JPH08107797A - Dna coding protein related to repair of mismatch of human dna - Google Patents

Abstract

PURPOSE: To obtain the subject new DNA existing in the seventh chromosome long arm, coding a human DNA mismatch repair-relating protein having a specific base sequence, used for the analysis or diagnosis of genes with the gene, etc., and enabling the risk diagnosis or early discovery of cancers, etc. CONSTITUTION: A DNA coding the human DNA mismatch repair-relating protein consisting of six kinds of new DNAs comprising sequences each of which contains the sequence of the formula, or a DNA containing the new DNAs, and a gene coding the human DNA mismatch repair-relating protein. The DNA is used as a primer or probe in a gene-analyzing or diagnosing method utilizing the gene, etc., used for the isolation, immobilization, etc., of the gene coding a protein related to the DNA mismatch-repairing mechanisms of man or a higher organism, and further, used for the risk diagnosis, prophylactic treatment, early discovery, or progress observation of cancers, etc. The DNA is obtained by cloning the cDNA library of a human fetal brain on the basis of a partial sequence well stored in the DNA mismatch-repairing protein gene of a bacterium or yeast.

Description

Detailed Description of the Invention

[0001]

TECHNICAL FIELD The present invention relates to a gene DNA encoding a novel human DNA mismatch repair-related protein and said D
A gene analysis method using a protein encoded by NA or the DNA is used in the fields of medicine, diagnosis and the like.

[0002]

2. Description of the Related Art In DNA of a cell, a base pair that cannot be partially paired with a double strand (mismatch) is generated by the introduction of an incorrect base during its replication or the occurrence of a mutation by various mutagens. Although it may occur, it has long been known that bacteria and yeast have a mechanism for repairing this. For example, when a male strain of Salmonella having a mutation in the DNA mismatch repair mechanism (mutS, mutL) is mated with a male strain of Escherichia coli, the heterologous group is 1000 times as much as that of a wild type female strain having no mutation. It has been reported that replacement will occur. (Christiane et al.,
(Nature 342, 396-401, 1989) It has been studied that the DNA mismatch repair mechanism has an interfering effect even in the heterologous transformation of Streptococcus pneumoniae and Streptococcus, and its repair gene is named after Hex recombination.
They are named A and HexB. (Claverys JP et a
l., Microbial Rev. 50, 133-165, 1986) The best studied to date is Mu in E. coli.
It is a DNA mismatch repair mechanism (Mut system) by tH, MutL, and MutS proteins, and it has been proved that these proteins are necessary for mismatch repair by in vitro reconstitution experiments. (Modrich P., J. Bio
L. Chem. 264, 6597-6600, 1989), (Modrich P., Annu.
Rev. Genet. 25, 229-253, 1991), (Lahue RS et a
L., Science 245, 160-164, 1989), (Cooper DL et
al., J. Biol. Chem. 268, 11823-11829, 1993) Yeast also has similar DNA mismatch repair proteins, which are genetically well conserved and correspond to MutL and MutS of Escherichia coli. Proteins PMS1, MLH
1, MSH2, etc. have been discovered. This MSH2, P
It has also been clarified that in mutants of genes encoding the MS1 and MLH1 proteins, repair is disordered and abnormalities in repeated DNA sequences due to mismatches frequently occur. (Kramer W. et al., J. Bacteriol. 171, 5339-5346,
1989), (Strand M. et al., Nature 365, 274-276, 1
993), (Reeman RAG et al., Genetics 132, 963-97
3, & 975-985, 1992) Several other DNA mismatch repair proteins have been discovered in bacteria and yeast, and it is considered that there are many repair systems corresponding to the types of DNA mismatch. . Moreover, those proteins are, for example, Mut proteins.
L and HexB, MutS and HexA, or HexB
As it was pointed out that MutL and MutL and PMS1 have characteristic structures similar to each other, it is predicted that they are derived from a common ancestor and that there are proteins that are closely related to the living world. There is. (Harber LT et al.,
J. Bacteriol. 170, 197-202, 1988), (Priebe S. et a
L., J. Bacteriol. 170, 190-196, 1988), (Prudomme M.
et al., J. Bacteriol. 171, 5332-5338, 1989).

[0003] Recently, it has become extremely interesting to find that, in various human cancers, DNA repeat sequence abnormalities similar to those resulting from malfunction of DNA mismatch repair occur one after another. (Ionov Y. et a
L., Nature 363, 558-561, 1993), (Thiobodeau SN e
t al., Science 260, 816-819, 1993), (Han HJ et a
l., Cancer 53, 5087-5089, 1993) and further, a genetic study of human hereditary non-polyposis colorectal cancer (HNPCC) identified a gene encoding the human DNA mismatch repair protein MSH2 followed by MLH1. It was shown that these abnormalities are deeply involved in the cause of HNPCC. (Fishel R. et al.,
Cell 75, 1027-1038, 1993), (Leach F. et al., Cell
75, 1215-1225, 1993), (Parsons R. et al., Cell 7
5, 1227-1236, 1993), (Marx J., Science 262, 1645,
1993), (Palombo F. et al., Nature 367, 417, 199.
4), (Bronner CE et al., Nature 368, 258-261, 199.
4), (Papadopoulos N. et al., Science 263, 1625-162
9, 1994), (Robert F., Science 263, 1559-1560, 199.
4), (Bodmer W. et al., Nature Genetics 6, 217-21
9,1994) In addition, MSH2, M found in this HNPCC study
Novel human DNA mismatch repair proteins PMS1 and PMS2 having a similar characteristic sequence from the LH1 DNA sequence
The gene coding for was also identified, and it was shown that these were mutated in HNPCC patients. (Nicolai
des NC et al., Nature 371, 75-80, 1994)

As described above, humans have a plurality of DNA mismatch repair mechanisms as in bacteria and yeast, and a large number of DNA mismatch repair proteins constitute them to repair various DNA mismatches. That the human D
It has been shown that mutations in the gene encoding the NA mismatch repair-related protein are closely associated with disease. Disorders of this mechanism result in abnormalities of various genes,
It is considered to be fundamentally related to the causes of many diseases caused by genetic abnormalities including genetic diseases as well as cancer. Therefore, it is necessary to newly isolate and identify a gene encoding an undiscovered DNA mismatch repair-related protein, particularly in human, to clarify the sequence, or, in addition, these DNAs.
Further elucidation of the whole picture of the DNA mismatch repair mechanism by the group of mismatch repair proteins and its abnormality is expected in particular in the medical field. Besides this,
It is also expected to promote cross-species genetic recombination by inhibiting the DNA mismatch repair mechanism in higher organism cells, and to eliminate the difficulty of preparing pathological model animals.
Isolation and identification of DNA mismatch repair proteins of other higher organisms or genes encoding them are also expected.
(Hinata Saito, Bioscience and Industry, 15
1, 993-994, 1993)

[0005]

DISCLOSURE OF THE INVENTION The present invention provides a gene encoding a novel protein involved in DNA mismatch repair,
It is another object of the present invention to provide a method for cloning an unknown DNA mismatch repair protein gene, a method for testing cancer, a genetic disease, a method for diagnosing the disease, etc.

[0006]

Amino acid sequences of DNA mismatch repair proteins, which have been widely found in the living world so far, are similar to each other, but the specific similar sequences are spread throughout the entire protein. Rather than being partially concentrated. For example, yeast PMS1 and bacterial H
ExB has a homology of 13% as a whole, but has a homology of 33% when compared in the region of amino acids 32 to 190 of the N-terminal side of yeast PMS1. Furthermore, the positions of individual homologous amino acids exist in several clusters (groups) within this concentrated region. (Kramer W.
et al., J. Bacteriol. 171, 5339-5346, 1989) The inventors designed a plurality of combinations of DNA sequences reverse-translated from homologous sequence clusters in the amino acid sequences of these yeast and bacterial DNA mismatch repair proteins. RNA synthesized from human tissues using these as primers
RT-PCR method (Frohman MA et al., P
Roc. Natl. Acad. Sci. USA 85, 8998-9002, 1988) was used to amplify the gene DNA sequence encoding a DNA mismatch repair protein analog in humans. further,
Using the PCR clone thus obtained as a probe, a cDNA library prepared from human fetal brain was screened and the nucleotide sequence was determined. As a result, 6 types of cDNA were obtained. It was confirmed that the amino acid sequences of the proteins encoded by these cDNAs all show a high degree of similarity to human PMS2 and are novel human DNA mismatch repair-related proteins that have never been reported.

Furthermore, the inventors have found that these 6 cDNAs are
By screening a cosmid library using a mixture of clones as a probe, 4 of these c
A total of 26 cosmid clones including those corresponding to the DNA clones were obtained. As a result of chromosome mapping by the FISH (fluorescent in situ hybridization) method using the cosmid clone as a probe, it was confirmed that all 26 cosmid clones were located on the long arm of chromosome 7. That is, the position on the chromosome of the gene of the present invention,
The position of the related human DNA mismatch repair protein gene on the chromosome, specifically, human MSH2 / 2 short arm, human MLH1 / 3 short arm, human PMS1 / 2
Chromosome long arm, human PMS 2/7 short arm, (Lindb
lom A. et al., Nature Genet. 5, 279-282, 1993) (Ni
colaides NC et al., Nature 371, 75-80, 1994).

The present invention uses, for example, all or part of the DNA of the present gene as a primer or a probe to obtain a gene encoding at least a new protein related to the DNA mismatch repair mechanism of humans or higher organisms. It is extremely important in that it can provide methods and materials for dissociation and identification, and can also provide solutions for cancer risk diagnosis, preventive measures, early detection, follow-up, treatment policy decision, prognosis estimation, etc. is there. That is, the present invention provides (1) SEQ ID NOS: 1, 2, 3, 4,
DNA consisting of a part or all of the gene DNA represented by 5 or 6, (2) SEQ ID NOS: 1, 2,
A protein comprising all or part of the protein encoded by the gene DNA represented by 3, 4, 5 or 6;
(3) DN of SEQ ID NO: 1, 2, 3, 4, 5 or 6
A plasmid containing all or part of A, a transformant carrying the plasmid, and a method for producing the protein using the transformant, (4) SEQ ID NOS: 1, 2, 3,
An antibody that binds to a protein containing all or part of the protein encoded by the DNA represented by 4, 5 or 6, (5) of the DNA sequence represented by SEQ ID NO: 1, 2, 3, 4, 5 or 6 It consists of a primer containing a part, a probe or a marker, and a gene analysis method characterized by using them. The present invention will be described in detail below.

(1) Structure of Gene DNA and Protein Encoded by It It was confirmed that this gene DNA is a novel DNA sequence of SEQ ID NOs: 1, 2, 3, 4, 5, and 6. DNA mismatch repair-related proteins that exist widely in the living world
The sequence can be divided into several strains. For example, MSH2 and MLH1, which are considered to play a major role among them, are arranged as so-called phylogenetic trees showing the interrelationship between species and their evolutionary divergence with respect to their analogues. (Fishel R. et a
L., Cell 75, 1027-1038, 1993), (Bronner CE et a
L., Nature 368, 258-261, 1994). The present inventors have investigated the major DNA mismatch repair-related proteins and the protein encoded by this gene DNA by the method of Higgins (Higgins DG et al., Comput. Applic. Biosci.
8, 189-191, 1992) based on the computer analysis software DNASIS-Mac Ver.3.00 (Hitachi Software Engineering Co., Ltd.), which was developed based on the sequence comparison (Higgins method multiple alignment). As a result, it was found that all the proteins encoded by this gene DNA were derived from a common ancestor with these major DNA mismatch repair-related proteins, and among them, the DNA mismatch repair-related proteins of the line closest to PMS2. (FIG. 1) (FIG. 2) (Numerical values in the figure represent the concordance rate.) The inventors have respectively identified the proteins encoded by the DNAs represented by SEQ ID NOs: 1, 2, 3, 4, 5 and 6. Human PMS
It was named 3, 4, 5, 6, 7 and 8. That is, the present invention has disclosed herein a base sequence of a gene encoding a group of new strains of DNA mismatch repair-related proteins. This means that at least, for example, as in the case of the means for the present invention, a new material for cloning a further undiscovered DNA mismatch repair-related protein gene using a part of the nucleotide sequence of the gene as a probe or as a primer. Is meant to be provided.

The DNA of the present invention can be used for gene cloning, gene analysis and diagnosis by using a part of the DNA as a primer or a probe.
The term "part" means that the oligonucleotide used as a primer or probe consists of at least about 6 corresponding nucleotide sequences based on the DNA sequence of the present invention, preferably at least about 8 nucleotide sequences, and more preferably About 1
It means a corresponding polynucleotide consisting of 0 to 12, more preferably about 15 to 25 nucleotide sequences. The protein encoded by the DNA can be used as a reagent for research and diagnostics using the antibody by using all or part of the protein as an epitope.
The "epitope" means an antigenic determinant of a polypeptide and is generally known to be composed of at least 5 amino acids, and a polypeptide composed of 6 amino acids to bind to an antibody (Published Patent Publication). 60-500684). The part of the protein means at least about 3 contiguous sequences based on the amino acid sequence of the protein encoded by the DNA of the present invention.
~ 5 amino acids, preferably at least about 8-10 amino acids, more preferably at least about 11-20.
It means a polypeptide consisting of 4 amino acids. It goes without saying that even a polypeptide consisting of about 20 or more amino acids can be used. Further, in the DNA and the part thereof, even if those sequences have one or more bases converted, deleted or inserted, the same effect is exerted for research, diagnosis and the like. Those are also included in the present invention. In the case of the protein encoded by the DNA of the present invention, the term "consisting of a part" has the same meaning as in the case of the above DNA.

(2) Recombinant expression vector and transformant thereof A transformant can be obtained by incorporating the gene DNA of the present invention or a fragment thereof into an appropriate vector and transferring the vector into an appropriate host cell. . By culturing this in a conventional manner, the protein encoded by the DNA of the present invention can be produced in large quantities from the culture. More specifically, the DNA or its fragment can be recombined by a known method using a restriction enzyme and DNA ligase downstream of the promoter of a vector suitable for its expression to prepare a recombinant expression vector. Vectors that can be used include, for example, the plasmid pRB322, pU derived from E. coli.
C18, Bacillus subtilis derived plasmid pUB110, yeast derived plasmid pRB15, bacteriophage λg
Examples of the vector include t10, λgt11, SV40 and the like, but the vector is not particularly limited as long as it can be replicated and amplified in the host. The promoter and terminator are not particularly limited as long as they correspond to the host used for expressing the DNA base sequence, and suitable combinations are possible depending on the host. The DNA used is SEQ ID NO: 1,
The base sequence is not limited to the base sequences described in 2, 3, 4, 5 or 6, and bases in which a part of the base sequence is substituted, deleted, inserted, or a combination thereof, regardless of intentional purpose. It may be a DNA having a sequence. Further, it may be one synthesized by chemical synthesis.

The recombinant expression vector thus obtained was obtained by the competent cell method (J. Mol. Biol., 53, 154, 197).
0), protoplast method (Proc. Natl. Acad. Sci. USA,
75, 1929, 1978), calcium phosphate method (Science, 22
1, 551, 1983), in vitro packaging method (Proc. N
Atl, Acad. Sci. USA, 72, 581, 1975), virus vector method (Cell, 37, 1053, 1984) and the like are introduced into a host to prepare a transformant. As a host, Escherichia coli, Bacillus subtilis, yeast, animal cells and the like are used, and the obtained transformant is cultured in an appropriate medium according to the host. Cultivation is usually carried out in the range of 20 ° C to 45 ° C and pH 5 to 8,
Aeration and agitation are performed as necessary. Separation / purification of the protein from the culture may be carried out by appropriately combining known separation / purification methods. Known methods include salting out, solvent precipitation, dialysis gel filtration, electrophoresis, ion exchange chromatography, affinity chromatography, reverse phase high performance liquid chromatography and the like.

(3) Preparation of antibody The antibody can be prepared by a conventional method using all or part of the protein encoded by the DNA of the present invention as an antigen. For example, a polyclonal antibody is subcutaneously, intramuscularly, intraperitoneally, or intravenously inoculated into animals such as mice, guinea pigs, rabbits, etc. multiple times to sufficiently immunize, and then blood is collected from such animals.
It is prepared by separating serum. In addition, a commercially available adjuvant can also be used. Monoclonal antibody, for example, after preparing a hybridoma obtained by cell fusion of splenocytes of mice immunized with the protein and commercially available mouse myeloma cells,
It can be prepared from the hybridoma culture supernatant or the ascites of the hybridoma-administered mouse. The protein used as an antigen does not necessarily have to have the entire structure of the protein encoded by the DNA of the present invention, but a peptide having a partial structure,
It may be a mutant, derivative, or fusion peptide with another peptide, and the preparation method may be a biological method or a chemical synthesis method. These antibodies enable identification and quantification of the protein in human biological samples and can be used as cancer diagnostic reagents. The immunological measurement method of the protein may be based on a known method, for example, a fluorescent antibody method, a passive agglutination reaction method, an enzyme antibody method or the like.

(4) Genetic Analysis of Biological Sample As a biological sample to be genetically analyzed, human normal tissue, various human cancer tissues, pathological tissues, human blood, body fluid, secretory fluid and the like can be used. Extraction / preparation of DNA is performed, for example, by the method of (Sato T., et al. Cancer Res., 50, 7184, 1990). Presence or absence of mutation of the gene by using the restriction enzyme fragment of the gene DNA provided by the present invention as a probe or appropriately selecting a nucleotide sequence at an appropriate position from the gene DNA and using the synthetic oligonucleotide as a primer. Can be analyzed. Also,
Abnormalities such as insertions and deletions in the gene in the sample can also be detected by these analyzes. As the site of the base sequence to be selected, any part of the gene can be selected. Needless to say, it is also possible to use an artificially modified base sequence to be used, whereby the corresponding gene mutation can be detected.

As a method of analysis, for example, a partial sequence is amplified by PCR with primers of two kinds of selected sequences, and the base sequence of the amplification product is directly analyzed, or this amplification product is subjected to plasmid analysis in the same manner as described above. , The host cell is transformed and cultured, and the nucleotide sequence of the obtained clone can be analyzed. Alternatively, LigaseCh
ain Reaction method (Wu et al., Genomics, 4, 560-569, 1
989), and a mutant sequence-specific PCR method (Ruano and Ki
dd, Nucleic Acid Research, 17, 8392, 1989), (CR
Newton et al., Nucleic Acid Research, 17, 2503-251
7, 1989), it is possible to directly detect the presence or absence of a specific mutation of the gene in the sample. Similarly, a selected DNA sequence or RNA derived therefrom
SSCP method, RNase-Prot using probe containing sequence
The point mutation can be detected by the ection method.
Further, by using these probes, it is possible to detect the mutation of the gene in the sample by the Southern hybridization method and the abnormality in the expression level of the gene in the sample by the Northern hybridization method.

[0016]

INDUSTRIAL APPLICABILITY According to the present invention, at least all or part of the gene DNA thereof is used as a primer or a probe to obtain a gene encoding a newer protein related to the DNA mismatch repair mechanism of humans or higher organisms. Methods and materials for separating and identifying were provided.
In addition, cancer risk diagnosis, preventive measures, early detection,
It is extremely important in that it can also provide methods and materials for genetic diagnosis / testing or antibody diagnosis / testing as a solution to follow-up, determination of treatment policy, prognosis estimation, and the like.

[0017]

The present invention will be described in detail and specifically with reference to the following examples, but the present invention is not limited to these examples.

(Example 1) RT-PCR with degenerate primers Known method from human gastric tissue excised for medical reasons
(Nakamura Y. et al., Gene 28, 263-270, 1984)
Otal RNA was extracted and purified. RT-PCR was performed using 10 μg of this RNA and a single-stranded cDNA obtained by a reverse transcriptase reaction as a template. The primers used were degenerate primers (Compton T., PCR designed based on the respective amino acid sequences selected from two homologous clusters of the amino acid sequences of the yeast PMS1, Escherichia coli MutL, and pneumococcal HexB proteins. Experimental manual, HBC Publishing Bureau, 35-40, 1991). The selected amino acid sequences and the sequences of degenerate primers based on them are shown below. The amino acid sequence is represented by the one-letter code. In addition, a BamHI cleavage site sequence for vector construction is added to the 5'end of each primer. (Sense strand primer) Amino acid sequence; (V / A) VKELV (E / D) DNA sequence; 5'-CTGGGATCCG (C / T)
(A / G / C / T) GT (A / G / C / T) AA (A /
G) GA (A / G) (C / T) T (A / G / C / T) G
T (A / G / C / T) GA-3 ′ (antisense strand primer) amino acid sequence; LGFRGEA DNA sequence; 3′-AA (T / C / G / A) CC (T /
C / G / A) AA (G / A) (T / G) C (T / C / G
/ A) CC (T / C / G / A) CT (C / T) CGCC
TAGGCTC-5 ′ PCR was performed according to a known method (Horii A. et al., Hum. Mo.
l. Genet. 2, 283-287, 1993), 94 ° C for 30 seconds, 45 ° C for 2 minutes,
72 ℃ 3 minutes cycle 8 times, 94 ℃ 30 seconds, 55 ℃ 30 seconds, 72
A cycle of 30 seconds at 40 ° C was repeated 40 times. The PCR reaction solution was subjected to 10% acrylamide gel electrophoresis and the yeast PMS
In the case of 1, a band having the same length as the expected product was extracted and cloned into pBluescript II SK (-) vector (Stratagene) using the BamHI site. As a result of investigating the nucleotide sequence of each clone, clone No. 1 showing clear homology with the sequence of yeast PMS1 was cloned. PCR-32 was obtained. The nucleotide sequence can be determined by Taq Dye Primer Cycle Sequenc
ing kit (Applied Biosystems), 373A D
NA Sequencer (Applied Biosystems) was used.

(Example 2) Preparation of human fetal brain cDNA library cDNA synthesized based on human fetal brain mRNA (Clontech) and cloned in a certain direction using UniZAPxR vector kit (Stratagene) A cDNA library with the insert was prepared.

Example 3 Selection of Clone 1 × 10 ^{6 of the} cDNA library prepared in Example 2
Regarding the clone, the clone No. prepared in Example 1 was used.
A hybridization assay was carried out with a probe labeled with ³² P on the PCR-32 DNA fragment. The filter hybridized at 65 ° C overnight and washed at 2XSS
C, 65 ° C. for 15 minutes, autoradiograph was performed under the condition of −80 ° C. overnight to obtain 9 positive clones. The insert DNA of each clone was cloned into pBluescript II SK (-) vector (Stratagene
Company).

(Example 4) Determination of cDNA sequence The nucleotide sequences of the 9 clones obtained in Example 3 were determined using dideoxy.
Method (Hattori M et al., Anal. Biochem. 152, 232-238,
1986) (Sanger F. et al., Proc. Natl. Acad. Sci. U
SA 74, 5463-5467, 1977). As a result, 3 out of 9 clones had the same sequence as the sequence contained in any of the other 6 clones, and as a result, SEQ ID NOs: 1, 2, 3,
Six new sequence cDNAs shown in 4, 5 and 6 were obtained.

(Example 5) Characteristics of cDNA sequence The 6 cDNA sequences obtained in Example 3 are the same in the DNA sequence and the amino acid sequence of the protein encoded by them.
Human PMS2 (Database Accession No.U1
3696, 2771 bp, 862 amino acids). The homology of each DNA sequence and the amino acid sequence of the protein encoded by it with human PMS2 was as follows. SEQ ID NO: 1 (PMS3); 846 bp, 256 amino acids; 95.1
% (Range of 570 bp), 93.3% (range of 179 amino acids) SEQ ID NO: 2 (PMS4); 1151 bp, 252 amino acids; 95.1
% (Range of 570 bp), 92.7% (range of 179 amino acids) SEQ ID NO: 3 (PMS5); 649 bp, 186 amino acids; 92.9
% (Range of 533 bp), 91.5% (range of 176 amino acids) SEQ ID NO: 4 (PMS6); 1007 bp, 159 amino acids; 77.3
% (In the range of 556 bp), 96.2% (in the range of 78 amino acids) SEQ ID NO: 5 (PMS7); 1078 bp, 161 amino acids; 74.5
% (Range of 514 bp), 90.5% (range of 74 amino acids) SEQ ID NO: 6 (PMS8); 1510 bp, 98 amino acids; 87.1
% (Range of 295 bp) and 94.8% (range of 77 amino acids) show high homology, but have clearly different portions, and novel human DN that has not been reported so far.
It was confirmed to be a gene sequence encoding an A mismatch repair-related protein.

(Example 6) Chromosome mapping of genes Cosmid library was screened by using the mixture of cDNAs obtained in Example 4 as a probe and 4 × 10 ⁵
Twenty-six positive clones were obtained from a colony of cosmid clones (corresponding to 5.3 haploid genome). As a result of examining the partial sequences of the bases of these cosmid clones, clones corresponding to SEQ ID NOS: 1, 2, 4 and 5 and clones not corresponding to any of them were included. 0.5 μg each of the DNA of each cosmid was biotin-16-
Labeled by nick translation method using dUTP to be a probe, and chromosome mapping by FISH method
(Takahashi E. et al., Hum. Genet. 86, 14-16, 199
0) went. As a result, 7 in all clones
Signals were observed at q11.23 and 7q22, and both were confirmed to be mapped to the long arm of chromosome 7.

[0024]

[Sequence list]

 SEQ ID NO: 1 Sequence length: 846 Sequence type: Nucleic acid Number of strands: Double strand Topology: Linear Sequence type: cDNAtomRNA Origin Biological name: Homo sapiens Direct origin Library name: Human fetal brain cDNA live Larry sequence characteristics Characteristic symbol: CDS Location: 3. . 773 Method of Characterizing: E Sequence GC CGT GGA GGG AAC TTT CCC AGT CCC CGA GGC GGA TCC GGT GTT GCA 47 Arg Gly Gly Asn Phe Pro Ser Pro Arg Gly Gly Ser Gly Val Ala 1 5 10 15 TCC TTG GAG CGA GCT GAG AGC TCG AGT ACA GAA CCT GCT AAG GCC ATC 95 Ser Leu Glu Arg Ala Glu Ser Ser Ser Thr Glu Pro Ala Lys Ala Ile 20 25 30 AAA CCT ATT GAT CGG AAG TCA GTC CAT CAG ATT TGC TCT GGG CCG GTG 143 Lys Pro Ile Asp Arg Lys Ser Val His Gln Ile Cys Ser Gly Pro Val 35 40 45 GTA CCG AGT CTA AGC ACT GCG GTG AAG GAG TTA GTA GAA AAC AGT CTG 191 Val Pro Ser Leu Ser Thr Ala Val Lys Glu Leu Val Glu Asn Ser Leu 50 55 60 GAT GCT GGT GCC ACT AAT ATT GAT CTA AAG CTT AAG GAC TAT GGA GTG 239 Asp Ala Gly Ala Thr Asn Ile Asp Leu Lys Leu Lys Asp Tyr Gly Val 65 70 75 GAT CTC ATT GAA GTT TCA GGC AAT GGA TGT GGG GTA GAA GAA GAA AAC 287 Asp Leu Ile Glu Val Ser Gly Asn Gly Cys Gly Val Glu Glu Glu Asn 80 85 90 95 TTC GAA GGC TTA ACT CTG AAA CAT CAC ACA TCT AAG ATT CAA GAG TTT 335 Phe Glu Gly Leu Thr Leu Lys His His Th r Ser Lys Ile Gln Glu Phe 100 105 110 GCC GAC CTA CCT CAG GTT GAA ACT TTT GGC TTT CGG GGG GAA GCT CTG 383 Ala Asp Leu Pro Gln Val Glu Thr Phe Gly Phe Arg Gly Glu Ala Leu 115 120 125 AGC TCA CTT TGT GCA CTG AGT GAT GTC ACC ATT TCT ACC TGC CAT GTA 431 Ser Ser Leu Cys Ala Leu Ser Asp Val Thr Ile Ser Thr Cys His Val 130 135 140 TCG GCG AAG GTT GGG ACT CGA CTG GTG TTT GAT CAC TAT GGG AAA ATC 479 Ser Ala Lys Val Gly Thr Arg Leu Val Phe Asp His Tyr Gly Lys Ile 145 150 155 ATC CAG AAA ACC CCC TAC CCC CAC CCC AGA GGG ATG ACA GTC AGT GTG 527 Ile Gln Lys Thr Pro Tyr Pro His Pro Arg Gly Met Thr Val Ser Val 160 165 170 175 AAG CAG TTA TTT TCT ACG CTA CCT GTG CAC CAT AAA GAA TTT CAA AGG 575 Lys Gln Leu Phe Ser Thr Leu Pro Val His His Lys Glu Phe Gln Arg 180 185 190 AAT ATT AAG AAG AAA CGT GCC TGC TTC CCC TTC GCC TTC TGC CGT GAT 623 Asn Ile Lys Lys Lys Arg Ala Cys Phe Pro Phe Ala Phe Cys Arg Asp 195 200 205 TGT CAG TTT CCT GAG GCC TCC CCA GCC ATG CTT CCT GTA CAG CCT GCA 671 Cys Gln Phe Pro Glu Ala Se r Pro Ala Met Leu Pro Val Gln Pro Ala 210 215 220 GAA CTG ACT CCT AGA AGT ACC CCA CCC CAC CCC TGC TCC TTG GAG GAC 719 Glu Leu Thr Pro Arg Ser Thr Pro Pro His Pro Cys Ser Leu Glu Asp 225 230 235 AAC GTG ATC ACT GTA TTC AGC TCT GTC AAG AAT GGT CCA GGT TCT TCT 767 Asn Val Ile Thr Val Phe Ser Ser Val Lys Asn Gly Pro Gly Ser Ser 240 245 250 255 AGA TGA TCTGCAAAAA TGGCTCCTCT CCTCCTTCCT TATGTCTCCA TTAGCACTGG 823GTCCT 256TA

SEQ ID NO: 2 Sequence length: 1151 Sequence type: Nucleic acid Number of strands: Double strand Topology: Linear Sequence type: cDNAtomRNA Origin organism name: Homo sapiens Direct origin Library name: Human fetus Brain cDNA library Sequence features Characteristic symbols: CDS Location: 2. . 760 Method of characterizing: E sequence C TTT TCC AGT TCC CGA GGC GGA TCC GGT GTT GCA TCC TTG GAG CGA 46 Phe Ser Ser Ser Arg Gly Gly Ser Gly Val Ala Ser Leu Glu Arg 1 5 10 15 GCT GAG AGC TGG AGT ACA GAA CCT GCT AAG GCC ATC AAA CCT ATT GAT 94 Ala Glu Ser Trp Ser Thr Glu Pro Ala Lys Ala Ile Lys Pro Ile Asp 20 25 30 CGG AAG TCA GTC CAT CAG ATT TGC TCT GGG CCG GTA GTA CTG AGT CTA 142 Arg Lys Ser Val His Gln Ile Cys Ser Gly Pro Val Val Leu Ser Leu 35 40 45 AGC ACT GCG GTG AAG AAG ATT GTA GAA AAC AGT CTG GAT GCT GGT GCC 190 Ser Thr Ala Val Lys Lys Ile Val Glu Asn Ser Leu Asp Ala Gly Ala 50 55 60 ACT AAT ATT GAT CTA AAG CTT AAG GAC TAT GGA GTG GAT CTC ATT GAA 238 Thr Asn Ile Asp Leu Lys Leu Lys Asp Tyr Gly Val Asp Leu Ile Glu 65 70 75 GTT TCA GGC AAT GGA TGT GGG GTA GAA GAA GAA AAC TTC GAA GGC TTA 286 Val Ser Gly Asn Gly Cys Gly Val Glu Glu Glu Asn Phe Glu Gly Leu 80 85 90 95 ACT CTG AAA CAT CAC ACA TCT AAG ATT CAA GAG TTT GCC GAC CTA CCT 334 Thr Leu Lys His His Thr Ser Lys Ile Gln Glu Phe Ala Asp Leu Pro 100 105 110 CAG GTT GAA ACT TTT GGC TTT CGG GGG GAA GCT CTG AGC TCA CTT TGT 382 Gln Val Glu Thr Phe Gly Phe Arg Gly Glu Ala Leu Ser Ser Leu Cys 115 120 125 GCA CTG AGT GAT GTC ACC ATT TCT ACC TGC CAC GTA TCG GCG AAG GTT 430 Ala Leu Ser Asp Val Thr Ile Ser Thr Cys His Val Ser Ala Lys Val 130 135 140 GGG ACT CGA CTG GTG TTT GAT CAC GAT GGG AAA ATC ATC CAG AAA ACC 478 Gly Thr Arg Leu Val Phe Asp His Asp Gly Lys Ile Ile Gln Lys Thr 145 150 155 CCC TAC CCC CAC CCC AGA GGG ACC ACA GTC AGC GTG AAG CAG TTA TTT 526 Pro Tyr Pro His Pro Arg Gly Thr Thr Val Ser Val Lys Gln Leu Phe 160 165 170 175 TCT ACG CTA CCT GTG CAC CAT AAA GAA TTT CAA AGG AAT ATT AAG AAG 574 Ser Thr Leu Pro Val His His Lys Glu Phe Gln Arg Asn Ile Lys Lys 180 185 190 AAA CGT GCC TGC TTC CCC TTC GCC TTC TGC CGT GAT TGT CAG TTC CTT 622 Lys Arg Ala Cys Phe Pro Phe Ala Phe Cys Arg Asp Cys Gln Phe Leu 195 200 205 GAG GGC TCC CCA GCC ATG CTT CCT GTA CAG CCT GCA AAA CTG ACT CCT 670 Glu Gly Ser Pro Ala Met Le u Pro Val Gln Pro Ala Lys Leu Thr Pro 210 215 220 AGA AGT ACC CCA CCC CAC CCC TGC TCC TTG GAG GAC AAC GTG ATC ACT 718 Arg Ser Thr Pro Pro His Pro Cys Ser Leu Glu Asp Asn Val Ile Thr 225 230 235 GTA TTC AGC TCT GTC AAG AAT GGT CCA GGT TCT TCT AGA TGA 760 Val Phe Ser Ser Val Lys Asn Gly Pro Gly Ser Ser Arg 240 245 250 252 TCTGCAAGAGTCAT GATCACAGTC ATTAGCATTG GAATAAAGTT 820 CCTGCTGATCATCTCGAGAA ATCCACATCT CCCC TCAAGTCTAG 940 CTAGAGTTCA GAGCAATTGA GAAGTGCAAT TTTATCTCCT GCCTTTCATT CTATACCCTG 1000 CTTCTGAACC ATCGTGTTCA ACTGTGAAAC TCACACTTTG GTGACCCTGA CTCCAAAACT 1060 TAATACACCC AAGGTCAGCC CCAGTGATCT GCTTCGA120TCAGATCAG

SEQ ID NO: 3 Sequence length: 649 Sequence type: Nucleic acid Number of strands: Double strand Topology: Linear Sequence type: cDNAtomRNA Origin organism name: Homo sapiens Direct origin Library name: Human fetus Brain cDNA library Sequence features Characteristic symbols: CDS Location: 1. . 561 Method of characterizing: E sequence TCC AGT ACA GAA CCT GCT AAG GCC ATC AAA CCT ATT GAT CGG AAG TCA 48 Ser Ser Thr Glu Pro Ala Lys Ala Ile Lys Pro Ile Asp Arg Lys Ser 1 5 10 15 GTC CAT CAG ATT TGC TCT GGG CCA GTG GTA CTG AGT CTA AGC ACT GCA 96 Val His Gln Ile Cys Ser Gly Pro Val Val Leu Ser Leu Ser Thr Ala 20 25 30 GTG AAG GAG TTA GTA GAA AAC AGT CTG GAT GCT GGT GCC ACT AAT ATT 144 Val Lys Glu Leu Val Glu Asn Ser Leu Asp Ala Gly Ala Thr Asn Ile 35 40 45 GAT CTA AAG CTT AAG GAC TAT GGA GTG GAT CTC ATT GAA GTT TCA GAC 192 Asp Leu Lys Leu Lys Asp Tyr Gly Val Asp Leu Ile Glu Val Ser Asp 50 55 60 AAT GGA TGT GGG GTA GAA GAA GAA AAC TTT GAA GGC TTA ACT CTT TCA 240 Asn Gly Cys Gly Val Glu Glu Glu Asn Phe Glu Gly Leu Thr Leu Ser 65 70 75 80 GCT CTG AAA CAT CAC ACA TGT AAG ATT CAA GAG TTT GCC GAC CTA ACT 288 Ala Leu Lys His His Thr Cys Lys Ile Gln Glu Phe Ala Asp Leu Thr 85 90 95 GAA GTT GAA ACT TTC GGT TTT CAG GGG GAA GCT CTG AGC TCA CTG TGT 336 Glu Val Glu Thr Phe Gly Phe Gln G ly Glu Ala Leu Ser Ser Leu Cys 100 105 110 GCA CTG AGC GAT GTC ACC ATT TCT ACC TGC CAC GCG TCG GTG AAG GTT 384 Ala Leu Ser Asp Val Thr Ile Ser Thr Cys His Ala Ser Val Lys Val 115 120 125 GGG ACT CGA CTG GTG TTT GAT CAC GAT GGG AAA ATC ATC CAG GAA ACC 432 Gly Thr Arg Leu Val Phe Asp His Asp Gly Lys Ile Ile Gln Glu Thr 130 135 140 CCC TAC CCC CCA CCC CAG AGG ACC ACA GTC AGC GTG AAG CAG TTA TTT 480 Pro Tyr Pro Pro Pro Gln Arg Thr Thr Val Ser Val Lys Gln Leu Phe 145 150 155 160 TCT ACG CTA CCT GTG CGC CAT AAG GAA TTT CAA AGG AAT ATT AAG AAG 528 Ser Thr Leu Pro Val Arg His Lys Glu Phe Gln Arg Asn Ile Lys Lys 165 170 175 ACG TGC CTG CTT CCC CTT CGC CTT CTG CCG TGA TTGTCAGTTT CCTGAGGCCT 581 Thr Cys Leu Leu Pro Leu Arg Leu Leu Pro 180 185 186 CCCCAGCCAT GCTTCCTGTA CAGCCTGCAG AACTGTGAGC CAATTAAACC TCATATTCTCTCT

SEQ ID NO: 4 Sequence length: 1007 Sequence type: Nucleic acid Number of strands: Double strand Topology: Linear Sequence type: cDNAtomRNA Origin organism name: Homo sapiens Direct origin Library name: Human fetus Brain cDNA library Sequence features Characteristic symbols: CDS Location: 1. . 480 Method of characterizing: E sequence CGG GTA TTG AGT CCA TGG AGC GAG CTG AGA GCT AGA GGT ACA GAA CCT 48 Arg Val Leu Ser Pro Trp Ser Glu Leu Arg Ala Arg Gly Thr Glu Pro 1 5 10 15 GCT AAG GCC ATC AAA CCT ATC GAT CGG AAG TCA GTC CAT CAG ATT TGC 96 Ala Lys Ala Ile Lys Pro Ile Asp Arg Lys Ser Val His Gln Ile Cys 20 25 30 TCT GGG CCG GTG GTA CTG AGT CTA AGC ACT GCG GTG AAG GAG TTA GTA 144 Ser Gly Pro Val Val Leu Ser Leu Ser Thr Ala Val Lys Glu Leu Val 35 40 45 GAA AAC AGT CTG GAT GCT GGT GCC ACC AAT ATT GAT CTA AAG CTT AAG 192 Glu Asn Ser Leu Asp Ala Gly Ala Thr Asn Ile Asp Leu Lys Leu Lys 50 55 60 GAC TAT GGA GTG GAT CTC ATT GAA GTT TCA GAC AAT GGA TGT GGG GTA 240 Asp Tyr Gly Val Asp Leu Ile Glu Val Ser Asp Asn Gly Cys Gly Val 65 70 75 80 GAA GAA GAA AAC TTT GAA GGC TTA ACG ATG TCA CCA TTT CTA CCT GCC 288 Glu Glu Glu Asn Phe Glu Gly Leu Thr Met Ser Pro Phe Leu Pro Ala 85 90 95 ACG CAT CGG CGA AGG TTG GGA CTC GAC TGG TGT TTG ATC ACG ATG GGA 336 Thr His Arg Arg Arg Leu Gly Leu A sp Trp Cys Leu Ile Thr Met Gly 100 105 110 AAA TCA TCC AGA AAA CCC CCT ACC CCC ACC CCA GAG GAC CAC AGT CAG 384 Lys Ser Ser Arg Lys Pro Pro Thr Pro Thr Pro Glu Asp His Ser Gln 115 120 125 CGT GAA GCA GTT ATT TCT ACA CTA CCT GTG CAC CAT AAG GAA TTT CAA 432 Arg Glu Ala Val Ile Ser Thr Leu Pro Val His His Lys Glu Phe Gln 130 135 140 AGG AAT ATT AAG AAG ACG TGC CTG TTT CCC CTT CGC CTT CTG CCG TGA 480 Arg Asn Ile Lys Lys Thr Cys Leu Phe Pro Leu Arg Leu Leu Pro 145 150 155 159 TTGTCAGTTT CCTGAGGCCT CCCCAGCCAT GCTTCCTGTA CAGCCTGCAG AACTGGACCA 540 TTGCAAGCTG CAGAAAATGA GAAGACCCTA CTGACCAGCC CAAGAAGACA GATGACAAGA 600 TACTGAGGAT GGCCGTATAA GAAAATAAAC AACTTGCTAA AACTGCTGAA ACTCCCTCTG 660 CTTGTGAGAT TTAAAAAAAC TGGCTGAAAT TGGTTGGAAC CAACATGTCC AATGGAGTCT 720 GAATAGAAAG AGCTTGCTGA CATCACAGCC CGAATTTCCA CCACATTTCA TACTAACTCC 780 CCCTGAATTT GCATATGCAA ACCACAAGCA AGCATGATCA AACAGACAAC TGCGCGTGTC 840 CGAGGACTTT CCAGATCTCC CCTTTCCTTC CACCAATCAC TTACTAATCT GAGAATCCAT 900 CCCCTAATAA AATCACTGCC TTAAAG ACAGGAAGA CAAATTTGAG CTGGACTCAG 960 TGTCTCTTTA TTAGTCGACT TGCAATAAAA AAGCTTTCTT TTCCCCC 1007

SEQ ID NO: 5 Sequence length: 1078 Sequence type: Nucleic acid Number of strands: Double strand Topology: Linear Sequence type: cDNAtomRNA Origin organism name: Homo sapiens Direct origin Library name: Human fetus Brain cDNA library Sequence features Characteristic symbols: CDS Location: 2. . 487 Method of characterizing: E sequence T GCT AAG GCC ATC AAA CCT ATT GAT CGG AAG TCA GTC CAT CAG ATT TGC 49 Ala Lys Ala Ile Lys Pro Ile Asp Arg Lys Ser Val His Gln Ile Cys 1 5 10 15 TCT GGG CCG GTA GTA CTG AGT CTA AGC ACT GCG GTG AAG AAG ATG GTA 97 Ser Gly Pro Val Val Leu Ser Leu Ser Thr Ala Val Lys Lys Met Val 20 25 30 GAA AAC AGT CTG GAT GCT GGT GCC ACT AAT ATT GAT CTA AAG CTT AAG 145 Glu Asn Ser Leu Asp Ala Gly Ala Thr Asn Ile Asp Leu Lys Leu Lys 35 40 45 GAC TAT GGA ATG GAT CTC ATT GAA GTT TCA GGC AAT GGA TGT GGG GTA 193 Asp Tyr Gly Met Asp Leu Ile Glu Val Ser Gly Asn Gly Cys Gly Val 50 55 60 GAA GAA GAA AAC TTC GAA GGC TTA ATG ATG TCA CCA TTT CTA CCT GCC 241 Glu Glu Glu Asn Phe Glu Gly Leu Met Met Ser Pro Phe Leu Pro Ala 65 70 75 80 ACG TAT CGG CGA AGG TTG GGA CTC GAC TGG TGT TTG ATC ACG ATG GGA 289 Thr Tyr Arg Arg Arg Leu Gly Leu Asp Trp Cys Leu Ile Thr Met Gly 85 90 95 AAA TCA TCC AGA AAA CCC CCT ACC CCC ACC CCA GAG GAC CAC AGT CAG 337 Lys Ser Ser Arg Lys Pro Pro Thr Pro Thr Pro Glu Asp His Ser Gln 100 105 110 CGT GAA GCA GTT ATT TCT ACG TAC TGT GCG CAT AAG GAA TTT CAA AGG 385 Arg Glu Ala Val Ile Ser Thr Tyr Cys Ala His Lys Glu Phe Gln Arg 115 120 125 AAT ATT AAG AAG AAA CGT GCC TGC TTC CCC TTC GCC TTC TGC CGT ATT 433 Asn Ile Lys Lys Lys Arg Ala Cys Phe Pro Phe Ala Phe Cys Arg Ile 130 135 140 GTC AGT TTC TTG AGG GCT CCC CAG CCA TGC TTC CTG TAC AGC CTG CAA 481 val Ser Phe Leu Arg Ala Pro Gln Pro Cys Phe Leu Tyr Ser Leu Gln 145 150 155 160 AAC TGA CTCCTAGAAG TACCCCACCC CACCCCTGCT CCTTGGAGGA CAACGTGATC 537 Asn 161 ACTGTATTCA GCTCTGTCAA GAATGGTCCA GGTTCCTCTC CTCCTTCCTG ATGTCTGCCA 597 TTAGCATTGG AATAAAGTTC CTGCTGAAAA TCCACATCTC CCCTGGGTCC GGTGTCTGGA 657 AGTGAGAGAG ACAATGTCAC ACTTCAAGGA GGCAGCTCTC TAGACAGGAA GGTTATTCAC 717 GTCCCATGTC AAGTCTAGCT AGAGTTCAGA GCAATTGAGA AGTGCAATTT TATCTCCTGC 777 CTTTCATTCT ATACCCTGCT TCTGAACCAT CGTGTTCAAC TGTGAAACTC ACACTTTGGT 837 GACCCTGACT CCAAAACTTA ATACACCCAA GGTCAGCCCC AGTGAATCGGTCTCATAC AGGGAGTAG GGCACCCTCA GAGAATGTGG CTTTGGACTT 957 CATCACAGCT GGGGCCTTTT GTGTCACTTC AGATCTAAAC TTGTAACCGT GCTAGATCTG 1017 TTTCTAACGT GACAACATCA CGAACCACGA GTCCAGAAGC CTAATCCATA ATCCTCCCCCCC 1077 A 1078

SEQ ID NO: 6 Sequence length: 1510 Sequence type: Nucleic acid Number of strands: Double strand Topology: Linear Sequence type: cDNAtomRNA Origin organism name: Homo sapiens Direct origin Library name: Human fetus Brain cDNA library Sequence features Characteristic symbols: CDS Location: 1. . 297 Method of characterizing: E sequence ATA CTT GCT TTG TTT CTT GTA ACT GAT TTC TTT AGT ACA GAA CCT GCT 48 Ile Leu Ala Leu Phe Leu Val Thr Asp Phe Phe Ser Thr Glu Pro Ala 1 5 10 15 AAG GCC ATC AAA CCT ATT GAT CGG AAG TCA GTC CAT CAG ATT TGC TCT 96 Lys Ala Ile Lys Pro Ile Asp Arg Lys Ser Val His Gln Ile Cys Ser 20 25 30 GGG CCG GTG GTA CCG AGT CTA AGC ACT GCG GTG AAG GAG TTA GTA GAA 144 Gly Pro Val Val Pro Ser Leu Ser Thr Ala Val Lys Glu Leu Val Glu 35 40 45 AAC AGT CTG GAT GCT GGT GCC ACT AAT ATT GAT CTA AAG CTA AAG GAC 192 Asn Ser Leu Asp Ala Gly Ala Thr Asn Ile Asp Leu Lys Leu Lys Asp 50 55 60 TAT GGA GTG GAT CTC ATT GAA GTT TCA GGC AAT GGA TGT GGG GTA GAA 240 Tyr Gly Val Asp Leu Ile Glu Val Ser Gly Asn Gly Cys Gly Val Glu 65 70 75 80 GAA GAA AAC TTC GAA GGC TTA AGT AAG TTA ACT TTT TCT AAT CCT ATT 288 Glu Glu Asn Phe Glu Gly Leu Ser Lys Leu Thr Phe Ser Asn Pro Ile 85 90 95 ATA AAA TAA TTGGGCCACA TGTCTTAGAA TTTTGAGTAA CACTGTCTTG GGAAACACAA 347 Ile Lys 98 AAACAGTTTT TTAAAGCC AG TTACTAGATA TCATGTATAT TTGTTGTTAT AGCACTTAAG 407 ATATCTTAGT CCTTACTTTA TACTCTCTTT CAGCTCTGAA ACATCACACA TCTAAGATTC 467 AAGAGTTTGC CGACCTACCT CAGGTTGAAA CTTTTGGCTT TCGGGGGGAA GCTCTGAGCT 527 CACTTTGTGC ACTGAGGTGA TAAAATATTT TTATCCATTC ACTTGACCCT TAGAAAAACC 587 TCTCTGAAAA TTAATTGGAA TCATTATTAT TTACAGTTTT CTGTCTCAAT ATCTCAGCTT 647 CCAGCTTCTT GAATTCTGTT TTGTCTCACT GCCAATCTAA GTCCTAGTAC TTCTGAAATG 707 TGAGCAATAA ATGAATGAAA TGAAGCAAAT AGTATTGTTA AAAAAATTGG TTACCCTTAT 767 TAAAACAGTA ACTTCTCAAT TTGAACATAA CATATAGATA ATAAATGATA GTTACCATTG 827 GTTTTCATTA TCAATTTTTA GGGAAACATT TCACCAAAGC ACTATTTAAT TATAGCACAG 887 ATACTAAATT TTTATAAATA ATTATATGCA CACACATACA CAGCTAAATT AGAAAACACA 947 ATTTTAAATT TTATATAATT ACTAGTTATT AATGGGTAAA ATTCATTTTT CTTTTAATAT 1007 CTAAAATATT ATTTATGCCC CGAAATATGC TAATAAAAGT TAAAGATTGT GTATTATGCC 1067 TAGATTCAAA GTTTTTGGCA AACTTCAAAT ACTTTTGATT GACATGTACA TTAGCAGTTA 1127 TTAGTTGACT TTTACTAAAA CATGCACCTT TACGATGGTG TTTTTGTAGA AACATAAGTA 1187 GTCAAATAGT AACATTTATT GCAGTTTTGT A CAGTATTTG AGTATAGTGC ATAAATAGGT 1247 ATGTTCATAA AATCAATAAA AGGAGTATCT ATGGAAATAA GCAAAATATC AGCAAAAATT 1307 GCTATTCCTT TGCCTCATTT AATTTTTAAA TTCTTTGTTC CATGTGATTC TATTATATGC 1367 CTTTGGCATG CCTTGTATAA AACCAAGGTC ACAAATGCAA CTTATTAACG GAATTTGCCA 1427 AGAAGACAGC CTTGACATTA AAAATTCCCT CACAAGAGGT AATCCATAAT CCTTAATAAA 1487 ATAAAATTGT TACATGAGCC AAT 1510

[Brief description of drawings]

FIG. 1 is a phylogenetic tree resulting from sequence comparison of major known DNA mismatch repair-related proteins.

FIG. 2: Known DNA mismatch repair-related protein, yeast P
A phylogenetic tree based on the result of sequence comparison of MS1, human PMS1, human PMS2 and protein PMS3, 4, 5, 6, 7, 8 encoded by this DNA.

Continuation of the front page (51) Int.Cl. ⁶ Identification number Office reference number FI Technical display location C12N 5/10 15/09 ZNA C12P 21/02 C 9282-4B

Claims (10)

[Claims] 1. A DNA represented by SEQ ID NO: 1, 2, 3, 4, 5 or 6, or a DN comprising the DNA.
A 2. A peptide encoded by the DNA shown in SEQ ID NO: 1, 2, 3, 4, 5 or 6, or a peptide comprising the peptide. 3. A DNA comprising a part of the DNA shown in SEQ ID NO: 1, 2, 3, 4, 5 or 6. 4. A peptide comprising a part of the peptide encoded by the DNA shown in SEQ ID NO: 1, 2, 3, 4, 5 or 6. 5. A plasmid containing the DNA according to claim 1 or 3. 6. A transformant carrying the plasmid according to claim 5. 7. The method for producing a peptide according to claim 2 or 4, which comprises culturing the transformant according to claim 6 and recovering the expression product. 8. An antibody which binds to the peptide according to claim 2 or 4. 9. A primer or probe containing a part of the DNA sequence according to claim 1 or 3. 10. A method for gene analysis, which comprises hybridizing the primer or probe according to claim 9 with a test DNA.