hybridizing the same blot with β -actin probe. As shown in FIG. 61, it was revealed

that the expression level of the GIG 13 gene was highly detected all in the three samples

detected even in the one sample of the breast cancer cell line. The northern blotting was carried out on the normal human multiple tissue

extracted from the normal tissues and the cancer cell lines was transferred, in the same manner as described above, wherein the blots were commercially available from the

consisting of brain, heart, skeletal muscle, colon, thymus, spleen, kidney, liver, small

example, selected from the group consisting of a promyelocytic leukemia HL-60, an

HeLa cervical cancer cell, a chronic myelocytic leukemia cell line K-562,

lymphoblastoid leukemia MOLT-4, a Burkitt's lymphoma Raji, an SW480 colon cancer

cell, an A549 lung cancer cell and a G361 melanoma cell.

FIG. 90 shows a northern blotting result that the GIG 13 gene is differentially

expressed in various normal tissues, and a bottom of FIG. 90 shows a northern blotting

result obtained by hybridizing the same blot with β -actin probe. As shown in Fig. 90,

a dominant GIGl 3 mRNA transcript having a size of approximately 1.3 kb was overexpressed only in the normal liver tissue. FIG. 119 shows a northern blotting

result that the GIG 13 gene is differentially expressed in various cancer cell lines, and a

bottom of FIG. 119 shows a northern blotting result obtained by hybridizing the same

blot with β -actin probe. As shown in FIG. 119, the GIGl 3 gene was not expressed in

the tissues such as the promyelocytic leukemia HL-60, the HeLa cervical cancer cell, the chronic myelocytic leukemia cell line K-562, the lymphoblastoid leukemia MOLT-4, the Burkitt's lymphoma Raji, the SW480 colon cancer cell, the A549 lung cancer cell

and the G361 melanoma cell. From such a result, it might be seen that the GIG13 gene of the present invention had the tumor suppresser function in the normal tissues such as

the breast and the liver.

FIG. 67 shows the northern blotting result that the GIG30 gene is differentially expressed in a normal breast tissue, a primary breast cancer tissue and a breast cancer

cell line, and a bottom of FIG. 67 shows the northern blotting result obtained by

hybridizing the same blot with β -actin probe. As shown in FIG. 67, it was revealed

that the expression level of the GIG30 gene was highly detected all in the three samples

of the normal breast tissue, but its expression was significantly lower or not detected in

the three samples of the breast cancer tissue than the normal tissue, and very slightly

detected even in the one sample of the breast cancer cell line.

The northern blotting was carried out on the normal human multiple tissue

(Clontech) and the human cancer cell line (Clontech). That is to say, the northern blotting was carried out by hybridizing blots, to which each of the total RNA samples

consisting of brain, heart, skeletal muscle, colon, thymus, spleen, kidney, liver, small intestine, placenta, lungs and peripheral blood leukocyte, and the cancer cell line is, for example, selected from the group consisting of a promyelocytic leukemia HL-60, an HeLa cervical cancer cell, a chronic myelocytic leukemia cell line K-562,

cell, an A549 lung cancer cell and a G361 melanoma cell.

FIG. 96 shows a northern blotting result that the GIG30 gene is differentially expressed in various normal tissues, and a bottom of FIG. 96 shows a northern blotting

result obtained by hybridizing the same blot with β -actin probe. As shown in Fig. 96,

a dominant GIG30 mRNA transcript having a size of approximately 1.9 kb was overexpressed in the normal tissues such as the heart, the muscle and the liver. A

GIG30 mRNA transcript having a size of approximately 1.0 kb was also expressed in

the normal tissues such as the liver and the peripheral blood at the same time. FIG. 125 shows a northern blotting result that the GIG30 gene is differentially expressed in

various cancer cell lines, and a bottom of FIG. 125 shows a northern blotting result

obtained by hybridizing the same blot with β -actin probe. As shown in FIG. 125, the

GIG30 gene was not expressed in the tissues such as the promyelocytic leukemia HL-60,

the HeLa cervical cancer cell, the chronic myelocytic leukemia cell line K-562, the

lymphoblastoid leukemia MOLT-4, the Burkitt's lymphoma Raji, the SW480 colon

cancer cell, the A549 lung cancer cell and the G361 melanoma cell. From such a result,

it might be seen that the GIG30 gene of the present invention had the tumor suppresser

function in the normal tissues such as the breast, the heart, the muscle and the liver.

FIG. 68 shows the northern blotting result that the GIG32 gene is differentially

expressed in a normal breast tissue, a primary breast cancer tissue and a breast cancer cell line, and a bottom of FIG. 68 shows the northern blotting result obtained by

hybridizing the same blot with β -actin probe. As shown in FIG. 68, it was revealed

that the expression level of the GIG32 gene was highly detected all in the three samples of the normal breast tissue, but its expression was significantly lower or not detected in

detected even in the one sample of the breast cancer cell line.

The northern blotting was carried out on the normal human multiple tissue (Clontech) and the human cancer cell line (Clontech). That is to say, the northern blotting was carried out by hybridizing blots, to which each of the total RNA samples extracted from the normal tissues and the cancer cell lines was transferred, in the same

intestine, placenta, lungs and peripheral blood leukocyte, and the cancer cell line is, for example, selected from the group consisting of a promyelocytic leukemia HL-60, an

HeLa cervical cancer cell, a chronic myelocytic leukemia cell line K-562,

cell, an A549 lung cancer cell and a G361 melanoma cell.

FIG. 97 shows a northern blotting result that the GIG32 gene is differentially expressed in various normal tissues, and a bottom of FIG. 97 shows a northern blotting

result obtained by hybridizing the same blot with β -actin probe. As shown in Fig. 97,

a dominant GIG32 mRNA transcript having a size of approximately 4.0 kb was

overexpressed in the normal tissues such as the muscle, the large intestine, the thymus,

the spleen, the kidney, the placenta, the lungs and the peripheral blood. A GIG32

mRNA transcript having a size of approximately 1.0 kb was also expressed in the

normal tissues such as the muscle and the large intestine at the same time. FIG. 126 shows a northern blotting result that the GIG32 gene is differentially expressed in various cancer cell lines, and a bottom of FIG. 126 shows a northern blotting result

obtained by hybridizing the same blot with β -actin probe. As shown in FIG. 126, the

GIG32 gene was expressed in the tissues such as the HeLa cervical cancer cell, the A549 lung cancer cell and the G361 melanoma cell, but not expressed in the promyelocytic leukemia HL-60, the chronic myelocytic leukemia cell line K-562, the lymphoblastoid leukemia MOLT-4, the Burkitt's lymphoma Raji and the SW480 colon cancer cell.

From such a result, it might be seen that the GIG32 gene of the present invention

had the tumor suppresser function in the normal tissues such as the breast, the muscle,

the large intestine, the thymus, the spleen, the kidney, the placenta and the peripheral blood.

FIG. 70 shows the northern blotting result that the GIG34 gene is differentially

cell line, and a bottom of FIG. 70 shows the northern blotting result obtained by

hybridizing the same blot with β -actin probe. As shown in FIG. 70, it was revealed

that the expression level of the GIG34 gene was highly detected all in the three samples

detected even in the one sample of the breast cancer cell line.

The northern blotting was carried out on the normal human multiple tissue

blotting was carried out by hybridizing blots, to which each of the total RNA samples extracted from the normal tissues and the cancer cell lines was transferred, in the same manner as described above, wherein the blots were commercially available from the

intestine, placenta, lungs and peripheral blood leukocyte, and the cancer cell line is, for example, selected from the group consisting of a promyelocytic leukemia HL-60, an HeLa cervical cancer cell, a chronic myelocytic leukemia cell line K-562, lymphoblastoid leukemia MOLT-4, a Burkitt's lymphoma Raji, an SW480 colon cancer cell, an A549 lung cancer cell and a G361 melanoma cell.

FIG. 99 shows a northern blotting result that the GIG34 gene is differentially

expressed in various normal tissues, and a bottom of FIG. 99 shows a northern blotting

result obtained by hybridizing the same blot with β -actin probe. As shown in Fig. 99,

the dominant GIG34 mRNA transcript having a size of approximately 0.6 kb was

overexpressed in the normal tissues such as the brain, the heart, the muscle, the large intestine, the thymus, the spleen, the kidney, the liver, the small intestine, the placenta,

the lungs and the peripheral blood. FIG. 128 shows a northern blotting result that the

GIG34 gene is differentially expressed in various cancer cell lines, and a bottom of FIG.

128 shows a northern blotting result obtained by hybridizing the same blot with

β -actin probe. As shown in FIG. 128, the GIG34 gene was rarely expressed in the

tissues such as the promyelocytic leukemia HL-60, the HeLa cervical cancer cell, the chronic myelocytic leukemia cell line K-562, the lymphoblastoid leukemia MOLT-4, the Burkitt's lymphoma Raji, the SW480 colon cancer cell, the A549 lung cancer cell

and the G361 melanoma cell.

From such a result, it might be seen that the GIG34 gene of the present invention

heart, the muscle, the large intestine, the thymus, the spleen, the kidney, the liver, the small intestine, the placenta, the lungs and the peripheral blood.

FIG. 71 shows the northern blotting result that the GIG35 gene is differentially

expressed in a normal breast tissue, a primary breast cancer tissue and a breast cancer cell line, and a bottom of FIG. 71 shows the northern blotting result obtained by

hybridizing the same blot with β -actin probe. As shown in FIG. 71, it was revealed

that the expression level of the GIG35 gene was highly detected all in the three samples

of the normal breast tissue, but its expression was significantly lower in the three samples of the breast cancer tissue than the normal tissue, and very slightly detected

even in the one sample of the breast cancer cell line.

The northern blotting was carried out on the normal human multiple tissue

company Clontech (U.S), and the normal tissue is, for example, selected from the group

intestine, placenta, lungs and peripheral blood leukocyte, and the cancer cell line is, for example, selected from the group consisting of a promyelocyte leukemia HL-60, an

HeLa cervical cancer cell, a chronic myelocytic leukemia cell line K-562,

FIG. 100 shows a northern blotting result that the GIG35 gene is differentially

expressed in various normal tissues, and a bottom of FIG. 100 shows a northern blotting

result obtained by hybridizing the same blot with β -actin probe. As shown in Fig.

100, a GIG35 mRNA transcript having a size of approximately 1.3 kb was also expressed in the normal tissues such the brain, the heart, the muscle, the large intestine, the thymus, the spleen, the kidney, the liver, the small intestine, the placenta, the lungs

and the peripheral blood. FIG. 129 shows a northern blotting result that the GIG35

gene is differentially expressed in various cancer cell lines, and a bottom of FIG. 129

shows a northern blotting result obtained by hybridizing the same blot with β -actin

probe. As shown in FIG. 129, the GIG35 gene was rarely expressed in the tissues such

as the promyelocytic leukemia HL-60, the HeLa cervical cancer cell, the chronic myelocytic leukemia cell line K-562, the lymphoblastoid leukemia MOLT-4, the

Burkitt's lymphoma Raji, the SW480 colon cancer cell, the A549 lung cancer cell and

the G361 melanoma cell. From such a result, it might be seen that the GIG35 gene of

breast, the brain, the heart, the muscle, the thymus, the spleen, the kidney, the liver, the

small intestine, the placenta, the lungs, the large intestine and the peripheral blood.

FIG. 72 shows the northern blotting result that the GIG38 gene is differentially

cell line, and a bottom of FIG. 72 shows the northern blotting result obtained by

hybridizing the same blot with β -actin probe. As shown in FIG. 72, it was revealed

that the expression level of the GIG38 gene was highly detected all in the three samples

detected even in the one sample of the breast cancer cell line.

The northern blotting was carried out on the normal human multiple tissue

(Clontech) and the human cancer cell line (Clontech). That is to say, the northern blotting was carried out by hybridizing blots, to which each of the total RNA samples extracted from the normal tissues and the cancer cell lines was transferred, in the same

consisting of brain, heart, skeletal muscle, colon, thymus, spleen, kidney, liver, small intestine, placenta, lungs and peripheral blood leukocyte, and the cancer cell line is, for

HeLa cervical cancer cell, a chronic myelocytic leukemia cell line K-562,

cell, an A549 lung cancer cell and a G361 melanoma cell. FIG. 101 shows a northern blotting result that the GIG38 gene is differentially expressed in various normal tissues, and a bottom of FIG. 101 shows a northern blotting

101, a dominant GIG38 mRNA transcript having a size of approximately 0.7 kb was

overexpressed in the normal tissues such as the heart, the muscle, the kidney, the liver

and the placenta. GIG38 mRNA transcripts having a size of approximately 1.5 kb and

2.0 kb were also expressed in the normal tissues such as the heart and the muscle at the same time. FIG. 130 shows a northern blotting result that the GIG38 gene is

differentially expressed in various cancer cell lines, and a bottom of FIG. 130 shows a

northern blotting result obtained by hybridizing the same blot with β -actin probe. As

shown in FIG. 130, the GIG38 gene was very rarely expressed or hardly expressed in the tissues such as the promyelocytic leukemia HL-60, the HeLa cervical cancer cell, the chronic myelocytic leukemia cell line K-562, the lymphoblastoid leukemia MOLT-4,

the Burkitt's lymphoma Raji, the SW480 colon cancer cell, the A549 lung cancer cell and the G361 melanoma cell. The GIG38 mRNA transcripts having a size of

approximately 1.5 kb and 2.0 kb proven to be expressed in the normal tissues all were

not expressed in the cancer cell lines. From such a result, it might be seen that the

GIG38 gene of the present invention had the tumor suppresser function in the normal

tissues such as the breast, the heart, the muscle, the kidney, the liver and the placenta.

FIG. 73 shows the northern blotting result that the GIG39 gene is differentially

expressed in a normal breast tissue, a primary breast cancer tissue and a breast cancer cell line, and a bottom of FIG. 73 shows the northern blotting result obtained by

hybridizing the same blot with β -actin probe. As shown in FIG. 73, it was revealed

that the expression level of the GIG39 gene was highly detected all in the three samples

the three samples of the breast cancer tissue than the normal tissue, and very slightly detected even in the one sample of the breast cancer cell line.

The northern blotting was carried out on the normal human multiple tissue

blotting was carried out by hybridizing blots, to which each of the total RNA samples extracted from the normal tissues and the cancer cell lines was transferred, in the same manner as described above, wherein the blots were commercially available from the company Clontech (U. S), and the normal tissue is, for example, selected from the group consisting of brain, heart, skeletal muscle, colon, thymus, spleen, kidney, liver, small intestine, placenta, lungs and peripheral blood leukocyte, and the cancer cell line is, for

example, selected from the group consisting of a promyelocyte leukemia HL-60, an

HeLa cervical cancer cell, a chronic myelocytic leukemia cell line K-562, lymphoblastoid leukemia MOLT-4, a Burkitt's lymphoma Raji, an SW480 colon cancer cell, an A549 lung cancer cell and a G361 melanoma cell.

FIG. 102 shows a northern blotting result that the GIG39 gene is differentially

expressed in various normal tissues, and a bottom of FIG. 102 shows a northern blotting

102, a dominant GIG39 mRNA transcript having a size of 2.4 kb was overexpressed

only in the liver normal tissue. FIG. 131 shows a northern blotting result that the GIG39 gene is differentially expressed in various cancer cell lines, and a bottom of FIG.

131 shows a northern blotting result obtained by hybridizing the same blot with

β -actin probe. As shown in FIG. 131, the GIG39 gene was not expressed in the

tissues such as the promyelocytic leukemia HL-60, the HeLa cervical cancer cell, the

chronic myelocytic leukemia cell line K-562, the lymphoblastoid leukemia MOLT-4,

the Burkitt's lymphoma Raji, the SW480 colon cancer cell, the A549 lung cancer cell

and the G361 melanoma cell. From such a result, it might be seen that the GIG39 gene

of the present invention had the tumor suppresser function in the normal tissues such as the breast and the liver.

FIG. 76 shows the northern blotting result that the GIG43 gene is differentially expressed in a normal breast tissue, a primary breast cancer tissue and a breast cancer

cell line, and a bottom of FIG. 76 shows the northern blotting result obtained by

hybridizing the same blot with β -actin probe. As shown in FIG. 76, it was revealed

that the expression level of the GIG43 gene was highly detected all in the three samples

The northern blotting was carried out on the normal human multiple tissue

manner as described above, wherein the blots were commercially available from the company Clontech (U. S), and the normal tissue is, for example, selected from the group

HeLa cervical cancer cell, a chronic myelocytic leukemia cell line K-562,

cell, an A549 lung cancer cell and a G361 melanoma cell.

FIG. 105 shows a northern blotting result that the GIG43 gene is differentially

expressed in various normal tissues, and a bottom of FIG. 105 shows a northern blotting

105, the dominant GIG43 mRNA transcript having a size of approximately 3.5 kb was

overexpressed in the normal tissues such as the heart, the kidney, the liver, the placenta and the lungs. FIG. 134 shows a northern blotting result that the GIG43 gene is

differentially expressed in various cancer cell lines, and a bottom of FIG. 134 shows a

shown in FIG. 134, the GIG8 gene was not expressed in the tissues such as the promyelocytic leukemia HL-60, the HeLa cervical cancer cell, the chronic myelocytic leukemia cell line K-562, the lymphoblastoid leukemia MOLT-4, the Burkitt's

lymphoma Raji, the SW480 colon cancer cell, the A549 lung cancer cell and the G361

melanoma cell. From such a result, it might be seen that the GIG8 gene of the present

invention had the tumor suppresser function in the normal tissues such as the breast, the

heart, the kidney, the liver, the placenta and the lungs.

FIG. 82 shows the northern blotting result that the PIG49 gene is differentially

expressed in a normal breast tissue, a primary breast cancer tissue and a breast cancer cell line, and a bottom of FIG. 82 shows the northern blotting result obtained by

hybridizing the same blot with β -actin probe. As shown in FIG. 82, it was revealed

that the expression level of the PIG49 gene was highly detected all in the three samples

detected even in the one sample of the breast cancer cell line.

The northern blotting was carried out on the normal human multiple tissue (Clontech) and the human cancer cell line (Clontech). That is to say, the northern

company Clontech (U.S), and the normal tissue is, for example, selected from the group consisting of brain, heart, skeletal muscle, colon, thymus, spleen, kidney, liver, small intestine, placenta, lungs and peripheral blood leukocyte, and the cancer cell line is, for example, selected from the group consisting of a promyelocytic leukemia HL-60, an

cell, an A549 lung cancer cell and a G361 melanoma cell.

FIG. I ll shows a northern blotting result that the PIG49 gene is differentially

expressed in various normal tissues, and a bottom of FIG. I l l shows a northern blotting

I l l, a dominant PIG49 mRNA transcript having a size of approximately 2.4 kb was

and the placenta. A PIG49 mRNA transcript having a size of approximately 1.5 kb was also expressed in the normal muscle tissue at the same time. FIG. 140 shows a

northern blotting result that the PIG49 gene is differentially expressed in various cancer

cell lines, and a bottom of FIG. 140 shows a northern blotting result obtained by

hybridizing the same blot with β -actin probe. As shown in FIG. 140, the PIG49 gene

was not expressed or very rarely expressed in the tissues such as the promyelocytic leukemia HL-60, the lymphoblastoid leukemia MOLT-4, the HeLa cervical cancer cell,

the chronic myelocytic leukemia cell line K-562, the lymphoblastoid leukemia MOLT-4, the Burkitt's lymphoma Raji, the SW480 colon cancer cell, the A549 lung cancer cell and the G361 melanoma cell.

From such a result, it might be seen that the PIG49 gene of the present invention

had the tumor suppresser function in the normal tissues such as the breast, the heart, the

muscle, the kidney, the liver and the placenta.

FIG. 83 shows the northern blotting result that the PIG51 gene is differentially

expressed in a normal breast tissue, a primary breast cancer tissue and a breast cancer cell line, and a bottom of FIG. 83 shows the northern blotting result obtained by hybridizing the same blot with β -actin probe. As shown in FIG. 83, it was revealed

that the expression level of the PIG51 gene was highly detected all in the three samples

detected even in the one sample of the breast cancer cell line.

HeLa cervical cancer cell, a chronic myelocytic leukemia cell line K-562,

FIG. 112 shows a northern blotting result that the PIG51 gene is differentially

expressed in various normal tissues, and a bottom of FIG. 112 shows a northern blotting

112, a dominant PIG51 mRNA transcript having a size of approximately 1.0 kb was overexpressed in the normal tissues such as the heart, the muscls, the thymus, the spleen,

the kidney, the liver, the placenta and the peripheral blood. FIG. 141 shows a northern blotting result that the PIG51 gene is differentially expressed in various cancer cell lines,

and a bottom of FIG. 141 shows a northern blotting result obtained by hybridizing the

same blot with β -actin probe. As shown in FIG. 141, the PIG51 gene was not

expressed in the tissues such as the promyelocyte leukemia HL-60, the HeLa cervical

cancer cell, the chronic myelocytic leukemia cell line K-562, the lymphoblastoid

leukemia MOLT-4, the Burkitt's lymphoma Raji, the SW480 colon cancer cell, the A549 lung cancer cell and the G361 melanoma cell. From such a result, it might be seen that the PIG51 gene of the present invention had the tumor suppresser function in

the normal tissues such as the breast, the heart, the muscle, the thymus, the spleen, the

kidney, the liver, the placenta and the peripheral blood.

FIG. 86 shows the northern blotting result that the GIG44 gene is differentially

cell line, and a bottom of FIG. 86 shows the northern blotting result obtained by

hybridizing the same blot with β -actin probe. As shown in FIG. 86, it was revealed

that the expression level of the GIG44 gene was highly detected all in the three samples

HeLa cervical cancer cell, a chronic myelocytic leukemia cell line K-562,

cell, an A549 lung cancer cell and a G361 melanoma cell.

FIG. 115 shows a northern blotting result that the GIG44 gene is differentially expressed in various normal tissues, and a bottom of FIG. 115 shows a northern blotting

115, a dominant GIG44 mRNA transcript having a size of approximately 1.0 kb was

the lung and the leukocyte. A GIG44 mRNA transcript having a size of approximately

0.5 kb was also expressed in the normal tissues at the same time.

FIG. 144 shows a northern blotting result that the GIG44 gene is differentially

expressed in various cancer cell lines, and a bottom of FIG. 144 shows a northern

blotting result obtained by hybridizing the same blot with β -actin probe. As shown

in FIG. 144, the GIG44 gene was very rarely expressed in the tissues such as the promyelocytic leukemia HL-60, the HeLa cervical cancer cell, the chronic myelocytic leukemia cell line K-562, the lymphoblastoid leukemia MOLT-4, the Burkitt's

lymphoma Raji, the SW480 colon cancer cell, the A549 lung cancer cell and the G361 melanoma cell. From such a result, it might be seen that the GIG44 gene of the present invention had the tumor suppresser function in the normal tissues such as the brain, the

heart, the muscle, the large intestine, the thymus, the spleen, the kidney, the liver, the

small intestine, the placenta, the lung and the leukocyte.

FIG. 87 shows the northern blotting result that the GIG31 gene is differentially

cell line, and a bottom of FIG. 87 shows the northern blotting result obtained by

hybridizing the same blot with β -actin probe. As shown in FIG. 87, it was revealed

that the expression level of the GIG31 gene was highly detected all in the three samples

detected even in the one sample of the breast cancer cell line.

manner as described above, wherein the blots were commercially available from the company Clontech (U.S), and the normal tissue is, for example, selected from the group consisting of brain, heart, skeletal muscle, colon, thymus, spleen, kidney, liver, small

example, selected from the group consisting of a promyelocytic leukemia HL-60, an HeLa cervical cancer cell, a chronic myelocytic leukemia cell line K-562, lymphoblastoid leukemia MOLT-4, a Burkitt's lymphoma Raji, an SW480 colon cancer

cell, an A549 lung cancer cell and a G361 melanoma cell. FIG. 116 shows a northern blotting result that the GIG31 gene is differentially

expressed in various normal tissues, and a bottom of FIG. 116 shows a northern blotting

116, a dominant GIG31 mRNA transcript having a size of approximately 1.4 kb was

overexpressed in the normal tissues such as the breast, the heart, the large intestine, the

spleen, the small intestine, the placenta, the lung and the leukocyte. FIG. 145 shows a northern blotting result that the GIG31 gene is differentially expressed in various cancer

cell lines, and a bottom of FIG. 145 shows a northern blotting result obtained by

hybridizing the same blot with β -actin probe. As shown in FIG. 145, the GIG31 gene

was not expressed in the tissues such as the promyelocytic leukemia HL-60, the chronic

myelocytic leukemia cell line K-562, the lymphoblastoid leukemia MOLT-4, the

Burkitt's lymphoma Raji, the SW480 colon cancer cell and the G361 melanoma cell, but

very rarely expressed in the HeLa cervical cancer cell and the A549 lung cancer cell. From such a result, it might be seen that the GIG31 gene of the present invention had the tumor suppresser function in the normal tissues such as the breast, the heart, the

large intestine, the spleen, the small intestine, the placenta, the lung and the leukocyte.

3-2. GIG15

In order to assess an expression level of the GIG 15 gene, the northern blotting

was carried out, as follows. The total RNA samples were extracted from the normal bone marrow tissue, the leukemia bone marrow tissue and the K-562 cell, as described

in Example 1. 20 μg of each of the total RNA samples was denatured and

electrophoresized in a 1 % formaldehyde agarose gel, and then the resultant agarose gels were transferred to nylon membranes (Boehringer-Mannheim, Germany), respectively. The nylon membranes were then hybridized at 42 ⁰C overnight with the ³²P-labeled

random prime probes prepared from the partial sequence GV2 of the full-length GIGl 5

cDNA using the Rediprime II random prime labelling system (Amersham, United Kingdom). The northern blotting procedure was repeated twice; one is that the blots

were quantitified using the densitometer and the other is that the blots were hybridized

with the β -actin probe to determine the total amount of mRNA.

FIG. 62 shows the northern blotting result that the GIG 15 gene is differentially

expressed in a normal bone marrow tissue, a leukemia bone marrow tissue and a K-562

cell, and a bottom of FIG. 62 shows the northern blotting result obtained by hybridizing

the same blot with β -actin probe. As shown in FIG. 62, it was revealed that the

expression level of the GIGl 5 gene was highly detected all in the samples of the normal

bone marrow tissue, but its expression was significantly lower in the samples of the

leukemia bone marrow tissue than the normal tissue, and slightly detected even in the

one sample of the leukemia cell line. The northern blotting was carried out on the normal human multiple tissue

manner as described above, wherein the blots were commercially available from the company Clontech (U. S), and the normal tissue is, for example, selected from the group consisting of brain, heart, skeletal muscle, colon, thymus, spleen, kidney, liver, small

intestine, placenta, lungs and peripheral blood leukocyte, and the cancer cell line is, for example, selected from the group consisting of a promyelocytic leukemia HL-60, an HeLa cervical cancer cell, a chronic myelocytic leukemia cell line K-562,

cell, an A549 lung cancer cell and a G361 melanoma cell.

FIG. 91 shows a northern blotting result that the GIGl 5 gene is differentially

expressed in various normal tissues, and a bottom of FIG. 91 shows a northern blotting

result obtained by hybridizing the same blot with β -actin probe. As shown in Fig. 91,

a dominant GIG 15 mRNA transcript having a size of approximately 0.5 kb was

the lung and the peripheral blood. A GIG 15 mRNA transcript having a size of

approximately 1.0 kb was also expressed in the normal tissues such as the liver and the

kidney at the same time. FIG. 120 shows a northern blotting result that the GIG15

gene is differentially expressed in various cancer cell lines, and a bottom of FIG. 120

probe. As shown in FIG. 120, the GIGl 5 gene was rarely expressed in the tissues such

as the promyelocytic leukemia HL-60, the HeLa cervical cancer cell, the chronic

myelocytic leukemia cell line K-562, the lymphoblastoid leukemia MOLT-4, the

the G361 melanoma cell. From such a result, it might be seen that the GIG 15 gene of the present invention had the tumor suppresser function in the normal tissues such as the bone marrow, the brain, the heart, the muscle, the thymus, the spleen, the kidney, the

liver, the small intestine, the placenta, the lung, the large intestine and the peripheral

blood. 3-3. GIGl6. GIG24. GIG26, GIG29, GIG40. GIG42. PIG33. PIG35. PIG36. PIG37

In order to assess an expression level of the GIG and PIG genes, the northern blottings were carried out, as follows.

20 βg of each of the total RNA samples obtained from the three normal liver

tissues, the three primary liver cancer tissues and the liver cancer cell line HepG2 in

Example 1 was denatured and electrophoresized in a 1 % formaldehyde agarose gel, and

then the resultant agarose gels were transferred to nylon membranes

(Boehringer-Mannheim, Germany), respectively. The nylon membranes were then

hybridized at 42 ^°C overnight with the ³²P-labeled random prime probes prepared from

the full-length GIG and PIG cDNAs using the Rediprime II random prime labelling

system (Amersham, United Kingdom). The northern blotting procedure was repeated

twice; one is that the blots were quantitified using the densitometer and the other is that

the blots were hybridized with the β -actin probe to determine the total amount of

mRNA.

FIG. 63 shows the northern blotting result that the GIG 16 gene is differentially expressed in a normal liver tissue, a primary liver cancer tissue and a liver cancer cell line, and a bottom of FIG. 63 shows the northern blotting result obtained by hybridizing

the same blot with β -actin probe. As shown in FIG. 63, it was revealed that the

expression level of the GIG 16 gene was highly detected all in the three samples of the normal liver tissue, but its expression was significantly lower in the three samples of the

liver cancer tissue than the normal tissue, and not detected in the one sample of the liver

cancer cell line. The northern blotting was carried out on the normal human multiple tissue

cell, an A549 lung cancer cell and a G361 melanoma cell.

FIG. 92 shows a northern blotting result that the GIGl 6 gene is differentially

expressed in various normal tissues, and a bottom of FIG. 92 shows a northern blotting

result obtained by hybridizing the same blot with β -actin probe. As shown in Fig. 92,

the dominant GIG 16 mRNA transcript having a size of approximately 2.0 kb was

overexpressed in the normal tissues such as the liver and the kidney.

FIG. 121 shows a northern blotting result that the GIG 16 gene is differentially expressed in various cancer cell lines, and a bottom of FIG. 121 shows a northern

in FIG. 121, the GIG 16 gene was not expressed at all in the tissues such as the

promyelocytic leukemia HL-60, the HeLa cervical cancer cell, the chronic myelocytic

leukemia cell line K-562, the lymphoblastoid leukemia MOLT-4, the Burkitt's lymphoma Raji, the SW480 colon cancer cell, the A549 lung cancer cell and the G361

melanoma cell. From such a result, it might be seen that the GIGl 6 gene of the present

invention had the tumor suppresser function in the normal tissues such as the liver and

the kidney. Also, it might be seen that the GIG 16 gene of the present invention had the

tumor suppresser function from the fact that its expression was suppressed even in the

leukemia, the uterine cancer, the malignant lymphoma, the colon cancer, the lung cancer

and the skin cancer to induce tumorigenesis.

FIG. 64 shows the northern blotting result that the GIG24 gene is differentially

line, and a bottom of FIG. 64 shows the northern blotting result obtained by hybridizing

the same blot with β -actin probe. As shown in FIG. 64, it was revealed that the

expression level of the GIG24 gene was highly detected all in the three samples of the

normal liver tissue, but its expression was significantly lower in the three samples of the liver cancer tissue than the normal tissue, and not detected in the one sample of the liver

cancer cell line.

The northern blotting was carried out on the normal human multiple tissue

manner as described above, wherein the blots were commercially available from the company Clontech (U. S), and the normal tissue is, for example, selected from the group consisting of brain, heart, skeletal muscle, colon, thymus, spleen, kidney, liver, small intestine, placenta, lungs and peripheral blood leukocyte, and the cancer cell line is, for example, selected from the group consisting of a promyelocyte leukemia HL-60, an

HeLa cervical cancer cell, a chronic myelocytic leukemia cell line K-562,

cell, an A549 lung cancer cell and a G361 melanoma cell.

FIG. 93 shows a northern blotting result that the GIG24 gene is differentially

expressed in various normal tissues, and a bottom of FIG. 93 shows a northern blotting

result obtained by hybridizing the same blot with β -actin probe. As shown in Fig. 93,

the dominant GIG24 mRNA transcript having a size of approximately 2.4 kb was

overexpressed in the normal tissues such as the liver, the heart and the muscle. FIG. 122 shows a northern blotting result that the GIG24 gene is differentially

expressed in various cancer cell lines, and a bottom of FIG. 122 shows a northern

in FIG. 122, the GIG24 gene was not expressed at all in the tissues such as the

promyelocytic leukemia HL-60, the HeLa cervical cancer cell, the chronic myelocytic leukemia cell line K-562, the lymphoblastoid leukemia MOLT-4, the Burkitt's

melanoma cell. From such a result, it might be seen that the GIG24 gene of the present invention had the tumor suppresser function in the normal tissues such as the liver, the

heart and the muscle. Also, it might be seen that the GIG24 gene of the present

invention had the tumor suppresser function from the fact that its expression was suppressed even in the leukemia, the uterine cancer, the malignant lymphoma, the colon

cancer, the lung cancer and the skin cancer to induce tumorigenesis.

FIG. 65 shows the northern blotting result that the GIG26 gene is differentially expressed in a normal liver tissue, a primary liver cancer tissue and a liver cancer cell

line, and a bottom of FIG. 65 shows the northern blotting result obtained by hybridizing

the same blot with β -actin probe. As shown in FIG. 65, it was revealed that the

expression level of the GIG26 gene was highly detected all in the three samples of the

normal liver tissue, but its expression was significantly lower in the three samples of the

liver cancer tissue than the normal tissue, and not detected in the one sample of the liver cancer cell line.

The northern blotting was carried out on the normal human multiple tissue

extracted from the normal tissues and the cancer cell lines was transferred, in the same manner as described above, wherein the blots were commercially available from the company Clontech (U. S), and the normal tissue is, for example, selected from the group

HeLa cervical cancer cell, a chronic myelocytic leukemia cell line K-562, lymphoblastoid leukemia MOLT-4, a Burkitt's lymphoma Raji, an SW480 colon cancer cell, an A549 lung cancer cell and a G361 melanoma cell. FIG. 94 shows a northern blotting result that the GIG26 gene is differentially expressed in various normal tissues, and a bottom of FIG. 94 shows a northern blotting

result obtained by hybridizing the same blot with β -actin probe. As shown in Fig. 94,

the dominant GIG26 mRNA transcript having a size of approximately 2.0 kb was overexpressed in the normal liver tissue, and GIG26 mRNA transcripts having a size of

approximately 2.5 kb and 1.5 kb were also expressed in the kidney, the brain and the

heart at the same time. FIG. 123 shows a northern blotting result that the GIG26 gene

is differentially expressed in various cancer cell lines, and a bottom of FIG. 123 shows a

shown in FIG. 123, the GIG26 gene was not expressed at all in the tissues such as the

leukemia cell line K-562, the lymphoblastoid leukemia MOLT-4, the Burkitt's

melanoma cell. From such a result, it might be seen that the GIG26 gene of the present

invention had the tumor suppresser function in the normal tissues such as the liver, the

kidney, the brain and the heart. Also, it might be seen that the GIG 16 gene of the

present invention had the tumor suppresser function from the fact that its expression was

suppressed even in the leukemia, the uterine cancer, the malignant lymphoma, the colon

cancer, the lung cancer and the skin cancer to induce tumorigenesis.

FIG. 66 shows the northern blotting result that the GIG29 gene is differentially expressed in a normal liver tissue, a primary liver cancer tissue and a liver cancer cell

line, and a bottom of FIG. 66 shows the northern blotting result obtained by hybridizing

the same blot with β -actin probe. As shown in FIG. 66, it was revealed that the

expression level of the GIG29 gene was highly detected all in the three samples of the normal liver tissue, but its expression was significantly lower in the three samples of the liver cancer tissue than the normal tissue, and not detected in the one sample of the liver

example, selected from the group consisting of a promyelocytic leukemia HL-60, an HeLa cervical cancer cell, a chronic myelocytic leukemia cell line K-562,

cell, an A549 lung cancer cell and a G361 melanoma cell.

FIG. 95 shows a northern blotting result that the GIG29 gene is differentially

expressed in various normal tissues, and a bottom of FIG. 95 shows a northern blotting

the dominant GIG29 mRNA transcript having a size of approximately 1.4 kb was overexpressed in the normal liver tissue.

FIG. 124 shows a northern blotting result that the GIG29 gene is differentially

expressed in various cancer cell lines, and a bottom of FIG. 124 shows a northern

in FIG. 124, the GIG29 gene was not expressed at all in the tissues such as the promyelocytic leukemia HL-60, the HeLa cervical cancer cell, the chronic myelocytic leukemia cell line K-562, the lymphoblastoid leukemia MOLT-4, the Burkitt's lymphoma Raji, the SW480 colon cancer cell, the A549 lung cancer cell and the G361

melanoma cell. From such a result, it might be seen that the GIG29 gene of the present

invention had the tumor suppresser function in the normal liver tissue. Also, it might

be seen that the GIG29 gene of the present invention had the tumor suppresser function

from the fact that its expression was suppressed even in the leukemia, the uterine cancer,

the malignant lymphoma, the colon cancer, the lung cancer and the skin cancer to induce tumorigenesis.

FIG. 74 shows the northern blotting result that the GIG40 gene is differentially

line, and a bottom of FIG. 74 shows the northern blotting result obtained by hybridizing

the same blot with β -actin probe. As shown in FIG. 74, it was revealed that the

expression level of the GIG40 gene was highly detected all in the three samples of the

cancer cell line.

The northern blotting was carried out on the normal human multiple tissue

blotting was carried out by hybridizing blots, to which each of the total RNA samples extracted from the normal tissues and the cancer cell lines was transferred, in the same manner as described above, wherein the blots were commercially available from the company Clontech (U. S), and the normal tissue is, for example, selected from the group

HeLa cervical cancer cell, a chronic myelocytic leukemia cell line K-562,

FIG. 103 shows a northern blotting result that the GIG40 gene is differentially

expressed in various normal tissues, and a bottom of FIG. 103 shows a northern blotting

103, the dominant GIG40 mRNA transcript having a size of approximately 1.5 kb was

overexpressed in the normal tissues such as the liver, the heart and the muscle. A

GIG40 mRNA transcript having a size of approximately 5.0 kb was expressed at the

same time.

FIG. 132 shows a northern blotting result that the GIG40 gene is differentially

expressed in various cancer cell lines, and a bottom of FIG. 132 shows a northern

in FIG. 132, the GIG40 gene was very rarely expressed in the tissues such as the promyelocytic leukemia HL-60, the HeLa cervical cancer cell, the chronic myelocytic

leukemia cell line K-562, the lymphoblastoid leukemia MOLT-4, the Burkitt's

melanoma cell. From such a result, it might be seen that the GIG40 gene of the present invention had the tumor suppresser function in the normal tissues such as the liver, the heart and the muscle. Also, it might be seen that the GIG40 gene of the present invention had the tumor suppresser function from the fact that its expression was suppressed even in the leukemia, the uterine cancer, the malignant lymphoma, the colon cancer, the lung cancer and the skin cancer to induce tumorigenesis.

FIG. 75 shows the northern blotting result that the GIG42 gene is differentially

line, and a bottom of FIG. 75 shows the northern blotting result obtained by hybridizing

the same blot with β -actin probe. As shown in FIG. 75, it was revealed that the

expression level of the GIG42 gene was highly detected all in the three samples of the

cancer cell line.

The northern blotting was carried out on the normal human multiple tissue

FIG. 104 shows a northern blotting result that the GIG42 gene is differentially

expressed in various normal tissues, and a bottom of FIG. 104 shows a northern blotting result obtained by hybridizing the same blot with β -actin probe. As shown in Fig.

104, the dominant GIG42 mRNA transcript having a size of approximately 2.5 kb was overexpressed in the normal liver tissue.

FIG. 133 shows a northern blotting result that the GIG42 gene is differentially

expressed in various cancer cell lines, and a bottom of FIG. 133 shows a northern

in FIG. 133, the GIG42 gene was not expressed at all in the tissues such as the promyelocyte leukemia HL-60, the HeLa cervical cancer cell, the chronic myelocytic

leukemia cell line K-562, the lymphoblastoid leukemia MOLT-4, the Burkitt's

melanoma cell. From such a result, it might be seen that the GIG42 gene of the present

be seen that the GIG42 gene of the present invention had the tumor suppresser function

from the fact that its expression was suppressed even in the leukemia, the uterine cancer, the malignant lymphoma, the colon cancer, the lung cancer and the skin cancer to induce

tumorigenesis.

FIG. 78 shows the northern blotting result that the PIG33 gene is differentially

expressed in a normal liver tissue, a primary liver cancer tissue and a liver cancer cell line, and a bottom of FIG. 78 shows the northern blotting result obtained by hybridizing

the same blot with β -actin probe. As shown in FIG. 78, it was revealed that the

expression level of the PIG33 gene was highly detected all in the three samples of the

normal liver tissue, but its expression was significantly lower in the three samples of the liver cancer tissue than the normal tissue, and not detected in the one sample of the liver cancer cell line.

The northern blotting was carried out on the normal human multiple tissue

cell, an A549 lung cancer cell and a G361 melanoma cell.

FIG. 107 shows a northern blotting result that the PIG33 gene is differentially expressed in various normal tissues, and a bottom of FIG. 107 shows a northern blotting

107, the dominant PIG33 mRNA transcript having a size of approximately 3.0 kb was

overexpressed in the normal tissues such as the brain, the heart, the skeletal muscle, the

colon, the thymus, the spleen, the kidney, the liver, the small intestine, the placenta and

the lung.

FIG. 136 shows a northern blotting result that the PIG33 gene is differentially expressed in various cancer cell lines, and a bottom of FIG. 136 shows a northern

blotting result obtained by hybridizing the same blot with β -actin probe. As shown in FIG. 136, the PIG33 gene was not expressed at all in the tissues such as the HeLa

cervical cancer cell, the chronic myelocytic leukemia cell line K-562, the

lymphoblastoid leukemia MOLT-4, the SW480 colon cancer cell, the A549 lung cancer

cell and the G361 melanoma cell and rarely expressed in the promyelocytic leukemia

HL-60 and the Burkitt's lymphoma Raji. From such a result, it might be seen that the PIG33 gene of the present invention had the tumor suppresser function in the normal

tissues such as the brain, the heart, the skeletal muscle, the colon, the thymus, the spleen, the kidney, the liver, the small intestine, the placenta and the lung. Also, it might be

seen that the PIG33 gene of the present invention had the tumor suppresser function

the colon cancer, the lung cancer and the skin cancer to induce tumorigenesis.

FIG. 79 shows the northern blotting result that the PIG35 gene is differentially

expressed in a normal liver tissue, a primary liver cancer tissue and a liver cancer cell line, and a bottom of FIG. 79 shows the northern blotting result obtained by hybridizing

the same blot with β -actin probe. As shown in FIG. 79, it was revealed that the

expression level of the PIG35 gene was highly detected all in the three samples of the normal liver tissue, but its expression was significantly lower in the three samples of the

HeLa cervical cancer cell, a chronic myelocytic leukemia cell line K-562,

cell, an A549 lung cancer cell and a G361 melanoma cell.

FIG. 108 shows a northern blotting result that the PIG35 gene is differentially

expressed in various normal tissues, and a bottom of FIG. 108 shows a northern blotting

108, the dominant PIG35 mRNA transcript having a size of approximately 1.7 kb was

liver, the small intestine, the placenta and the lungs. FIG. 137 shows a northern blotting result that the PIG35 gene is differentially expressed in various cancer cell lines, and a bottom of FIG. 137 shows a northern

in FIG. 137, the PIG35 gene was not expressed at all in the tissues such as the

promyelocytic leukemia HL-60, the HeLa cervical cancer cell, the lymphoblastoid leukemia MOLT-4, the Burkitt's lymphoma Raji, the SW480 colon cancer cell, the A549 lung cancer cell and the G361 melanoma cell, but rarely expressed in the chronic myelocytic leukemia cell line K-562. From such a result, it might be seen that the PIG35 gene of the present invention had the tumor suppresser function in the normal tissues such as the brain, the heart, the skeletal muscle, the liver, the small intestine, the

placenta and the lungs. Also, it might be seen that the PIG35 gene of the present

cancer, the lung cancer and the skin cancer to induce tumorigenesis.

FIG. 80 shows the northern blotting result that the PIG36 gene is differentially

line, and a bottom of FIG. 80 shows the northern blotting result obtained by hybridizing

the same blot with β -actin probe. As shown in FIG. 80, it was revealed that the

expression level of the PIG36 gene was highly detected all in the three samples of the

cancer cell line.

The northern blotting was carried out on the normal human multiple tissue

example, selected from the group consisting of a promyelocyt leukemia HL-60, an HeLa cervical cancer cell, a chronic myelocytic leukemia cell line K-562, lymphoblastoid leukemia MOLT-4, a Burkitt's lymphoma Raji, an SW480 colon cancer

cell, an A549 lung cancer cell and a G361 melanoma cell.

FIG. 109 shows a northern blotting result that the PIG36 gene is differentially

expressed in various normal tissues, and a bottom of FIG. 109 shows a northern blotting

109, the dominant PIG36 mRNA transcript having a size of approximately 1.0 kb was overexpressed in the normal liver tissue.

FIG. 138 shows a northern blotting result that the PIG36 gene is differentially

expressed in various cancer cell lines, and a bottom of FIG. 138 shows a northern

in FIG. 138, the PIG36 gene was not expressed at all or rarely expressed in the tissues

such as the promyelocyte leukemia HL-60, the HeLa cervical cancer cell, the chronic

myelocytic leukemia cell line K-562, the lymphoblastoid leukemia MOLT-4, the Burkitt's lymphoma Raji, the SW480 colon cancer cell, the A549 lung cancer cell and

the G361 melanoma cell. From such a result, it might be seen that the PIG36 gene of the present invention had the tumor suppresser function in the normal tissues such as the

liver, the heart, the muscle, the kidney and the placenta. Also, it might be seen that the PIG36 gene of the present invention had the tumor suppresser function from the fact that

its expression was suppressed even in the leukemia, the uterine cancer, the malignant lymphoma, the colon cancer, the lung cancer and the skin cancer to induce tumorigenesis.

FIG. 85 shows the northern blotting result that the PIG37 gene is differentially

expressed in a normal liver tissue, a primary liver cancer tissue and a liver cancer cell line, and a bottom of FIG. 85 shows the northern blotting result obtained by hybridizing

the same blot with β -actin probe. As shown in FIG. 85, it was revealed that the

expression level of the PIG37 gene was highly detected all in the three samples of the

cancer cell line.

The northern blotting was carried out on the normal human multiple tissue

blotting was carried out by hybridizing blots, to which each of the total RNA samples extracted from the normal tissues and the cancer cell lines was transferred, in the same

FIG. 114 shows a northern blotting result that the PIG37 gene is differentially

expressed in various normal tissues, and a bottom of FIG. 114 shows a northern blotting

114, the dominant PIG37 mRNA transcript having a size of approximately 7.0 kb was overexpressed in the normal tissues such as the brain, the heart, the skeletal muscle, the colon, the thymus, the spleen, the kidney, the liver, the small intestine, the placenta and

the lung. PIG37 mRNA transcripts having a size of approximately 2.0 and 1.0 kb were

overexpressed in the normal tissues at the same time.

FIG. 143 shows a northern blotting result that the PIG37 gene is differentially

expressed in various cancer cell lines, and a bottom of FIG. 143 shows a northern

in FIG. 143, the PIG37 gene was hardly expressed in the tissues such as the

promyelocytic leukemia HL-60, the chronic myelocytic leukemia cell line K-562, the

lymphoblastoid leukemia MOLT-4, the SW480 colon cancer cell and the A549 lung

cancer cell, but rarely expressed in the HeLa cervical cancer cell, the Burkitt's lymphoma Raji and the G361 melanoma cell. From such a result, it might be seen that

the PIG37 gene of the present invention had the tumor suppresser function in the normal

tissues such as the brain, the heart, the skeletal muscle, the colon, the thymus, the spleen,

the kidney, the liver, the small intestine, the placenta and the lungs. Also, it might be seen that the PIG37 gene of the present invention had the tumor suppresser function from the fact that its expression was suppressed even in the leukemia, the uterine cancer,

the colon cancer, the lung cancer and the skin cancer to induce tumorigenesis.

3-4. GIG46. MIG20

In order to assess an expression level of the GIG and PIG genes, the northern

blottings were carried out, as follows.

20 βg of each of the total RNA samples obtained from the three normal

exocervical tissues, the three primary cervical cancer tissues and the two cervical cancer cell lines as described in Example 1 was denatured and electrophoresized in a 1 % formaldehyde agarose gel, and then the resultant agarose gels were transferred to nylon

membranes (Boehringer-Mannheim, Germany), respectively. The nylon membranes

were then hybridized at 42 ^°C overnight with the ³²P-labeled random prime probes

using the full-length GIG46 and MIG20 cDNAs. The northern blotting procedure was

repeated twice; one is that the blots were quantitified using the densitometer and the

other is that the blots were hybridized with the β -actin probe to determine the total

amount of mRNA.

FIG. 77 shows the northern blotting result that the GIG46 gene is differentially

expressed in a normal exocervical tissue, a primary cervical cancer tissue and a cervical

cancer cell line, and FIG. 77 is a northern blotting result showing expression of β -actin.

In FIG. 77, Lanes 1 to 3 represent the normal exocervical tissue samples, Lanes 4 to 6

represent the cervical cancer tissue samples, Lane 7 represents the sample of the cervical

cancer cell line HeLa, and Lane 8 represents the sample of the cervical cancer cell line

CUMC-6. As shown in FIG. 77, it was revealed that the expression level of the GIG46

gene was highly detected all in the three samples of the normal exocervical tissue, but

its expression level was significantly lower in the three samples of the cervical cancer tissue than the normal tissue, and not detected in the two samples of the cervical cancer

cell lines.

FIG. 106 shows a northern blotting result that the GIG46 gene is differentially expressed in various normal tissues, and FIG. 106 shows a northern blotting result

obtained by hybridizing the same blot with β -actin probe. As shown in FIG. 106, a

dominant GIG46 mRNA transcript having a size of approximately 1.5 kb was

overexpressed in the normal tissues such as the uterus, the brain, the heart, the skeletal muscle, the large intestine, the thymus, the spleen, the kidney, the liver, the small

intestine, the placenta, the lungs and the peripheral blood leukocyte, and a transcript

having a size of approximately 2.0 kb was also expressed in addition to the 1.5 kb-GIG46 mRNA transcript.

FIG. 135 shows a northern blotting result that the GIG46 gene is differentially

expressed in various cancer cell lines, and a bottom of FIG. 135 shows a northern

in FIG. 135, the GIG46 gene was rarely expressed in the tissues such as the promyelocytic leukemia HL-60, the HeLa cervical cancer cell, the chronic myelocytic leukemia cell line K-562, the lymphoblastoid leukemia MOLT-4, the Burkitt's

melanoma cell. However, the 2.0 kb-mRNA transcrip proven to be expressed in the

normal tissues was not expressed in the cancer cell lines.

From such a result, it might be seen that the GIG46 gene of the present invention had the tumor suppresser function in the normal tissues such as the uterus, the brain, the

heart, the skeletal muscle, the large intestine, the thymus, the spleen, the kidney, the

liver, the small intestine, the placenta, the lungs and the peripheral blood leukocyte.

FIG. 81 shows the northern blotting result that the MIG20 gene is differentially expressed in a normal exocervical tissue, a primary cervical cancer tissue and a cervical

cancer cell line, and FIG. 81 is a northern blotting result showing expression of β -actin.

In FIG. 81, Lanes 1 to 3 represent the normal exocervical tissue samples, Lanes 4 to 6 represent the cervical cancer tissue samples, Lane 7 represents the sample of the cervical

cancer cell line HeLa, and Lane 8 represents the sample of the cervical cancer cell line CUMC-6. As shown in FIG. 81, it was revealed that the expression level of the

MIG20 gene was highly detected all in the three samples of the normal exocervical

tissue, but its expression level was significantly lower in the three samples of the

cervical cancer tissue than the normal tissue, and not detected in the two samples of the

cervical cancer cell lines.

FIG. 110 shows a northern blotting result that the MIG20 gene is differentially expressed in various normal tissues, and FIG. 110 shows a northern blotting result

obtained by hybridizing the same blot with β -actin probe. As shown in FIG. 110, a

dominant MIG20 mRNA transcript having a size of approximately 4.4 kb was

overexpressed in the normal tissues such as the heart, the skeletal muscle and the liver,

and transcripts having sizes of approximately 2.4 kb and 1.5 kb were also expressed in

addition to the 4.4 kb-MIG20 mRNA transcript.

FIG. 139 shows a northern blotting result that the MIG20 gene is differentially

expressed in various cancer cell lines, and a bottom of FIG. 139 shows a northern

in FIG. 139, the MIG20 gene was not expressed in the tissues such as the promyelocyte

leukemia HL-60, the HeLa cervical cancer cell, the chronic myelocytic leukemia cell line K-562, the lymphoblastoid leukemia MOLT-4, the Burkitt's lymphoma Raji, the

SW480 colon cancer cell, the A549 lung cancer cell and the G361 melanoma cell.

From such a result, it might be seen that the MIG2 gene of the present invention had the tumor suppresser function in the normal tissues such as the cervix, the heart, the skeletal

muscle and the liver.

3-5. MIG12 In order to assess an expression level of the MIGl 2 gene, the northern blotting

was carried out, as follows.

20 βg of each of the total RNA samples obtained from the three normal lung

tissues, the two primary lung cancer tissues, the two metastatic lung cancer tissues and

the lung cancer cell lines (A549 and NCI-H358) as described in Example 1 was

denatured and electrophoresized in a 1 % formaldehyde agarose gel, and then the

resultant agarose gel was transferred to a nylon membrane (Boehringer-Mannheim,

Germany). The nylon membrane was then hybridized at 42 °C overnight with the

³²P-labeled random prime probe prepared from the full-length MIG 12 cDNA using the

Rediprime II random prime labelling system (Amersham, United Kingdom). The

northern blotting procedure was repeated twice; one is that the blots were quantitified

using the densitometer and the other is that the blots were hybridized with the β -actin

probe to determine the total amount of mRNA.

FIG. 84 shows the northern blotting result that the MIGl 2 gene is differentially

expressed in a normal lung tissue, a primary lung cancer tissue, a metastatic lung cancer

tissue and a lung cancer cell line, and a bottom of FIG. 84 shows the northern blotting

84, it was revealed that the expression level of the MIG9 gene was highly detected all in

the three samples of the normal lung tissue, but slightly detected in the two samples of the lung cancer tissue, the two samples of the metastatic lung cancer tissue and the two

samples of the lung cancer cell line.

The northern blotting was carried out on the normal human multiple tissue (Clontech) and the human cancer cell line (Clontech). That is to say, the northern blotting was carried out by hybridizing blots, to which each of the total RNA samples

HeLa cervical cancer cell, a chronic myelocytic leukemia cell line K-562,

cell, an A549 lung cancer cell and a G361 melanoma cell.

FIG. 113 shows a northern blotting result that the MIG 12 gene is differentially

expressed in various normal tissues, and FIG. 113 shows a northern blotting result

obtained by hybridizing the same blot with β -actin probe. As shown in FIG. 113, a

dominant MIG12 mRNA transcript having a size of approximately 0.5 kb was overexpressed in the normal tissues such as the brain, the heart, the skeletal muscle, the colon, the thymus, the spleen, the kidney, the liver, the small intestine, the placenta, the

lung and the peripheral blood leukocyte. Transcripts having sizes of approximately 1.0

kb and 0.8 kb were also expressed in the normal tissues such as the heart, the muscles, the liver and the kidney in addition to the 0.5 kb-MIG12 mRNA transcript. FIG. 142 shows a northern blotting result that the MIG 12 gene is differentially expressed in various cancer cell lines, and a bottom of FIG. 142 shows a northern

in FIG. 142, the dominant 0.5 kb-MIG12 mRNA transcript expressed in the normal tissues was rarely expressed in the tissues such as the promyelocyte leukemia HL-60,

lymphoblastoid leukemia MOLT-4, the Burkitt's lymphoma Raji, the SW480 colon

it might be seen that the MIG 12 gene of the present invention had the tumor suppresser

function in the normal tissues such as the brain, the heart, the skeletal muscle, the colon,

the thymus, the spleen, the kidney, the liver, the small intestine, the placenta, the lung

and the peripheral blood leukocyte.

Example 4: Construction and Transfection of Expression Vector

4-1. GIG8. GIGlO. GIG13. GIG30. GIG32. GIG33, GIG34. GIG35, GIG38.

GIG39. GIG43. PIG49, PIG51. GIG44, GIG31

An expression vector containing each coding region of GIG and PIG genes was

constructed, as follows. Firstly, the full-length cDNA clones prepared in Example 2

were inserted into a eukaryotic expression vector pcDNA3.1 (Invitrogen, U.S.) to obtain

expression vectors pcDNA3.1/GIG8; pcDNA3.1/GIG10; pcDNA3.1/GIG13; pcDNA3.1/GIG30; pcDNA3.1/GIG32; pcDNA3.1/GIG33; pcDNA3.1/GIG34; pcDNA3.1/GIG35; pcDNA3.1/GIG38; pcDNA3.1/GIG39; pcDNA3.1/GIG43; pcDNA3.1/PIG49; pcDNA3.1/PIG51; pcDNA3.1/GIG44 and pcDNA3.1/GIG31, respectively. Each of the expression vectors was transfected into an MCF-7 breast cancer cell line using lipofectamine (Gibco BRL), and then incubated in a DMEM

medium containing 0.6 mg/iώ of G418 (Gibco) to select transfected cells. At this time,

the MCF-7 cell transfected with the expression vector pcDNA3.1 devoid of the GIG cDNA was used as the control group.

4-2. GIGl 5

An expression vector containing a coding region of the GIG 15 gene was

constructed, as follows. Firstly, the full-length GIG 15 cDNA clones prepared in

Example 2 was inserted into a eukaryotic expression vector pcDNA3.1 (Invitrogen,

U.S.) to obtain an expression vector pcDNA3.1/GIG15. The expression vector was

transfected into a K562 leukemia cell line using lipofectamine (Gibco BRL), and then

incubated in a DMEM medium containing 0.6 mg/ml of G418 (Gibco) to select

transfected cells. At this time, the K562 cell transfected with the expression vector

pcDNA3.1 devoid of the GIG cDNA was used as the control group.

4-3. GIG16, GIG24, GIG26, GIG29. GIG40. GIG42. PIG33. PIG35. PIG36.

PIG37

An expression vector containing each coding region of GIG and PIG genes was

constructed, as follows. Firstly, the full-length cDNA clones GIGl 6, GIG24, GIG26,

GIG29, GIG40, GIG42, PIG33, PIG35, PIG36 and PIG37 prepared in Example 2 were

inserted into a eukaryotic expression vector pcDNA3.1 (Invitrogen, U.S.) to obtain

expression vectors pcDNA3.1/GIG16; pcDNA3.1/GIG24; pcDNA3.1/GIG26;

pcDNA3.1/GIG29; pcDNA3.1/GIG40; pcDNA3.1/GIG42; pcDNA3.1/PIG33;

pcDNA3.1/PIG35; pcDNA3.1/PIG36; and pcDNA3.1/PIG37, respectively. Each of

the expression vectors was transfected into an HepG2 liver cancer cell line using lipofectamine (Gibco BRL), and then incubated in a DMEM medium containing 0.6

mg/M of G418 (Gibco) to select transfected cells. At this time, the HepG2 cell

transfected with the expression vector pcDNA3.1 devoid of the GIG cDNA was used as the control group.

4-4. GIG46. MIG20

An expression vector containing each coding region of the GIG and MIG genes

was constructed, as follows. Firstly, the full-length GIG46 cDNA clones prepared in

U.S.) to obtain expression vectors pcDNA3.1/GIG46; and pcDNA3.1/MIG20,

respectively. Each of the expression vectors was transfected into an HeLa cervical cancer cell line(ATCC CCL-2) using lipofectamine (Gibco BRL), and then incubated in

a DMEM medium containing 0.6 mg/mβ of G418 (Gibco) to select transfected cells.

At this time, the HeLa cell transfected with the expression vector pcDNA3.1 devoid of the GIG46 or MIG20 cDNA was used as the control group.

4-5. MIG12

An expression vector containing a coding region of the MIG 12 gene was

constructed, as follows. Firstly, the full-length MIG 12 cDNA clones prepared in

U.S.) to obtain an expression vector pcDNA3.1/MIG12. The expression vector was

transfected into an A549 lung cancer cell line using lipofectamine (Gibco BRL), and

then incubated in a DMEM medium containing 0.6 mg/mi of G418 (Gibco) to select

transfected cells. At this time, the A549 cell transfected with the expression vector

pcDNA3.1 devoid of the MIG 12 cDNA was used as the control group.

Example 5: Growth Curve of Breast Cancer Cell Transfected with GIG Gene 5-1. GIG8. GIGlO, GIG13, GIG30, GIG32, GIG33. GIG34, GIG35. GIG38, GIG39, GIG43. PIG49, PIG51. GIG44, GIG31

In order to examine effects of the GIG and PIG genes on growth of the breast

cancer cell, the wild-type MCF-7 cell; the MCF-7 breast cancer cells transfected

respectively with the vectors pcDNA3.1/GIG8; pcDNA3.1/GIG10; pcDNA3.1/GIG13;

pcDNA3.1/GIG30; pcDNA3.1/GIG32; pcDNA3.1/GIG33; pcDNA3.1/GIG34;

pcDNA3.1/GIG35; pcDNA3.1/GIG38; pcDNA3.1/GIG39; pcDNA3.1/GIG43; pcDNA3.1/PIG49; pcDNA3.1/PIG51; pcDNA3.1/GIG44 and pcDNA3.1/GIG31

prepared in Example 4; and the MCF-7 cell transfected only with the vector pcDNA3.1

were incubated to a cell density of 1 x 10⁵ cells/m# in a DMEM medium for 9 days,

respectively. The cells in the culture solutions were isolated from the flask they attach to by treatment with trypsin (Sigma), respectively, and then the survived cells were

counted on days 1, 3, 5, 7 and 9 according to a trypan blue dye exclusion (Freshney, LR. ,

Culture of Animal Cells, 2nd Ed. A.R. Liss, New York (1987)), respectively.

FIG. 146 is a diagram showing growth curves of the wild-type MCF-7 cell; the

MCF-7 breast cancer cell transfected with the vector pcDNA3.1/GIG8 prepared in

Example 4; and the MCF-7 cell transfected only with the expression vector pcDNA3.1.

As shown in FIG. 146, it was revealed that the MCF-7 breast cancer cell transfected with the vector pcDNA3.1/GIG8 exhibited a higher mortality than those of the MCF-7 cell transfected with the expression vector pcDNA3.1 and the wild-type MCF-7 cell.

After 9 days of incubation, only approximately 30 % of the MCF-7 breast cancer cell transfected with the vector pcDNA3.1/GIG8 was survived when compared to the

wild-type MCF-7 cell. From such a result, it might be seen that the GIG8 gene

suppressed the growth of the breast cancer cell. FIG. 147 is a diagram showing growth curves of the wild-type MCF-7 cell; the

MCF-7 breast cancer cell transfected with the vector pcDNA3.1/GIG10 prepared in

As shown in FIG. 147, it was revealed that the MCF-7 breast cancer cell transfected

with the vector pcDN A3.1 /GIG 10 exhibited a higher mortality than those of the MCF-7

cell transfected with the expression vector pcDNA3.1 and the wild-type MCF-7 cell.

After 9 days of incubation, only approximately 40 % of the MCF-7 breast cancer cell

transfected with the vector pcDNA3.1/GIG10 was survived when compared to the

wild-type MCF-7 cell. From such a result, it might be seen that the GIGlO gene suppressed the growth of the breast cancer cell.

FIG. 148 is a diagram showing growth curves of the wild-type MCF-7 cell; the

MCF-7 breast cancer cell transfected with the vector pcDNA3.1/GIG13 prepared in

As shown in FIG. 148, it was revealed that the MCF-7 breast cancer cell transfected with the vector pcDNA3.1/GIG13 exhibited a higher mortality than those of the MCF-7 cell transfected with the expression vector pcDNA3.1 and the wild-type MCF-7 cell. After 9 days of incubation, only approximately 30 % of the MCF-7 breast cancer cell transfected with the vector pcDNA3.1/GIG13 was survived when compared to the

wild-type MCF-7 cell. From such a result, it might be seen that the GIGl 3 gene

suppressed the growth of the breast cancer cell.

FIG. 154 is a diagram showing growth curves of the wild-type MCF-7 cell; the MCF-7 breast cancer cell transfected with the vector pcDNA3.1/GIG3O prepared in

Example 4; and the MCF-7 cell transfected only with the expression vector pcDNA3.1. As shown in FIG. 154, it was revealed that the MCF-7 breast cancer cell transfected

with the vector pcDNA3.1/GIG30 exhibited a higher mortality than those of the MCF-7

After 9 days of incubation, only approximately 30 % of the MCF-7 breast cancer cell transfected with the vector pcDNA3.1/GIG30 was survived when compared to the

wild-type MCF-7 cell. From such a result, it might be seen that the GIG30 gene suppressed the growth of the breast cancer cell.

FIG. 155 is a diagram showing growth curves of the wild-type MCF-7 cell; the

MCF-7 breast cancer cell transfected with the vector pcDNA3.1/GIG32 prepared in

As shown in FIG. 155, it was revealed that the MCF-7 breast cancer cell transfected

with the vector pcDNA3.1/GIG32 exhibited a higher mortality than those of the MCF-7 cell transfected with the expression vector pcDNA3.1 and the wild-type MCF-7 cell. After 9 days of incubation, only approximately 50 % of the MCF-7 breast cancer cell transfected with the vector pcDNA3.1/GIG32 was survived when compared to the

wild-type MCF-7 cell. From such a result, it might be seen that the GIG32 gene

suppressed the growth of the breast cancer cell.

FIG. 156 is a diagram showing growth curves of the wild-type MCF-7 cell; the

MCF-7 breast cancer cell transfected with the vector pcDNA3.1/GIG33 prepared in Example 4; and the MCF-7 cell transfected only with the expression vector pcDNA3.1. As shown in FIG. 156, it was revealed that the MCF-7 breast cancer cell transfected

with the vector pcDNA3.1/GIG33 exhibited a higher mortality than those of the MCF-7 cell transfected with the expression vector pcDNA3.1 and the wild-type MCF-7 cell. After 9 days of incubation, only approximately 70 % of the MCF-7 breast cancer cell

transfected with the vector pcDNA3.1/GIG33 was survived when compared to the

wild-type MCF-7 cell. From such a result, it might be seen that the GIG33 gene

suppressed the growth of the breast cancer cell.

FIG. 157 is a diagram showing growth curves of the wild-type MCF-7 cell; the

MCF-7 breast cancer cell transfected with the vector pcDNA3.1/GIG34 prepared in

As shown in FIG. 157, it was revealed that the MCF-7 breast cancer cell transfected

with the vector pcDNA3.1/GIG34 exhibited a higher mortality than those of the MCF-7

After 9 days of incubation, only approximately 80 % of the MCF-7 breast cancer cell

transfected with the vector pcDNA3.1/GIG34 was survived when compared to the

wild-type MCF-7 cell. From such a result, it might be seen that the GIG34 gene

suppressed the growth of the breast cancer cell. FIG. 158 is a diagram showing growth curves of the wild-type MCF-7 cell; the

MCF-7 breast cancer cell transfected with the vector pcDNA3.1/GIG35 prepared in

As shown in FIG. 158, it was revealed that the MCF-7 breast cancer cell transfected with the vector pcDNA3.1/GIG35 exhibited a higher mortality than those of the MCF-7

cell transfected with the expression vector pcDNA3.1 and the wild-type MCF-7 cell. After 9 days of incubation, only approximately 70 % of the MCF-7 breast cancer cell

transfected with the vector pcDNA3.1/GIG35 was survived when compared to the

wild-type MCF-7 cell. From such a result, it might be seen that the GIG35 gene suppressed the growth of the breast cancer cell.

FIG. 159 is a diagram showing growth curves of the wild-type MCF-7 cell; the

MCF-7 breast cancer cell transfected with the vector pcDNA3.1/GIG38 prepared in Example 4; and the MCF-7 cell transfected only with the expression vector pcDNA3.1.

As shown in FIG. 159, it was revealed that the MCF-7 breast cancer cell transfected

with the vector pcDNA3.1/GIG38 exhibited a higher mortality than those of the MCF-7

After 9 days of incubation, only approximately 60 % of the MCF-7 breast cancer cell

transfected with the vector pcDNA3.1/GIG38 was survived when compared to the

wild-type MCF-7 cell. From such a result, it might be seen that the GIG38 gene

suppressed the growth of the breast cancer cell.

FIG. 160 is a diagram showing growth curves of the wild-type MCF-7 cell; the

MCF-7 breast cancer cell transfected with the vector pcDNA3.1/GIG39 prepared in

As shown in FIG. 160, it was revealed that the MCF-7 breast cancer cell transfected

with the vector pcDNA3.1/GIG39 exhibited a higher mortality than those of the MCF-7 cell transfected with the expression vector pcDNA3.1 and the wild-type MCF-7 cell.

After 9 days of incubation, only approximately 40 % of the MCF-7 breast cancer cell transfected with the vector pcDNA3.1/GIG39 was survived when compared to the

wild-type MCF-7 cell. From such a result, it might be seen that the GIG39 gene suppressed the growth of the breast cancer cell.

FIG. 163 is a diagram showing growth curves of the wild-type MCF-7 cell; the MCF-7 breast cancer cell transfected with the vector pcDNA3.1/GIG43 prepared in Example 4; and the MCF-7 cell transfected only with the expression vector pcDNA3.1.

As shown in FIG. 163, it was revealed that the MCF-7 breast cancer cell transfected

with the vector pcDNA3.1/GIG43 exhibited a higher mortality than those of the MCF-7

transfected with the vector pcDNA3.1/GIG43 was survived when compared to the

wild-type MCF-7 cell. From such a result, it might be seen that the GIG43 gene

suppressed the growth of the breast cancer cell.

FIG. 169 is a diagram showing growth curves of the wild-type MCF-7 cell; the

MCF-7 breast cancer cell transfected with the vector pcDNA3.1/PIG49 prepared in

As shown in FIG. 169, it was revealed that the MCF-7 breast cancer cell transfected

with the vector pcDNA3.1/PIG49 exhibited a higher mortality than those of the MCF-7

cell transfected with the expression vector pcDNA3.1 and the wild-type MCF-7 cell. After 9 days of incubation, only approximately 60 % of the MCF-7 breast cancer cell

transfected with the vector pcDNA3.1/PIG49 was survived when compared to the wild-type MCF-7 cell. From such a result, it might be seen that the PIG49 gene

suppressed the growth of the breast cancer cell.

FIG. 170 is a diagram showing growth curves of the wild-type MCF-7 cell; the MCF-7 breast cancer cell transfected with the vector pcDNA3.1/PIG51 prepared in Example 4; and the MCF-7 cell transfected only with the expression vector pcDNA3.1.

As shown in FIG. 170, it was revealed that the MCF-7 breast cancer cell transfected

with the vector pcDNA3.1/PIG51 exhibited a higher mortality than those of the MCF-7 cell transfected with the expression vector pcDNA3.1 and the wild-type MCF-7 cell.

transfected with the vector pcDNA3.1/PIG51 was survived when compared to the

wild-type MCF-7 cell. From such a result, it might be seen that the PIG51 gene suppressed the growth of the breast cancer cell.

FIG. 173 is a diagram showing growth curves of the wild-type MCF-7 cell; the

MCF-7 breast cancer cell transfected with the vector pcDNA3.1/GIG44 prepared in

As shown in FIG. 173, it was revealed that the MCF-7 breast cancer cell transfected with the vector pcDN A3.1 /GIG44 exhibited a higher mortality than those of the MCF-7

transfected with the vector pcDNA3.1/GIG44 was survived when compared to the

wild-type MCF-7 cell. From such a result, it might be seen that the GIG44 gene

suppressed the growth of the breast cancer cell.

FIG. 174 is a diagram showing growth curves of the wild-type MCF-7 cell; the MCF-7 breast cancer cell transfected with the vector pcDNA3.1/GIG31 prepared in Example 4; and the MCF-7 cell transfected only with the expression vector pcDNA3.1.

As shown in FIG. 174, it was revealed that the MCF-7 breast cancer cell transfected

with the vector pcDN A3.1 /GIG31 exhibited a higher mortality than those of the MCF-7 cell transfected with the expression vector pcDNA3.1 and the wild-type MCF-7 cell.

After 9 days of incubation, only approximately 70 % of the MCF-7 breast cancer cell transfected with the vector pcDNA3.1/GIG31 was survived when compared to the wild-type MCF-7 cell. From such a result, it might be seen that the GIG31 gene

suppressed the growth of the breast cancer cell.

5-2. GIGl 5

In order to examine effects of the GIG gene on growth of the leukemia cell, the wild-type K562cell; the K562 leukemia cell transfected respectively by the vector

pcDNA3.1/GIG15 prepared in Example 4; and the K562 cell transfected only with the

vector pcDNA3.1 were incubated to a cell density of 1 x 10⁵ cells/mβ in a DMEM

medium for 9 days, respectively. The cells in the culture solutions were isolated from

the flask they attach to by treatment with trypsin (Sigma), respectively, and then the

survived cells were counted on days 1, 3, 5, 7 and 9 according to a trypan blue dye

exclusion (Freshney, I.R., Culture of Animal Cells, 2nd Ed. A.R. Liss, New York

(1987)), respectively.

FIG. 149 is a diagram showing growth curves of the wild-type K562 cell; the

K562 leukemia cell transfected with the vector pcDNA3.1/GIG15 prepared in Example

4; and the K562 cell transfected only with the expression vector pcDNA3.1. As shown in FIG. 149, it was revealed that the K562 cell transfected with the vector

pcDNA3.1/GIG15 exhibited a higher mortality than those of the K562 cell transfected with the expression vector pcDNA3.1 and the wild-type K562 cell. After 9 days of

incubation, only approximately 80 % of the K562 cell transfected with the vector pcDNA3.1/GIG15 was survived when compared to the wild-type K562 cell. From such a result, it might be seen that the GIGl 5 gene suppressed the growth of the breast cancer cell.

5-3. GIG16. GIG24. GIG26. GIG29. GIG40. GIG42. PIG33, PIG35. PIG36. PIG37

In order to examine effects of the GIG and PIG genes on growth of the liver

cancer cell, the wild-type HepG2 cell; the HepG2 liver cancer cells transfected

respectively by the vectors ρcDNA3.1/GIG16; pcDNA3.1/GIG24; ρcDNA3.1/GIG26;

pcDNA3.1/GIG29; pcDNA3.1/GIG40; pcDNA3.1/GIG42; pcDNA3.1/PIG33;

pcDNA3.1/PIG35; pcDNA3.1/PIG36; and pcDNA3.1/PIG37 prepared in Example 4;

and the HepG2 cell transfected only with the vector pcDNA3.1 were incubated to a cell

density of 1 x 10⁵ cells/mϋ. in a DMEM medium for 9 days, respectively. The cells in

the culture solutions were isolated from the flask they attach to by treatment with trypsin

(Sigma), respectively, and then the survived cells were counted on days 1, 3, 5, 7 and 9

according to a trypan blue dye exclusion (Freshney, I.R., Culture of Animal Cells, 2nd

Ed. A.R. Liss, New York (1987)), respectively.

FIG. 150 is a diagram showing growth curves of the wild-type HepG2 cell; the

HepG2 liver cancer cell transfected with the vector pcDNA3.1/GIG16 prepared in Example 4; and the HepG2 cell transfected only with the expression vector pcDNA3.1.

As shown in FIG. 150, it was revealed that the HepG2 liver cancer cell transfected with

the vector pcDNA3.1/GIG16 exhibited a higher mortality than those of the HepG2 cell

transfected with the expression vector pcDNA3.1 and the wild-type HepG2 cell. After 9 days of incubation, only approximately 70 % of the HepG2 liver cancer cell

transfected with the vector pcDNA3.1/GIG16 was survived when compared to the wild-type HepG2 cell. From such a result, it might be seen that the GIG 16 gene

suppressed the growth of the liver cancer cell.

FIG. 151 is a diagram showing growth curves of the wild-type HepG2 cell; the HepG2 liver cancer cell transfected with the vector pcDNA3.1/GIG24 prepared in

Example 4; and the HepG2 cell transfected only with the expression vector pcDNA3.1.

As shown in FIG. 151, it was revealed that the HepG2 liver cancer cell transfected with the vector pcDNA3.1/GIG24 exhibited a higher mortality than those of the HepG2 cell

transfected with the expression vector pcDNA3.1 and the wild-type HepG2 cell. After

9 days of incubation, only approximately 60 % of the HepG2 liver cancer cell

transfected with the vector pcDNA3.1/GIG24 was survived when compared to the

wild-type HepG2 cell. From such a result, it might be seen that the GIG24 gene

suppressed the growth of the liver cancer cell.

FIG. 152 is a diagram showing growth curves of the wild-type HepG2 cell; the

HepG2 liver cancer cell transfected with the vector pcDNA3.1/GIG26 prepared in

As shown in FIG. 152, it was revealed that the HepG2 liver cancer cell transfected with

the vector pcDNA3.1/GIG26 exhibited a higher mortality than those of the HepG2 cell

transfected with the expression vector pcDNA3.1 and the wild-type HepG2 cell. After 9 days of incubation, only approximately 50 % of the HepG2 liver cancer cell

transfected with the vector pcDNA3.1/GIG26 was survived when compared to the

wild-type HepG2 cell. From such a result, it might be seen that the GIG26 gene suppressed the growth of the liver cancer cell.

FIG. 153 is a diagram showing growth curves of the wild-type HepG2 cell; the

HepG2 liver cancer cell transfected with the vector pcDNA3.1/GIG29 prepared in Example 4; and the HepG2 cell transfected only with the expression vector pcDNA3.1.

As shown in FIG. 153, it was revealed that the HepG2 liver cancer cell transfected with the vector pcDNA3.1/GIG29 exhibited a higher mortality than those of the HepG2 cell

9 days of incubation, only approximately 70 % of the HepG2 liver cancer cell

transfected with the vector pcDNA3.1/GIG29 was survived when compared to the

wild-type HepG2 cell. From such a result, it might be seen that the GIG29 gene

suppressed the growth of the liver cancer cell.

FIG. 161 is a diagram showing growth curves of the wild-type HepG2 cell; the HepG2 liver cancer cell transfected with the vector pcDNA3.1/GIG40 prepared in

As shown in FIG. 161, it was revealed that the HepG2 liver cancer cell transfected with

the vector pcDNA3.1/GIG40 exhibited a higher mortality than those of the HepG2 cell

transfected with the expression vector pcDNA3.1 and the wild-type HepG2 cell. After 9 days of incubation, only approximately 80 % of the HepG2 liver cancer cell

transfected with the vector pcDNA3.1/GIG40 was survived when compared to the

wild-type HepG2 cell. From such a result, it might be seen that the GIG40 gene

suppressed the growth of the liver cancer cell.

FIG. 162 is a diagram showing growth curves of the wild-type HepG2 cell; the HepG2 liver cancer cell transfected with the vector pcDNA3.1/GIG42 prepared in

Example 4; and the HepG2 cell transfected only with the expression vector pcDNA3.1. As shown in FIG. 162, it was revealed that the HepG2 liver cancer cell transfected with the vector pcDNA3.1/GIG42 exhibited a higher mortality than those of the HepG2 cell

9 days of incubation, only approximately 60 % of the HepG2 liver cancer cell transfected with the vector pcDNA3.1/GIG42 was survived when compared to the

wild-type HepG2 cell. From such a result, it might be seen that the GIG42 gene

suppressed the growth of the liver cancer cell.

FIG. 165 is a diagram showing growth curves of the wild-type HepG2 cell; the

HepG2 liver cancer cell transfected with the vector pcDNA3.1/PIG33 prepared in

As shown in FIG. 165, it was revealed that the HepG2 liver cancer cell transfected with

the vector pcDNA3.1/PIG33 exhibited a higher mortality than those of the HepG2 cell transfected with the expression vector pcDNA3.1 and the wild-type HepG2 cell. After

9 days of incubation, only approximately 60 % of the HepG2 liver cancer cell

transfected with the vector pcDNA3.1/PIG33 was survived when compared to the

wild-type HepG2 cell. From such a result, it might be seen that the PIG33 gene

suppressed the growth of the liver cancer cell.

FIG. 166 is a diagram showing growth curves of the wild-type HepG2 cell; the HepG2 liver cancer cell transfected with the vector pcDNA3.1/PIG35 prepared in Example 4; and the HepG2 cell transfected only with the expression vector pcDNA3.1.

As shown in FIG. 166, it was revealed that the HepG2 liver cancer cell transfected with the vector pcDNA3.1/PIG35 exhibited a higher mortality than those of the HepG2 cell

9 days of incubation, only approximately 70 % of the HepG2 liver cancer cell transfected with the vector pcDNA3.1/PIG35 was survived when compared to the

wild-type HepG2 cell. From such a result, it might be seen that the PIG35 gene

suppressed the growth of the liver cancer cell. FIG. 167 is a diagram showing growth curves of the wild-type HepG2 cell; the

HepG2 liver cancer cell transfected with the vector pcDNA3.1/PIG36 prepared in

As shown in FIG. 167, it was revealed that the HepG2 liver cancer cell transfected with the vector pcDNA3.1/PIG36 exhibited a higher mortality than those of the HepG2 cell

9 days of incubation, only approximately 60 % of the HepG2 liver cancer cell

transfected with the vector pcDNA3.1/PIG36 was survived when compared to the

wild-type HepG2 cell. From such a result, it might be seen that the PIG36 gene suppressed the growth of the liver cancer cell.

FIG. 172 is a diagram showing growth curves of the wild-type HepG2 cell; the

HepG2 liver cancer cell transfected with the vector pcDNA3.1/PIG37 prepared in

As shown in FIG. 172, it was revealed that the HepG2 liver cancer cell transfected with

the vector pcDNA3.1/PIG37 exhibited a higher mortality than those of the HepG2 cell

9 days of incubation, only approximately 70 % of the HepG2 liver cancer cell transfected with the vector pcDNA3.1/PIG37 was survived when compared to the wild-type HepG2 cell. From such a result, it might be seen that the PIG37 gene

suppressed the growth of the liver cancer cell.

5-4. GIG46, MIG20

In order to determine effects of the GIG and MIG genes on growth of the cervical cancer cell, the normal HeLa cell, the HeLa cervical cancer cell transfected with the GIG46 gene prepared in Example 4, and the HeLa cell transfected only with the

vector pcDNA3.1 (Invitrogen) were incubated to a cell density of 1 x 10⁵ cells/in^ in a

DMEM medium for 9 days, respectively. The cells in the culture solutions were

isolated from the flask they attach to by treatment with trypsin (Sigma), and then the

exclusion (Freshney, LR. , Culture of Animal Cells, 2nd Ed. A.R. Liss, New York (1987)).

FIG. 164 is a diagram showing growth curves of the normal HeLa cell; the HeLa

cervical cancer cell transfected with the GIG46 gene prepared in Example 4; and the

HeLa cell transfected only with the expression vector pcDNA3.1. As shown in FIG.

164, it was revealed that the HeLa cervical cancer cell transfected with the GIG46 gene

exhibited a higher mortality when compared to those of the HeLa cell transfected with the expression vector pcDNA3.1 and the normal HeLa cell. After 9 days of incubation,

only 80 % of the HeLa cervical cancer cell transfected with the GIG46 gene was survived when compared to the normal HeLa cell. From such a result, it might be seen

that the GIG46 gene suppressed growth of the cervical cancer cell.

FIG. 168 is a diagram showing growth curves of the normal HeLa cell; the HeLa cervical cancer cell transfected with the MIG20 gene prepared in Example 4; and the

HeLa cell transfected only with the expression vector pcDNA3.1. As shown in FIG. 168, it was revealed that the HeLa cervical cancer cell transfected with the MIG20 gene exhibited a higher mortality when compared to those of the HeLa cell transfected with

the expression vector pcDNA3.1 and the normal HeLa cell. After 9 days of incubation,

only approximately 60 % of the HeLa cervical cancer cell transfected with the MIG20 gene was survived when compared to the normal HeLa cell. From such a result, it

might be seen that the MIG20 gene suppressed growth of the cervical cancer cell. 5-5. MIG12

In order to determine an effect of the MIG 12 gene on growth of the lung cancer

cell, the wild-type A549 cell; the A549 lung cancer cell transfected with the vector

pcDNA3.1/MIG12 prepared in Example 4; and the A549 cell transfected only with the

vector pcDNA3.1 were incubated at a cell density of 1 x 10⁵ cellsM in a DMEM

medium for 9 days, respectively. The cells in the culture solutions were isolated from the flask they attach to by treatment with trypsin (Sigma), and then the survived cells were counted on days 1, 3, 5, 7 and 9 according to a trypan blue dye exclusion (Freshney,

I.R., Culture of Animal Cells, 2nd Ed. A.R. Liss, New York (1987)).

FIG. 171 is a diagram showing growth curves of the wild-type A549 cell; the

A549 lung cancer cell transfected by the vector pcDNA3.1/MIG12 prepared in Example

4; and the A549 cell transfected only by the expression vector pcDNA3.1. As shown

in FIG. 171, it was revealed that the A549 lung cancer cell transfected by the vector pcDNA3.1/MIG12 exhibited a higher mortality when compared to those of the A549 cell transfected by the expression vector pcDNA3.1 and the wild-type A549 cell. After 9 days of incubation, only approximately 70 % of the A549 lung cancer cell transfected

by the vector pcDNA3.1/MIG12 was survived when compared to the wild-type A549

cell. From such a result, it might be seen that the MIG12 gene suppressed growth of the lung cancer cell.

INDUSTRIAL APPLICABILITY The GIG, PIG or MIG gene of the present invention may be effectively used for

diagnosing, preventing and treating human cancers.

Claims

What is claimed is;

1. A human cancer suppressor protein having an amino acid sequence

selected from the group consisting of SEQ ID NO: 2; 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; SEQ ID NO: 62; SEQ ID NO: 66; SEQ ID NO: 70;

SEQ ID NO: 74; SEQ ID NO: 78; SEQ ID NO: 82; SEQ ID NO: 86; SEQ ID NO: 90;

SEQ ID NO: 94; SEQ ID NO: 98; SEQ ID NO: 102; SEQ ID NO: 106; SEQ ID NO:

110 and SEQ ID NO: 114.

2. The human cancer suppressor protein according to claim 1, wherein the

cancer is a cancer of a tissue selected from the group consisting of normal breast, brain, heart, muscles, large intestine, thymus, spleen, kidney, liver, small intestine, placenta,

lungs and peripheral blood.

3. A human cancer suppressor gene encoding the protooncoprotein defined in claim 1, the human cancer suppressor gene being set forth in a DNA sequence

selected from the group consisting of 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; SEQ ID NO: 61; SEQ ID NO: 65; SEQ ID NO: 69; SEQ ID NO: 73; SEQ ID NO: 77; SEQ ID NO: 81; SEQ ID NO: 85; SEQ ID NO: 89; SEQ ID NO: 93; SEQ ID NO: 97; SEQ ID NO: 101; SEQ ID NO: 105; SEQ ID NO:

109; and SEQ ID NO: 113.

4. The human cancer suppressor gene according to claim 3, wherein the

cancer is a cancer of a tissue selected from the group consisting of normal breast, brain,

heart, muscles, large intestine, thymus, spleen, kidney, liver, small intestine, placenta,

lungs and peripheral blood.