KR101071215B1 - Use of snp for diagnosing type ii diabetes mellitus - Google Patents

Abstract

The present invention provides the use of SNPs for diagnosing type 2 diabetes. More specifically, the present invention relates to a type 2 diabetes diagnostic polynucleotide comprising the SNP of the PSMB5 gene or a complementary polynucleotide thereof, a polynucleotide hybridizing thereto, a polypeptide encoded by the same, a microarray and a kit comprising the polynucleotide, Provided are a method for diagnosing type 2 diabetes, a method for screening a medicine for treating type 2 diabetes, and a pharmaceutical composition for treating type 2 diabetes.

The rs2230087 polymorphic genotype of the PSMB5 gene of the present invention can be usefully used as a marker for diagnosing the genetic sensitivity of diabetes mellitus specifically.

PSMB5, SNP, Type 2 Diabetes

Description

Use of SNP for diagnosis of type 2 diabetes {USE OF SNP FOR DIAGNOSING TYPE II DIABETES MELLITUS}

The present invention provides the use of SNPs for diagnosing type 2 diabetes. More specifically, the present invention relates to a type 2 diabetes diagnostic polynucleotide comprising the SNP of the PSMB5 gene or a complementary polynucleotide thereof, a polynucleotide hybridizing thereto, a polypeptide encoded by the same, a microarray and a kit comprising the polynucleotide, Provided are a method for diagnosing type 2 diabetes, a method for screening a medicine for treating type 2 diabetes, and a pharmaceutical composition for treating type 2 diabetes.

The 26S ubiquitin-proteasome system (UPS) is a major proteolytic pathway in eukaryotic cells that selectively mediates proteolysis in relation to cell death, cell cycle, cell differentiation and proliferation and inflammation. It is known to regulate physiological functions (Pickart, CM (2004) Back to the future with ubiquitin Cell. 116,181-1902). Proteolysis by UPS acts as a "quality-control" that quickly removes proteins that harm cell function and survival, such as non-folding and damaged proteins. Neuropathy, such as Alzheimer's disease or Parkinson's disease, and hereditary neurological diseases such as Huntington's disease have been reported (Jiang, Y and Beaudet, AL (2004) Human disorders of ubiquitination and proteasomal degradation.Cur Opin Pediatr. 16, 419-426).

UPS undergoes a ubiquitination process in which the substrate protein to be degraded through a series of reactions of three enzymes is labeled with ubiquitin. and Ciechnover, A. (1998) The Ubiquitin System.Annu. Rev. Biochem. 67, 425-479), belonging to IkB and Insulin receptor substrate (IRS). IkB complexes with NF-kB in the cytoplasm and inhibits NF-kB activity, and it is believed that IkB is degraded by UPS to increase NFkB activity and affect insulin resistance (Yamamoto Y, Gaynor). RB: IkappaB kinases: key regulators of the NFkappaB pathway.Trends Biochem Sci 29: 7279, 2004; Yuan M, Konstantopoulos N, Lee J, Hansen L, Li ZW, Karin M, Shoelson SE: Reversal of obesity- and diet-induced insulin resistance with salicylates or targeted disruption of Ikkbeta Science 293:. 16731677, 2001). IRS is also a protein that plays an important role in the insulin signaling process and is known to affect insulin resistance by the degradation of IRS protein by 26S after ubiquitination (White, MF 1997. The insulin signaling system and the IRS proteins.Diabetologia 40 (Suppl. 2): S2S17; White, MF 1998.The IRS-signaling system: a network of docking proteins thatmediate insulin and cytokine action.Recent Prog.Hormone Res. 53: 119-138; Curr Opin Clin Nutr Metab Care. 2004 May; 7 (3): 249-54.The ubiquitin-proteasome pathway is a new partner for the control of insulin signaling.Sophie Rome (Rome S.), Emmanuelle Meugnier and Hubert Vidal.

The 26S proteasome is a large multicatalytic protease of about 2.5 MDa that is present in the cytoplasm and nucleus and consists of a catalytically active 20S subunit and two 19S regulators (Jiang, Y and Beaudet, AL). (2004) Human disorders of ubiquitination and proteasomal degradation.Cur Opin Pediatr. 16, 419-426). 20S consists of seven alpha subunits that play a structural role and seven beta subunits that play a key role in catalytic activity.

The beta subunit has the activity of three proteases: chymotrypsin-like protease, trypsin-like protease, and post-glutamyl protease. Of these proteases, chymotrypsin-like proteases are proteolytic rate modulators, and inhibition of chymotrypsin-like proteases can inhibit almost all activities of proteasomes (Molecular Cell, Vol. 4, 395402, September, 1999). It is the activity of proteasome subunit β type 5 (PSMB 5) in the beta subunit, and Bortezomib, which reversibly binds to PSMB5 and inhibits the activity of chymotrypsin-like protease, inhibits the degradation of I-kB which is bound to NF-kB. Has been shown to reduce NF-kB activity (Lightcap ES, McCormack TA, Pien CS, Chau V, Adams J, Elliott PJ.Proteasome inhibition measurements: clinical application.Clin Chem 2000; 46: 673-83; Hideshima T , Mitsiades C, Akiyama M, Hayashi T, Chauhan D, Richardson P, et al. Molecular mechanisms mediating antimyeloma activity of proteasome inhibitor PS-341. Blood 2003; 101: 1530-4).

The present inventors thought that the polymorphism of the PSMB 5 gene would affect diabetes through IRS as well as NF-kB. Therefore, we tried to study polymorphism in the PSMB5 gene in patients with type 2 diabetes.

Single nucleotide polymorphism (SNP) is a single base-pair variation in DNA between individuals of the same species, the most common form of gene polymorphism. SNPs present in the transcriptional regulatory base sequences, such as promoters, can regulate the amount of gene expressed. Base sequences in exon-intron boundaries that affect RNA splicing or SNPs in 3'-UTR may affect RNA stability or translation efficiency. When SNPs occur in the coding region, either of the polymorphic forms can express a protein that is defective in function by mutation. If SNPs occur in the non-coding region, they may not affect the phenotype, or may otherwise express defective proteins as a result of defective splicing. When the SNP affects the phenotype and the expression of the disease varies depending on the difference in the phenotype, the SNP may be used as a marker indicating the susceptibility to the disease. As a result of the completion of the human genome map, studies on the presence of SNPs have also been published in databases such as GenBank, but most of the effects of the presence of SNPs on the phenotype have not been studied. The present inventors investigated the correlation between the allele type of SNP (rs2230087) located at 3'-UTR of PSMB5 gene exon3 and type 2 diabetes in order to determine the effect of the polymorphism of PSMB5 gene on the onset of type 2 diabetes. The invention was completed.

An object of the present invention is to provide an SNP that can be used as a marker for diagnosing susceptibility to type 2 diabetes.

Another object of the present invention is to provide a polynucleotide comprising the SNP or a complementary polynucleotide thereof.

Another object of the present invention is to provide a polynucleotide that specifically hybridizes with the polynucleotide comprising the SNP or its complementary polynucleotide.

Another object of the present invention is to provide a polypeptide encoded by the polynucleotide comprising the SNP or a complementary polynucleotide thereof.

Another object of the present invention is a type 2 diabetes comprising a polynucleotide comprising the SNP or a complementary polynucleotide thereof, a polynucleotide specifically hybridizing thereto, or a polypeptide specifically encoded by the same or a cDNA thereof. It is to provide a composition for diagnosing diseases.

Another object of the present invention is for diagnosing type 2 diabetes comprising the polynucleotide comprising the SNP or the complementary polynucleotide thereof, the polynucleotide specifically hybridizing therewith, the polypeptide specifically encoded therein or the cDNA thereof. It is to provide a microarray.

Still another object of the present invention is to provide a kit for diagnosing type 2 diabetes comprising the microarray.

Another object of the present invention is to contact a polynucleotide comprising the SNP or a polypeptide encoded by the complementary polynucleotide thereof with a test substance; And it provides a screening method of a medicament for treating type 2 diabetes comprising the step of determining whether the test substance exhibits the activity to enhance or inhibit the function of the polypeptide.

Still another object of the present invention is to provide a pharmaceutical composition for treating type 2 diabetes, wherein the base of the SNP position of NCBI refSNP ID: rs2230087 includes a PSMB5 gene having a GG genotype.

The present invention provides a polynucleotide for diagnosis of type 2 diabetes or its complementary polynucleotide consisting of 10 or more consecutive bases including the base of the SNP position of NCBI refSNP ID: rs2230087 of the polynucleotide consisting of the nucleotide sequence of SEQ ID NO: 1. to provide.

The NCBI refSNP ID for the SNP indicates the sequence of the SNP and its position. Those skilled in the art can easily identify the position and sequence of the SNP using the number. It will be apparent to those skilled in the art that the specific sequence corresponding to the refSNP ID of the SNP registered in the NCBI may be slightly changed depending on the results of the subsequent gene studies, and such altered sequences are also included within the scope of the present invention.

As can be seen from the following examples, gene polymorphisms in the 3'-untranslated region of exon 3 in the PSMB5 gene were studied in normal and type 2 diabetic patients. There is a SNP of NCBI refSNP ID: rs2230087 associated with the A allele, and the GA genotype or AA genotype at the SNP position is significantly higher in the type 2 diabetic group than the normal control group.

Therefore, the SNP of the NCBI refSNP ID: rs2230087 of the PSMB5 gene can be used as a diagnostic marker for susceptibility to type 2 diabetes.

Accordingly, the present invention provides a polynucleotide for diagnosis of type 2 diabetes or its complementary polynucleotide comprising 10 or more consecutive bases including the base of the SNP position of NCBI refSNP ID: rs2230087 of the polynucleotide consisting of the nucleotide sequence of SEQ ID NO: 1. To provide.

The base sequence of SEQ ID NO: 1 is a polymorphic sequence including a polymorphic site representing a SNP in a polynucleotide sequence. In the present invention, the polynucleotide may be DNA or RNA.

Type 2 diabetes diagnostic polynucleotide or its complementary polynucleotide comprising the SNP according to the present invention is preferably composed of 10 or more consecutive bases.

The present invention also provides a polynucleotide that specifically hybridizes to a type 2 diabetes diagnostic polynucleotide or its complementary polynucleotide comprising the SNP.

In the present invention, the polynucleotide that specifically hybridizes to the type 2 diabetes diagnostic polynucleotide or its complementary polynucleotide is an allele-specific polynucleotide.

Allele specific polynucleotide means specific hybridization to each allele. That is, it means hybridizing to specifically distinguish bases of the polymorphic sites present in the polymorphic sequence. Here, hybridization is usually carried out under stringent conditions, for example salt concentrations of 1 M or less and temperatures of 25 ° C. or higher. For example, conditions of 5 μs SSPE (750 mM NaCl, 50 mM Na Phosphate, 5 mM EDTA, pH 7.4) and 25-30 ° C. may be suitable for allele specific probe hybridization.

In the present invention, the allele specific polynucleotide may be an allele specific probe. In the present invention, the probe means a hybridization probe, and means an oligonucleotide capable of sequence-specific binding to the complementary strand of a nucleic acid. The allele-specific probe of the present invention has a polymorphic site in nucleic acid fragments derived from two individuals of the same species, and hybridizes to a DNA fragment derived from one individual but not to a fragment derived from another individual. In this case, the hybridization conditions should be severe enough to hybridize to only one of the alleles, since the hybridization conditions show a significant difference in hybridization strength between alleles. In the probe of the present invention, the central region is preferably aligned with the polymorphic region of the polymorphic sequence. This can lead to good hybridization differences between different allelic forms. The probe of the present invention can be used in diagnostic kits or diagnostic methods such as microarrays for detecting alleles and diagnosing susceptibility to type 2 diabetes.

In addition, in the present invention, the allele specific polynucleotide may be an allele specific primer. Appropriate lengths of primers may vary depending on the intended use, but generally consist of 15 to 30 bases. The primer sequence need not be completely complementary to the template but must be sufficiently complementary to hybridize with the template. The primer hybridizes to a DNA sequence comprising a polymorphic site to amplify a DNA fragment comprising a polymorphic site. In the present invention, the primer may be a primer having a nucleotide sequence of SEQ ID NO: 2 or SEQ ID NO: 3, for example.

The present invention also relates to a type 2 diabetes diagnostic polynucleotide or a complementary polynucleotide consisting of 10 or more consecutive bases comprising a base at the SNP position of NCBI GenBank accession number rs2230087 of a polynucleotide consisting of the nucleotide sequence of SEQ ID NO: 1 It provides a polypeptide encoded by. The polypeptide may be used in a composition for diagnosing type 2 diabetes, a microarray or a kit, a method for diagnosing type 2 diabetes, or a method for screening a medicine for treating type 2 diabetes. Alternatively, an antibody against the polypeptide may be used in place of the polypeptide. It is preferable that it is a monoclonal antibody as said antibody.

The present invention also relates to a type 2 diabetes diagnostic polynucleotide or a complementary polynucleotide consisting of 10 or more consecutive bases comprising a base at the SNP position of NCBI GenBank accession number rs2230087 of a polynucleotide consisting of the nucleotide sequence of SEQ ID NO: 1 The present invention provides a composition for diagnosing type 2 diabetes, comprising a polynucleotide that hybridizes specifically thereto, or a polypeptide specifically encoded by the polypeptide or cDNA thereof. Examining the genotype of the SNP position of NCBI GenBank Accession No. rs2230087 using the composition enables diagnosis of susceptibility to type 2 diabetes.

Accordingly, the present invention also comprises 10 or more consecutive bases comprising the base of the SNP position of NCBI GenBank Accession No. rs2230087 of a polynucleotide consisting of the base sequence of SEQ ID NO: 1 for the preparation of a microarray or kit for diagnosing type 2 diabetes It provides a use of a composition comprising a polynucleotide for diagnosis of type 2 diabetes or a complementary polynucleotide thereof, a polynucleotide that specifically hybridizes therewith, or a polypeptide or cDNA specifically encoded thereby.

The present invention also provides a method for diagnosing type 2 diabetes using the composition. Diagnosis method of type 2 diabetes in accordance with the present invention comprises the steps of separating the nucleic acid sample from the subject; And determining an allele of the base at the SNP position of NCBI GenBank Accession No. rs2230087 of the polynucleotide consisting of the nucleotide sequence of SEQ ID NO: 1. The nucleic acid sample includes DNA, mRNA, or cDNA synthesized from mRNA.

For example, DNA is isolated from tissues, body fluids, or cells of a subject, and then amplified by PCR, followed by SNP analysis. SNP analysis can be performed by conventional methods known in the art. For example, as in the embodiment of the present invention, the SNP analysis may be performed using a real time PCR system, the direct determination of the nucleotide sequence of the nucleic acid by the dideoxy method, or the sequence of the SNP site may be performed. The nucleotide sequence of the polymorphic site is determined by hybridizing a probe or a complement thereof to the DNA and measuring the degree of hybridization resulting therefrom, or allele-specific probe hybridization, allele specific amplification. Allele-specific amplification, sequencing, 5 'nuclease digestion, molecular beacon assay, oligonucleotide ligation assay, Size analysis and single-stranded conformation polymorphism To be carried out.

The present invention also relates to a type 2 diabetes diagnostic polynucleotide or a complementary polynucleotide consisting of 10 or more consecutive bases comprising a base at the SNP position of NCBI GenBank accession number rs2230087 of a polynucleotide consisting of the nucleotide sequence of SEQ ID NO: 1 A microarray for diagnosing type 2 diabetes, comprising a polynucleotide that specifically hybridizes therewith, or a polypeptide that is specifically encoded therein or a cDNA thereof, is provided. The present invention also includes a type 2 diabetic diagnostic kit comprising the microarray. The microarray and diagnostic kit can be prepared by conventional methods known to those skilled in the art.

In addition, the present invention is a type 2 diabetes diagnostic polynucleotide or complementary polynucleotide consisting of 10 or more consecutive bases comprising the base of the SNP position of NCBI GenBank accession number rs2230087 of the polynucleotide consisting of the nucleotide sequence of SEQ ID NO: 1 10 or more consecutive bases comprising the base of the SNP position of NCBI GenBank Accession No. rs2230087 of a polynucleotide consisting of the composition for screening a medicament for treating type 2 diabetes comprising a polypeptide encoded by Contacting a test object with a polypeptide encoded by a polynucleotide for diagnosis of type 2 or a complementary polynucleotide thereof; And it provides a screening method of a medicament for treating type 2 diabetes comprising the step of determining whether the test substance exhibits the activity to enhance or inhibit the function of the polypeptide.

In the screening method of the present invention, the reaction between the polypeptide and the test substance may be used by conventional methods used to determine whether the reaction between the protein and the protein and the compound is performed. For example, a method of measuring activity after reacting the protein with a test substance, yeast two-hybrid, phage-displayed peptide clone binding to the protein, High throughput screening (HTS) using natural products and chemical libraries, drug hit HTS, cell-based screening, or DNA array (DNA array) Can be used. In this case, the composition of the present invention may include, in addition to the polypeptide, a buffer or a reaction solution which stably maintains the structure or physiological activity of the polypeptide. In addition, the composition of the present invention may include a cell expressing the polypeptide, or a cell containing a plasmid expressing the polypeptide under a promoter capable of controlling transcription rate for in vivo experiments.

In the screening method of the present invention, the test substance may be individual nucleic acids, proteins, other extracts or natural products, compounds, or the like, which are estimated to have the potential as a type 2 diabetes therapeutic agent according to a conventional selection method or are randomly selected. have.

The invention also provides a pharmaceutical composition for treating type 2 diabetes comprising a PSMB5 gene whose base at the SNP position of NCBI GenBank Accession No. rs2230087 has a GG genotype. According to the present invention, if the base at the SNP position of NCBI GenBank Accession No. rs2230087 has a GA or AA genotype, the likelihood of developing type 2 diabetes is high. Therefore, in the case of type 2 diabetes patients with GA or AA genotype, the genotype of SNP of NCBI GenBank Accession No. rs2230087 can be modified to GG genotype to treat type 2 diabetes. Therefore, type 2 diabetes can be treated by introducing a PSMB5 gene whose base at the SNP position of NCBI GenBank Accession No. rs2230087 with GG genotype into cells of type 2 diabetes patients. Introduction of PSMB5 with GG genotype can be carried out by conventional nucleic acid therapies known to those skilled in the art. For example, PSMB5 gene with GG genotype can be introduced into cells via viral or non-viral carriers to modify the genotype of SNP of CBI GenBank Accession No. rs2230087 to GG genotype to treat type 2 diabetes. A therapeutically effective amount of a composition containing the PSMB5 gene of the present invention as an active ingredient means an amount required to achieve the effect of treating type 2 diabetes. Thus, the type of disease, the severity of the disease, the type and amount of the active and other ingredients contained in the composition, the type of formulation and the age, weight, general health, sex and diet, sex and diet, time of administration, route of administration and composition of the patient. It can be adjusted according to various factors including the rate of secretion, the duration of treatment, and the drug used concurrently.

The rs2230087 polymorphic genotype of the PSMB5 gene of the present invention can be usefully used as a marker for diagnosing the genetic sensitivity of diabetes mellitus specifically.

Advantages and features of the present invention and methods for achieving them will be apparent with reference to the embodiments described below in detail. However, the present invention is not limited to the embodiments disclosed below, but may be implemented in various forms. It is provided to fully convey the scope of the invention to those skilled in the art, and the present invention is defined only by the scope of the claims.

EXAMPLES Analysis of PSMB5 Gene Polymorphism in Patients with Type 2 Diabetes by Genetic Polymorphism Reading

(1) Selection of test subject

281 patients with type 2 diabetes who visited Kyungpook National University Hospital were selected as the experimental group and 146 normal patients as the control group. The selection criteria of the normal person was 60 years or older, no history of diabetes mellitus and hypertension, fasting blood glucose 100 mg / dL or less, glycated hemoglobin 6.0% or less. All participants recorded age, gender, height, and weight. The questionnaire survey examined smoking history, hypercholesterolemia and hypertension. Hematological tests measured high density lipoprotein cholesterol, low density lipoprotein cholesterol, triglycerides, total cholesterol, fasting blood glucose, glycated hemoglobin, C-reactive protein and serum creatinine. All tests were conducted with the voluntary consent of the participants in accordance with the criteria of the Investigational Review Board (IRB).

(2) SNP analysis

6 ml of blood was collected from all subjects to separate DNA (Genomic DNA). The dried DNA was dissolved in TE buffer (10 mM Tris HCl, pH 8.0, 1 mM EDTA), and the concentration was measured by spectrophotometer and stored at -70 ° C.

Sense primer (5'-TGCCTACTCAGGAGGTGCAG-3 ') and antisense primer (5'-CTTTCAGGGGGTAGAGCCAC-3') were prepared for polymerase chain reaction. The polymerase chain reaction solution was made of 400 ng of genomic DNA extracted from a PCR tube, 10 pM of antisense primer, 10 pM of antisense primer, 2.5 mM of dNTP, 2 units of Taq polymerase, a polymerase chain reaction buffer (10 mM Tris-HCl, 50 mM KCl, 0.1% Triton X-100) was added to make the total reaction solution 20 µl. In the polymerase chain reaction for DNA amplification, the reaction mixture was heated at 94 ° C. for 30 seconds to pre-denaturate the template DNA, followed by denaturation at 94 ° C. for 30 seconds, and annealing at 64 ° C. for 30 seconds. annealing), the DNA was amplified by repeating the extension over 35 times at 72 ° C. for 30 seconds, and the last extension was maintained at 72 ° C. for 10 minutes. Since the reaction solution was subjected to electrophoresis for 30 minutes at 100 V using a 1.0% agarose gel (agarose gel) to confirm the gene.

The gene polymorphism of PSMB5 rs2230087 was read using an Applied Biosystems 7300 Real Time PCR System (Applied Biosystems).

Statistical analysis was performed using the SPSS program. Data is expressed as mean standard deviation. If the p value was less than 0.05, it was defined as statistically significant.

The results of the gene polymorphism readings are shown in Tables 1 and 2 below.

Table 1 . Genotype Frequency of PSMB5 (rs2230087) Polymorphism in Patients with Type 2 Diabetes

Gene Allele Genotype T2DM group
(n = 281) Control group
(n = 146) p value PSMB5

A / G

GG 238 (84.7%) 142 (97.3%) 0.000 GA 12 (4.3%) 3 (2.1%) AA 31 (11.0%) 1 (0.7%)

* Comparisons were performed by Fisher's exact test.

Table 2 . Genotype and Allele Frequency of PSMB5 (rs2230087) Polymorphism and Type 2 Diabetes Risk in Patients with Type 2 Diabetes

PSMB5
(rs2230087) T2DM group
(n = 281) Control group
(n = 146)  p value Crude OR
(95% CI) Adjusted OR
(95% CI) Genotype distribution
GG
GA + AA


238 (84.7%)
43 (15.3%)

142 (97.3%)
4 (2.7%)
0.000



One
6.414
(2.255-18.245)

One
6.821
(2.330-19.969) Allele frequency
G
A

0.868
0.132

0.983
0.017
0.000


One
8.704
(3.478-21.782)
One
9.421
(3.696-24.019)

* Comparisons were performed by Fisher's exact test.

** Adjusted ORs (Odds ratios): adjusting for age and sex by binary logistic regression analysis.

*** CI indicates 95% confidence interval.

As can be seen from Table 1, comparing genotype frequency and risk for PSMB5 (rs2230087) polymorphism in type 2 diabetic patients and the control group, the GA and AA genotypes in the diabetic patients compared to the control group were significantly more frequent than the GG genotype. Is high (p value 0.000).

In addition, as can be seen from Table 2, the genotype (GA + AA) having the G allele and the genotype of the A allele showed a significant increase in the genotype of GA + AA in diabetic patients, and when the relative risk was calculated, GA + The AA genotype had a 6.414 (95% CI 2.255-18.245) higher risk of developing diabetes than the GG genotype, with a higher risk of developing 6.821 (2.330-19.969) times after age and gender correction. A rare variant of the A allele increased significantly in diabetics (p value 0.000), and relative risk calculations showed that people with the A allele had a 8.98 (3.478-) risk of developing diabetes than those with the G allele. 21.782) were higher, and the relative risk was higher at 9.421 (3.696-24.019) after correcting age and gender.

<110> Kyungpook National University Industry-Academic Cooperation Foundation <120> USE OF SNP FOR DIAGNOSING TYPE II DIABETES MELLITUS <130> P07-049-KNU <160> 3 <170> KopatentIn 1.71 <210> 1 <211> 9050 <212> DNA <213> Homo sapiens <220> <221> gene (222) (1) .. (9050) <223> human PSMB5 gene which has sigle nucleotide polymorphisim at          8867th base (G / A) (NCBI refSNP ID: rs2230087), wherein the 8867th          base is described as A. <400> 1 ctttctgccc acactagaca tggcgcttgc cagcgtgttg gagagaccgc taccggtgaa 60 ccagcgcggg tttttcggac ttgggggtcg tgcagatctg ctggatctag gtccagggag 120 tctcagtgat ggtctgagcc tggccgcgcc aggctggggt gtcccagaag agccaggaat 180 cgaaatgctt catggaacaa ccaccctggc cttcaaggtg tggagccagc ccccttgcca 240 ggctgagtac tgaacgcccg cgacttgcct ggcctcccag cctgaccgga gtttgcggtg 300 gccttggcca ccaacccagg cggaggctgg gtcctctcgt tcggaccgca gcagttttgc 360 tgtctcattg gcccaggagg cccctgtctt tgctctggtt ggaaagggtg gcggtggggc 420 cgggggtaag tcgggggagg aaggtgaggc ctcttgggtt gattcctagg cgaccccagg 480 ttgctgctgt tcaggatctt ctagctggga aaggccgcaa caacaatgaa tcttggccga 540 gggaaggcta ttgtaaacgt ttaccaaagt tttggaaact gggcggcccc tcaaggcagg 600 ggtaactgaa actggcatga aggggaagcg tggtgagggt aagcattgta ctaagaatca 660 gacatttggg gttccttgcc cagctctgca ataactagct ctagattgag aaatcatttg 720 atttcattga acctgtttcc tcatctataa aatgatttgt tttttttttt tttttgaaac 780 agggcttttc gctgtcgctc aggctggagt gcagtggctc cagcgcctca gcctccagag 840 tagctgacac tataggcaca caccaccacg acaggctaag tttttttact ttttgtagag 900 atggatctcc ctgtgttgcc caggctggtc tccagcacct ggcctcaagc ggtcctcccg 960 ccttggtctc ccaaaatgcc tgggttacag gagtgagcca ccacgcccat cctataaaat 1020 gaatttaata tgggttatat ccatgtcaag ggattgatgt gaagctcggg tgacattaat 1080 atatggtgaa gtatagaagt atactgtgaa gtataaaagg acggagaaag agatgtgctg 1140 ggtttggatt gatgcaaaaa gaagtatgta ggagtgtttt tgtggtctta tgtggcctgt 1200 tttgtgtttt cctctgatct taacagttcc gccatggagt catagttgca gctgactcca 1260 gggctacagc gggtgcttac attgcctccc agacggtgaa gaaggtgata gagatcaacc 1320 catacctgct aggcaccatg gctgggggcg cagcggattg cagcttctgg gaacggctgt 1380 tggctcggca atgtcgaatc tatgagcttc gaaataagga acgcatctct gtagcagctg 1440 cctccaaact gcttgccaac atggtgtatc agtacaaagg catggggctg tccatgggca 1500 ccatgatctg tggctgggat aagagaggcc ctggtgagtt aagctgcaac cacatgatcc 1560 ttgggctgac actacagtga ggaggggttg gatggacaga tgaattaaag ggcttggtgt 1620 caatgctgaa gaagtgtaat taggtcctta gcccttcctt ctctcttttt ttttttcctg 1680 tttttttaga ctgtgtctca ttctgtcgct caggctggag tgcagtggca tgatctcagc 1740 tcactgcagc ctccgccccc aggttcagga attcaagacc agcctggcca acatggcgaa 1800 accccatctc tactaaaaat acaaaaatta gcccggcgtg gtggcgggcg cctgtaatac 1860 cagctgctca ggaggctgag gcatgagaat tgcttgaacc caggaggtgg atgttgcagt 1920 gagcagagat cgcgccactg cactccagcc tgggcgacag agtgagactc tgtctcaaaa 1980 aaaagaattt aagaattcgt tgaaggccgg gcgtggtggc tcactcctgt aatcccagca 2040 ctttgggagg ctgagacagt ggatcacgag gtcaggagat cgagaccatc ctggctaaca 2100 aggtgaaacc ccgtctctac caaaaataca aaaaattagc caggcgtggt ggcaggcgcc 2160 tgtagtccca gctactcagg aggctgaggc aggagaatag tgtgaaccca ggaggcggag 2220 cttgcagtga gtcgagatgg tgccactgca ctccagcctg ggcaacagag cgagactctg 2280 tctcaaaaaa aaaaacaaaa aaaagaattc gttgaatccc acatgtggca gtagtgctat 2340 ttcaagtgtt cattgaccat gtgtggcttg tggctactgt attggactct aatatatggt 2400 tgtccctcaa aagttagtca tttctcttct gtttttttct ttttttcttt ttttgagatg 2460 caaagtttca ctccgatcgt ccagtctggc gtgtaatggc attgtttcag ctcactacaa 2520 cctctgcctc ctgggttcaa gtgattctcc tgccgcagtc tcccaagtac ctgggattac 2580 aagcgcccac cagcatgccc agctaatttt atatttttag tagagatggg gtttcaccat 2640 gttggccagg ctggtctcga aatcctgacc tcaggtgatt cacccgcctc agcctcccaa 2700 agtgccagga ttaaaggcgt gagccgctgc acccggcctc ttctcttttt ttctttagag 2760 agtctttggc ctagaagggg ccttttgggc tcctctcttt tttgaggggg tggggatgga 2820 gtctcgctct gtcgccccga ctggagtgca gcggcgggat ctcagctcac tgcaatctct 2880 gcctcccagg ttcaagtgat tctcctgcct caaccttcca tgtagctgag actacaggca 2940 tatgccacca tgcgcgtcta atttttgtat ttttagtaga gacagggttt caccatgttg 3000 gccaggctgg tcttgaactc ctgacctcag gtgatctgcc cacctcagcc tgccaaagtg 3060 ttgagattac aggagtgagc cattgcgcct agtcctgggc tcatttcttt tatttctttg 3120 ctggttataa tcacccttac aatcatgcta ggtccggttc tatcatctct gaaaataagt 3180 tccttagcct gtttttcctt tccagcattt aagaaacttg ccagacagtt caactttgta 3240 tttaatgtca tttcctcatc agccagtttt gaattgcccg tttagtaaag ttggagtata 3300 gctagtcacc ttctaatttt cccttccata tgcttaaaaa taattattct ttcctccttt 3360 tgattttgct tcattcattt acctgcttac taaataccta gttatcttat agaacaatta 3420 tagggtacct accacttttt cacttgcttt caaacgagaa tagaaagggc tgtagggagt 3480 atccacttac tacccaatgg aggatgaaag gaaattaaaa accggatgtg agagaggagc 3540 tcactttgtt actttggaaa ttgagatgct tttctctgaa aggtgggaaa agacagttcc 3600 atacatttta cgtgatccag ggaagcattt tggaaatgtt gctttgaacc tttagctggc 3660 ggatttgttc taggtgatgg aaacagtgta aaagagtaca gggaccgggt aacaaggaaa 3720 atccagggag actgagcatc atttaaacag tgccttcttc aagaggcctt cctgatgacc 3780 ctgtcccatt cattgtgagt cagccccttg cctcatatta cagttggccc ttgagcaaca 3840 tgggtttgaa tcactcaggt ccacttatac gcaggtattt ttcaacctaa ttctgattga 3900 aaatacacat tcccagaatt tgaaacccaa atatacagag gaccaattta tttatttatt 3960 tatttattta tttttgagat ggagttttcc tcttgtcacc catgctggag tgcaatggcg 4020 cgatctcagt tcattgcaac ctccacctcc tgggttcaag tgatctcctg cctcagcctc 4080 ccgagtagct gggattacag acacccgcca ccctgccagc taacttttgt atttttagta 4140 gagacagggt ttcaccatgt tggccaggct ggtcttaaac tcctgacctc gggtgatcca 4200 cctgcctcag cctcccaaag tgctgggtgg cgtgagccac catatctggc caacttttca 4260 tatatgcagg ttctgcaggg ccaactttgg gacttgagtg tgcatagatt ttggtatatg 4320 caggggtctt gggacaaatc cccttcatat accaaaggat gactagttaa cccaaggctg 4380 aataagataa ataacctttt ttttttttta ttgagacgga gtctcgctct gtcgcccagg 4440 ctggagtgca gtggtgccat ctctgctcac tgcaagctcc gcctcccgga ttcacgccat 4500 tctcctgcct cagcctcccg agtagctggg actacaggtg cccgccacca cgcccggcta 4560 attttttgta tttttagtag agacggggtt tcactgtgtt agccaggatg gtctcaatct 4620 cctgacctca tgatccaccc gcctcggcct cccaaagtgc tgggattgca ggcgtgagcc 4680 acagctcctg gcctgataaa taccgttttt aaaagaacca gttggctggg catggtggct 4740 catgcctgta atctcagcac tttgggaggc cgagatgggt ggatcaccgg agatcaggag 4800 ttcgagacca gcctgaccaa catggtggaa ccttgtctct actaaaaata caaaattatt 4860 gggaggccga ggcgggcgga tcacgaggcc aggagatcga gaccatcctg gctaacacgg 4920 tgaaacccca tctctactaa aaatacaaaa aattagctgg gcgtggtggt ggtcgcctgt 4980 agtcccagct acttgggagg ctgaggcagg agaatggcgt gaacccagga ggtggagctt 5040 gcagtgagct gagatcgtgc cactgcactc cagactgggc aacagagcaa gactccatct 5100 caaaaaaaaa aaatacaaaa ttagccggct gtggtggcac atgcctgtaa tcccagctac 5160 tcagggagat gaggcaggag aatctcttga accctggagg tgcatgcctg taatcccagc 5220 tactcaggag gctgaggcag gagaatcaat cacttgaacc tgggaggcag aggttgcagt 5280 aagctgagat tgtgccattg cactccagct tgggaaacaa gaaagaaact ttgtctcaaa 5340 aatcagtcag tcaatcaata aataaaaaga accagtcaaa tgcggaagga gttttgagga 5400 ggaagagatt acttatgatg ggaggaattg gggaagactt catggattgg tggtggttgt 5460 cctggatgta aaaaagggaa agggaacata acaaagaaag ggaggaaaga agcatgagct 5520 tttgaggagt aactgaaagt ccagtttaat tggaactaag tgggtgaagg ggaatagtga 5580 gccataaagc cagaagggta ctttggaacc agatgatgac ccccctctag gcaagagagc 5640 atggcactgg ggctgaggag gggccagaga aaggagtaag gtagacatca gaatcacctg 5700 aacctgaaaa ccaataaggg cagaggtgct caagctggct tgcattaaaa tcacctgggg 5760 gacttaaaaa acaagttcca tacctgtttc ccaccccaca ccagttataa aaaataccta 5820 gggtaagtcc tgtgcatttt tgaaaaaaat ctttgttaga tgactctagt gcacagtgag 5880 atgtgagaat cactgaaaca ggaaaaagag tagtgatgct attggcagaa tttcagaaga 5940 caggaggaag cgtgggtttg tggcgcacaa gagagatgag tgtggtaggt aatggagaac 6000 cactggtatc taagtttagg aagtttgaat ctgtacatga tgacaaagac atccagctgg 6060 agggatccag taggcagttg ggattgaaca ctttgtctaa attttcatct gtaccaaaag 6120 tcagctagtt ttttcataaa gggccaaata gtaaatattt taggtctgca ggcctaaatc 6180 tttgtctcaa atacttaact ctgccggtgt tgcacaaaac aaccatagac agtatggaaa 6240 cagtggctct gttccaataa cattttaatt acaaaaataa aggcagttgg cctggccaaa 6300 ccctggtcta taccagcaat agaaatataa tgtgagccat atatgtaatt ttacatttta 6360 tactagctac gcttttaaaa ttaaaaaaaa ttaatttcaa tatattttgt ttaacctaat 6420 aaatccaaaa tattatgtta gcaggtagta ctagttcatt tcaaatgctc aactagtcct 6480 gtgtggctag tggctaccat atggacagtg caggtcttta caatgtgatt acaaactcta 6540 ggctttatta tcctcctgtg taaagtatgt cttgttctcc agatagtcta aactctttga 6600 gttctggcat tatggagttt gtttttctgt cacacaggct gcagtgcagt ggcataatca 6660 cagctcactg tagcctcaac ctcctgggct caagtgactg ggattatggg gtcttttgat 6720 gggggagtgg ggattatgtt ttgtttgttt gttttttgag acagagtttc actcttgttg 6780 cccaggctgg agtgcagtgg tgcgatctcg gctcactgca aactccacct cccaggttca 6840 agtgattctc ctgcctccgc ctcctgagta gctgggatta caggcatgcg ccaccacgcc 6900 tggctaattt tgtattttta gtagagacag ggtttctcca tgttggtcag gcttgtcttg 6960 aactcctgac ctcaggtgac ccacctgcct cggcctccca aagtgctggg attacgggtt 7020 tgagccactg cgcctggccg gggattatgt tttaaatgtt atctttcaca gtgtctgaag 7080 ttctgtgctt gaaacctaag tcatttggaa tgtacttgtt ttgtgggtgt gctgagagga 7140 tcggcaacat ggcaaggtag ttattataat ataaggtgag atggggtggt atgttgtaga 7200 atcctggagt cttaatatta aattttattc cttcaggttt tttttttttc ctgaagacag 7260 catctcagtc tgttgcccaa gctggagtgt ggtggtgtga ttatagctta ctgcagcctg 7320 caagtcctga gctcaaacaa tcctcctgtt tcaacctccc acagacatgc accaccacac 7380 tcagctagct tttaaaaaaa taataataat tttatagaag tcctatcttg tatttatgtt 7440 ggcttccctg ggtttctctt ttatatatat atatattatt ttatttattt attattatta 7500 ttattatttt tttttttttt gagacaaagt cttgctctgt ctcccaggct ggagtgcagt 7560 ggcgcgatct cggctcaccg caagctccgc ctcccgggtt tacgccattc tcctgcctca 7620 gcctcctgag tagctgggac tgcaggcgac cgcaaccaca cctggctaat tttttgtttt 7680 ttagtagaga cggggtttca ccgtgttagc caggatgatc tcgatctcct gacctcgtga 7740 tccacccgcc tcggcctccc aaagtgctgg gattacaggc gttagccacc gcgcctggcc 7800 tatttattat attattttga gacagcgttt cactcttgtt gcccaggctg gagtgcaatg 7860 gcgcgatctt ggcttaccgc aacctttgcc tcccaggttc aagtgattct cctgcatcag 7920 cctcctgagt agctgggatt ataggcatgt gccaccacgc ctggcaaatt ttgtattttt 7980 agtagagatg gggtttcttc atgttggtca ggctggtctc aaacgcctga cctcaggtga 8040 tctgcccacc tcagcctccc aaagtgctgg gattacaggc atgagccacc gcgcccggcc 8100 tattttttta tttttttaaa ttttattcta ggttttaaga gaactactgc tggccgggcg 8160 aggtggctca cgcctgtaat cccagcactt tgggagtccg agaagggcag atccccagag 8220 gtcaggagtt caagaccagc ctgaccaaca tggtgaaacc ccgtctctac taaaaataca 8280 aaaattagct gggcgtggtt acgagcgctt gtaatcctag ctacctggga aactgaggca 8340 ggagaatcac tttaacccag gaggcggagg ttgtggtgag ccgagattgc gccattgcac 8400 tctagcttgg gcaacaagag caaaactcca tctcaaaaaa aaagaactac tgccttggca 8460 tatgaattag atatcaaaag gggtgatctt tttttttttc ctgctaacct catctccctt 8520 tccaggcctc tactacgtgg acagtgaagg gaaccggatt tcaggggcca ccttctctgt 8580 aggttctggc tctgtgtatg catatggggt catggatcgg ggctattcct atgacctgga 8640 agtggagcag gcctatgatc tggcccgtcg agccatctac caagccacct acagagatgc 8700 ctactcagga ggtgcagtca acctctacca cgtgcgggag gatggctgga tccgagtctc 8760 cagtgacaat gtggctgatc tacatgagaa gtatagtggc tctaccccct gaaagagggt 8820 ggatgcagct gcttgtgttt cttggggtga ctgtcattgg taatacagac acagtgaccc 8880 atcctccatc ctatttatag tggaagggcc ttcaattgta tcagtacttt tttttaagct 8940 ctggcacatt gacctctatg tgttaccagt cattaatgag ctgctgcaga ggtgactatt 9000 tgttttactt tcttggatgt taacattaca ctactcacta ctcaatctca 9050 <210> 2 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> sense primer amplifying PSMB5 <400> 2 tgcctactca ggaggtgcag 20 <210> 3 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> antisense primer amplifying PSMB5 <400> 3 ctttcagggg gtagagccac 20  

Claims (10)

NCBI refSNP ID of the polynucleotide consisting of the nucleotide sequence of SEQ ID NO: 1 Type II diabetes diagnostic polynucleotide or its complementary polynucleotide consisting of 10 or more consecutive bases including the base of the SNP position of rs2230087. A polynucleotide that specifically hybridizes to the polynucleotide of claim 1 or a complementary polynucleotide thereof. The polynucleotide of claim 2, wherein the polynucleotide that specifically hybridizes with the polynucleotide or its complementary polynucleotide is an allele specific probe or an allele specific primer. The polynucleotide of claim 3, wherein the polynucleotide that specifically hybridizes with the polynucleotide or its complementary polynucleotide is a primer having a nucleotide sequence of SEQ ID NO: 2 or SEQ ID NO: 3. A polypeptide encoded by the polynucleotide of claim 1. A composition for diagnosing type 2 diabetes, comprising the polynucleotide of claim 1, a polynucleotide that hybridizes specifically thereto, or a polypeptide specifically encoded by the polynucleotide thereof. A microarray for diagnosing type 2 diabetes, comprising the polynucleotide of claim 1, a polynucleotide that hybridizes specifically thereto, a polypeptide specifically encoded by the polynucleotide thereof, or a cDNA thereof. Type 2 diabetes diagnostic kit comprising the microarray of claim 7. Contacting the polypeptide encoded by the polynucleotide of claim 1 with a test subject; And Determining whether the test subject exhibits an activity that enhances or inhibits the function of the polypeptide. Screening method of medicament for treating type 2 diabetes comprising a. delete