KR20070113586A - Wnt10b snp makers for the prediction of susceptibility to obesity - Google Patents

Abstract

A Wnt10B polymorphism marker is provided to be usefully used for effectively predicting obesity and susceptibility to various obesity-related diseases by using property of -607C>G of a Wnt gene closely related to body fat and abdominal fat area. A Wnt10B polymorphism marker for predicting susceptibility to obesity is characterized in that -607th base from a transcription starting point of a Wnt10B gene(105th base of SEQ ID : NO. 3) is G and it is a polynucleotide consisting of 20-100 consecutive DNA sequence and including the -607 base. A Wnt10B haplotype marker for predicting the obesity susceptibility is characterized in that each of -1347th base, -607th base, -295th base, +121st base, +5555th base or +5957th base from the transcription starting point of the Wnt10B gene is T, G, T, G, C, or A, respectively. A method for predicting susceptibility to obesity comprises the steps of: (a) isolating a genome DNA from a blood sample of an individual; (b) detecting a polymorphic portion in the marker from the isolated genome DNA; and (c) identifying the detected polymorphic portion. A kit for predicting the obesity susceptibility comprises a primer pair capable of amplifying a -607C>G polymorphic portion of the Wnt10B gene.

Description

Wnt10B SNP makers for the prediction of susceptibility to obesity

Figure 1 shows the results of measuring the area of the adipose tissue cross section using CT, (A) is a diagram showing the abdominal fat cross-section, (B) is the abdominal subcutaneous fat area, (C) is a diagram showing the abdominal fat area.

Figure 2 shows the gene map and haplotype of the Wnt10B gene, (A) is polymorphism of Wnt10B, (B) is an association equilibrium constant D 'and r2 between the six SNPs, (C) is a haplotype of Wnt10B Picture.

* SNP indicates primer site for sequencing of Wnt10B genomic DNA of SNP with minor allele frequency shown in parentheses and arrows.

Figure 3A is a diagram showing the relationship between the -607C> G SNP and the total abdominal fat area of Wnt10B, Figure 3B is a diagram showing the relationship between the Wnt10B -607C> G SNP and abdominal subcutaneous fat area.

The present invention relates to a Wnt10B polymorphism marker for predicting the risk of obesity and a method for predicting the risk of the disease using the same. More specifically, it relates to obesity and related by measuring the association of Wnt10B polymorphism with body fat mass and abdominal fat area. The present invention relates to a marker for predicting the risk of disease and a prediction method using the same.

Wnt contains a family of secretory signaling proteins that regulate a variety of developmental processes. Wnt signaling is known to maintain preadipocytes in undifferentiated state through inhibition of the lipogenic transcription factors CEBPA and PPAR-γ (Ross). et al. Science 289: 950-953, 2000). WNT signaling in profat cells is inhibited by overexpression of axin or dominant-negative TCF4 and these cells differentiate into adipocytes. Destruction of WNT signaling also allows myoblasts to transdifferentiate into adipocytes in vitro, indicating that this pathway is important not only for adipocyte differentiation but also for determining the fate of mesenchymal cells. This WNT signaling is known to be mediated by Wnt10B, which encodes a 389 amino acid protein that has the same 96.6% sequence as mouse Wnt10B and is highly synthesized in adult heart and skeletal muscle (Hardiman et al. Cytogenet). Cell Genet. 77: 278-282, 997). In addition, Wnt10B also plays an important role in bone development, by inducing the osteoblast production transcription factors Runx2, Dlx5 and OSX (osterix) to differentiate the cells into osteoblast lineage and osteoblast formation by inhibiting the expression of PPAR-γ. To promote. Finally, Wnt10B − / − mice are endogenous bone formation regulators that reduce osteocalcin in sponges and serum (Bennett et al., Proc Natl Acad Sci US A. 2005 Mar 1; 102 (9): 3324-3329, 2005 )

As described above, Wnt10B inhibits the expression of adipogenic transcription factor C / EBPalpha by inhibiting the expression of adipogenic transcription factor C / EBPalpha and inhibits the apoptosis of bipotential mesenchymal precursors and inhibits the expression of PPAR-γ to produce osteoblasts. plays an important role in promoting osteoblastogenesis.

In order to determine whether the mutation of Wnt10B is involved in human obesity, studies have been conducted to determine whether there is a mutation in the gene in obese patients. Study of 96 UK obesity studies and 115 European Italian obese patients who started under 10 years of age showed that one child with obesity as a child was heterozygous for the C256Y mutation. This is because Wnt10B promotes lipogenesis by eliminating the ability to activate normal WNT signaling. Relatives of promoters with these alleles were overweight or obese, although three other rare missense variants were found in obese progenitors, but they are hard to say that they are clearly related to obesity in family studies, so family tree analysis is one of these variants. And no clear relationship between obesity and obesity (Christodoulides C, et al., Diabetologia. 494: 678-684. 2006).

Accordingly, the present inventors have analyzed the results of genetic analysis and statistical analysis of Korean obese patients, and the -607C> G monobasic polymorphism of Wnt10B and a haplotype including the polymorphism were associated with the risk of obesity, thereby predicting the risk of obesity. Confirmed that it can be used to complete the present invention.

An object of the present invention is to provide a Wnt10B polymorphism marker for predicting obesity risk and a method for predicting obesity using the same.

The present invention provides a Wnt10B polymorphism marker for predicting obesity risk.

In addition, the present invention provides a Wnt10B haplotype for predicting the risk of obesity.

In addition, the present invention provides a method for predicting the risk of obesity by identifying the -607C> G polymorphic site of the Wnt10B gene.

In addition, the present invention provides a kit for predicting the risk of obesity, including a primer capable of amplifying the -607C> G polymorphic site of the Wnt10B gene.

Hereinafter, the present invention will be described in detail.

The present invention provides a Wnt10B polymorphism marker for predicting obesity risk.

The present inventors extracted genomic DNA from 1020 women who visited the obesity clinic and Wnt10B genome sequence (GeneBank Accession No.:AC073610), promoter from the National Center for Biotechnology Information Base sequences for the region (SEQ ID NO: 1) and coding region (SEQ ID NOs: 2 and 5) were obtained.

The inventors performed PCR and sequencing of the Wnt10B promoter and coding region to search for SNPs present in the promoter region and the coding region of the Wnt10B gene (see Table 1). As a result of sequencing of the product obtained by PCR, the Wnt10B SNP region present on the 12q13 chromosome showed that the polymorphic region was -1347A> T, -607C> G, -295C> T, + 121T> G, + 5555T> Six kinds of C, + 5957C> A were identified, and it was confirmed that this is an SNP site which is already known.

As a result of investigating the associations to the SNPs discovered above, since all these SNPs are related and linked together as one SNP, analysis of one SNP can determine the types for the remaining six SNPs. The primers were used to identify the genotype for -607C> G by the SNPshot method.

The present inventors first examined the correlation between -607C> G polymorphism and body type, and found that the polymorphism was not significantly related to body weight, but could be related to weight loss, and was associated with waist hip ratio (WHR) and The body mass index (BMI) had a significant association in the recessive model (see Table 4).

In addition, the correlation between Wnt10B -607C> G polymorphism and body composition showed a close correlation with the decrease of body fat mass and body fat percentage, and no correlation with other components was observed (see Table 5). The fat area was associated with a decrease in total abdominal fat area and subcutaneous fat area, and no significant difference was observed between abdominal visceral fat area (see Table 6 and FIG. 3). No correlation was found between blood lipid components, glutamate oxaloacetate transaminase (GOT), glutamate pyrovate transaminase (GPT), and blood pressure (see Tables 7 and 8).

In summary, the Wnt10B -607C> G, a minor allele of the SNP present in the promoter and coding region of Wnt10B, has a significant correlation with the decrease in body fat mass and abdominal fat area.

In addition, the present invention provides a Wnt10B haplotype for predicting the risk of obesity.

The present inventors examined the maps of the Wnt10B promoter and the SNP positions of the coding sites and LD blocks using these, and confirmed that they were all related to form a haplotype (FIG. 2). As a result of the investigation of obesity related to -607C> G, which is one of the related SNPs, there was a significant relationship between the two. Therefore, the haplotype, which is a set of SNPs, was significantly associated with the decrease of body fat and abdominal fat area. Can be inferred.

In addition, the present invention

1) separating genomic DNA from blood samples of the subject;

2) detecting the polymorphic site in the marker of claim 1 in genomic DNA isolated in step 1; And

3) It provides a method of predicting the risk of obesity, including the step of identifying the polymorphic site detected in step 2.

Step 2) is performed by sequencing analysis, hybridization by microarray, allele specific PCR, dynamic allele-specific hybridization (DASH), PCR extension analysis or TaqMan technique. Can be performed.

In addition, the present invention provides a kit for predicting the risk of obesity, including a primer capable of amplifying the -607C> G polymorphic site of the Wnt10B gene.

The kit comprises primer pairs capable of amplifying the -607C> G polymorphic site of the Wnt10B gene and the primers are preferably provided in SEQ ID NOs: 10 and 11 and Table 2, respectively, in order for the -607C> G polymorphic site, respectively. As a primer, a PCR product using the same may be analyzed to determine the sequence of the -607C> G polymorphic site to predict the risk of obesity. The primer pair is irrelevant in size and can be readily designed by those skilled in the art using conventional primer selection software.

Hereinafter, the present invention will be described in detail by way of examples. However, the following Examples are only for illustrating the present invention and do not limit the present invention.

Example 1 Wnt10B SNP Identification on Chromosome 12q13

<1-1> Selection of Test Subjects and DNA Sampling from Samples

Genomic DNA was extracted from 1020 women who visited the Kirin Oriental Hospital Obesity Clinic (Seoul, South Korea). After acquiring the blood sample of the subject under the approval of the Institutional Review Board of the Korean Institute of Oriental Medicine, it was extracted using a genomic DNA extraction kit (LaboPass ™ Blood kit, COSMO Co., Korea) according to the manufacturer's instructions.

<1-2> PCR analysis and SNP search

Wnt10B Genome Sequence (GeneBank Accession No.:AC073610) from the National Center for Biotechnology Information (NCBI, http://www.ncbi.nlm.nih.gov/), promoter region (SEQ ID NO: 1 Nucleotide sequence for the coding region (SEQ ID NOS: 2 and 5), and PCR of the Wnt10B promoter and coding region in 45 subjects to search for SNPs present in the promoter region and coding region of the Wnt10B gene in Koreans. And sequencing was performed. Primer sequences and positions for this are shown in Table 1.

Table 1: Primer sites and sequences to determine SNPs of Wnt10B promoter and exon sites

part Primer Name order SEQ ID NO: part Promoter and Exxon 1 Pe1_F1 TTT TCC AGC GAG ACC CCA TGT 6 -3155--3175 Pe1_F2 GGG GCC TTC CTG AGA GTC 7 -2544 ~ -2561 Pe1_F3 TGG TTT GGC TCT TGT CCT CTA 8 -1382-1402 Pe1_R2 GGG GTC ACT GTT GGA TAG C 9 -1150 ~ -1168 Pe1_F4 TTC GCT GCC ACT GGG TCT C 10  -832--850 Pe1_R1 GGG GGC ACT CCT TTA TTC TCT G 11 458-479 Exon 2,3 e2-F1 GGG GTG TTT CTT CTT GTT TTG 12 1044-1064 e2-R1 ACG CAG GTG GAA GTT AGG C 13 1851-1869 Exon 4 e4-F1 GCT TTT TGT ACC CTT CCT TGT 14 3262 ~ 3282 e4-R1 TGG GTG ACT TGC TGA TGG TGA G 15 3920 ~ 3941 Exon 5 e5-F1 TGT CTT CAA TTC TGC CAT CAT A 16 5011-5032 e5-R2 GCC CAG GCT GCA GTA AAG GAG 17 6109 ~ 6129

PCR amplification conditions were prepared by adding genomic DNA, Taq polymerase, buffer to the primer pairs described above using DNA polymerase (Taq polymerase, Takara), and denatured at 94 ° C. for 5 minutes. The reaction was completed for 30 minutes at 94 ° C., 1 minute at 60 ° C., and 4 minutes at 72 ° C., followed by extension at 72 ° C. for 10 minutes.

The resulting product was electrophoresed on a 1.5% agarose gel to confirm their size, and a sequencing kit (DynamicTM ET Dye Terminator Kit, Amersham Biosciences, UK) and a genetic analyzer (ABI 377 DNA Sequencers, Global Medical Instrumentation Inc) , USA) was used to perform sequencing according to the manufacturer's instructions.

Then, using potential alignment software (ClustalX alignment software, Ver 1, http://bips.u-strasbg.fr/fr/Documentation/ClustalX/), the potential heterozygote positions were marked and present on the 12q13 chromosome. Wnt10B SNP polymorphic sites were identified as -1347A> T, -607C> G, -295C> T, + 121T> G, + 5555T> C, and + 5957C> A. The location and type of each SNP is shown in Table 2 and the sequencing analysis identified the SNPs as known SNPs.

Table 2: SNP site of Wnt10B promoter or exon site in 45 Korean women

Sample ID Seat -1347 -607 -295 +121 +5555 +5957 One AA GG CC TT TT CC 2 AA GG CC TT TT CC 3 AA GG CC TT TT CC 4 AT CG CT GT CT CA 5 AT CG CT GT CT CA 6 TT CC TT GG CC AA 7 AT CG CT GT CT CA 8 AA GG CC TT TT CC 9 AT CG CT GT CT CA 10 AA GG CC TT TT CC 11 AT CG CT GT CT CA 12 AA GG CC TT TT CC 13 AT CG CT GT CT CA 14 TT CC TT GG CC AA 15 AT CG CT GT CT CA 16 AT CG CT GT CT CA 17 AT CG CT GT CT CA 18 AA GG CC TT TT CC 19 AT CG CT GT CT CA 20 AA GG CC TT TT CC 21 AT CG CT GT CT CA 22 AT CG CT GT CT CA 23 AA GG CC TT TT CC 24 AT CG CT GT CT CA 25 AT CG CT GT CT AC 26 AA GG CC TT TT CC 27 AT CG CT GT CT AC 28 AT CG CT GT CT AC 29 AT CG CT GT CT AC 30 AT CG CT GT CT AC 31 AT CG CT GT CT AC 32 AT CG CT GT CT AC 33 TT CC TT GG CC AA 34 AA GG CC TT TT CC 35 TT CC TT GG CC AA 36 AA GG CC TT CT AC 37 TT CC TT GG CC AA 38 AT CG CT GT CT AC 39 AA GG CC TT TT CC 40 AT CG CT GT CT AC 41 AT CG CT GT CT AC 42 AA GG CC TT TT CC 43 AT CG CT GT CT AC 44 AT CG CT GT CT AC 45 AA GG CC TT TT CC

<Example 1-3> Location and Relationship between SNPs

Figure 2 shows the association between the Wnt10B SNP polymorphism site and each of the SNPs present on the 12q13 chromosome of Koreans. The Hardy-Weinberg equilibrium was calculated to determine equilibrium at each position in the population between individuals, and ┃D'┃ and r ² to measure the association disparity of all pairs that could occur between the two alleles. (Hedric, PW, Genetics , 117: 331-341, 1987). As a result, the value of associative unbalance coefficient (계수 D'┃) between -1347A> T, -607C> G, -295C> T, + 121T> G, + 5555T> C, and + 5957C> A was 1, and r ² The value was also very high, 0.995-1, meaning that all six SNPs were related.

Example 2 SNP Identification of Research Subjects

Since all the SNPs searched for in the above example were related, 1020 subjects of Example <1-1> were representatively identified for -607C> G SNP. Primers used for -607C> G SNP identification were shown in Table 3, and SNP identification was performed according to the manufacturer's instructions using ABI PRISM SNaPshot ™ Multiplex Kit (Foster City, CA, U.S.A.).

Table 3: Primer sequences for PCR reaction of -607C> G SNP genotype of Wnt10B

primer order PCR Forward direction GAGCACTTTTATGGCACAAATGA (SEQ ID NO: 18) Reverse TATAGTAGACAACATACGACCACGCTAA (SEQ ID NO: 19) Genotyping GCTAAACCCAAATACATCTXCCAGGAT

Example 3 Characterization of Research Subjects

The present inventors obtained the blood in Example <1-1> 1020 General characteristics of the study subjects are shown in Table 4. Specifically, age, height and weight were measured, and body fat mass was measured using a commercially available bio-AC resistance analyzer (Inbody 2.0, Biospace Co., Korea). Body mass index (BMI) and waist hip ratio (WHR) were calculated from the measured height and weight. In this case, the body mass index refers to a value obtained by dividing the mass (kg) by the square of the height (m), and the hip pelvic ratio means a value obtained by dividing the waist circumference by the pelvic circumference. Abdominal body fat area was characterized using computerized tomography (CT) cross sectional picture (Hispeed CT / e, GE, US). General physical characteristics of the study are shown in Table 4 below. All measurements are expressed as mean ± standard deviation (SD).

Table 4: General physical characteristics of women studied

Indicators Mean ± S.D. Age (years) 28.27 ± 8.14 Weight (kg) 66.49 ± 11.53 Height (cm) 160.73 ± 5.41 WHR 0.8796 ± 0.0655 BMI (kg / m ² ) 25.72 ± 4.07

The age-adjusted univariate analysis of the deviations is a linear model to determine the individual correlations between Wnt10b SNP and body composition, abdominal fat area, abdominal subcutaneous fat area, HDL-cholesterol, and total cholesterol indices, depending on the variables to be performed below. (general linear model) was used. Statistical significance was established at the level of p <0.05 and all analyzes were performed using statistical analysis software (SPSS10.1, SPSS Korea, Seoul, Korea).

Example 4 Investigation of Correlation between Wnt10B-607C G Polymorphism and Body Type

The present inventors investigated the correlation between -607C> G and body type, but it was not significantly related to body weight, but it was likely to be related to weight loss, waist hip ratio (WHR) and body mass index (Body Mass Index). , BMI) was significantly associated with the recessive model (Table 5).

Table 5: Correlation analysis between Wnt10B -607C> G polymorphism and constitution in Korean women

Phenotype C / C C / G G / G p value Co-dominance dominant zeal weight 264 (66.60 ± 11.58) 509 (67.00 ± 12.24) 256 (65.35 ± 9.87) 0.195 0.849 0.079 WHR 264 (0.8787 ± 0.0621) 509 (0.8852 ± 0.0705) 256 (0.8695 ± 0.0570) 0.021 0.824 0.011 BMI 264 (25.69 ± 3.82) 509 (25.99 ± 4.40) 256 (25.23 ± 3.56) 0.079 0.905 0.038

Example 5 Correlation between Wnt10B-607C G Polymorphism and Body Composition

We investigated the correlation between Wnt10B-607C> G polymorphism and body composition. Body composition of the test subjects was measured using Inbody 2.0 (Biospace Co., Seoul, Kore).

As a result, Wnt10B-607C> G was closely related to the decrease in body fat mass and body fat percentage, and no association with other components was observed (FIG. 5).

Table 6: Relationship between Wnt10B -607C> G polymorphism and body composition in Korean women

Phenotype C / C C / G G / G p value Co-dominance dominant zeal FAT_MASS 264 (23.26 ± 7.27) 509 (23.86 ± 8.12) 256 (22.41 ± 6.40) 0.046 0.843 0.024 Total_Body_Water 264 (30.96 ± 4.28) 509 (30.82 ± 3.93) 256 (30.66 ± 3.59) 0.72 0.485 0.533 Fat_Free_Mass 264 (43.42 ± 5.74) 509 (43.17 ± 5.30) 256 (43.08 ± 4.85) 0.745 0.448 0.717 Percent_Body_FAT 264 (34.25 ± 5.67) 509 (34.81 ± 5.82) 256 (33.69 ± 4.96) 0.032 0.648 0.023

Example 6 Correlation between Wnt10B-607C G Polymorphism and Abdominal Fat Area

We investigated the correlation between Wnt10B-607C> G and abdominal fat area. Body fat area of the test subject was measured using CT.

As a result, Wnt10B-607C> G was associated with a decrease in total abdominal fat area and subcutaneous fat area, and no significant difference was observed between abdominal visceral fat area (Table 7 and FIG. 4).

Table 7: Relationship between Wnt10B -607C> G polymorphism and abdominal fat area in Korean women

Phenotype C / C C / G G / G p value Co-dominance dominant  zeal Abdominal Fat Area (mm ² ) 263 (28559.1 ± 729.6) 504 (29255.5 ± 575.0) 253 (26576.6 ± 646.6) 0.026 0.796 0.009 Abdominal visceral fat area (mm ² ) 263 (5930.2 ± 222.7) 504 (6245.81 ± 80.0) 253 (5641.8 ± 194.1) 0.291 0.74 0.182 Abdominal subcutaneous fat area (mm ² ) 263 (22540.0 ± 584.4) 504 (23009.7 ± 472.0) 253 (20934.7 ± 530.3) 0.020 0.775 0.007

* The number of patients (mean SE) and P values of the age standardized general linear model are shown.

P values less than 0.05 are shown in bold.

* C / C, C / G, and G / G represent homozygotes of common alleles, heterozygotes, and rare alleles, respectively.

Example 7 Correlation between Wnt10B-607C G Polymorphism and Blood Components

The inventors investigated the correlation between Wnt10B-607C> G polymorphism and blood components. In the blood lipid analysis, blood was first collected from a subject fasted for 12 hours or longer, and the collected blood was centrifuged at 2,000 rpm for 10 minutes to obtain a supernatant, and again only serum was obtained. Blood components were measured using an auto-biochemical analyzer (SP-4410, Arkray Co., Japan).

The correlation between Wnt10B-607C> G, blood lipid components, glutamate oxaloacetate transaminase (GOT), and glutamate pyruvate transaminase (GPT) was found to be unrelated to all SNPs (Table 8).

Table 8: Relationship between Wnt10B -607C> G polymorphism and serum chemical profile in Korean women

Phenotype C / C C / G G / G p value Co-dominance dominant zeal GOT 258 (16.74 ± 12.68) 503 (16.83 ± 16.47) 251 (18.18 ± 16.62) 0.457 0.637 0.211 GPT 126 (20.46 ± 22.75) 253 (19.41 ± 19.54) 118 (20.44 ± 30.78) 0.883 0.775 0.768 GLU 257 (107.20 ± 31.36) 500 (110.93 ± 36.63) 252 (103.18 ± 29.39) 0.018 0.658 0.013 HDL_C 226 (57.99 ± 27.01) 436 (58.31 ± 25.02) 228 (56.35 ± 21.97) 0.659 0.846 0.365 T_CHO 255 (177.79 ± 71.65) 498 (174.89 ± 62.24) 253 (174.54 ± 59.14) 0.794 0.497 0.855 TG 258 (80.61 ± 49.67) 501 (81.19 ± 50.04) 253 (84.57 ± 49.04) 0.461 0.643 0.213

Example 8 Investigation of Correlation between Wnt10B-607C G Polymorphism and Blood Pressure

The present inventors examined the correlation between Wnt10B-607C> G and blood pressure, and no correlation was observed (Table 9).

Table 9: Relationship between Wnt10B -607C> G polymorphism and blood pressure in Korean women

Phenotype C / C C / G G / G p value Co-dominance dominant zeal SBP 264 (114.83 ± 13.94) 509 (117.18 ± 47.68) 256 (114.02 ± 12.40) 0.483 0.611 0.392 DBP 264 (71.71 ± 10.01) 509 (72.24 ± 11.34) 256 (72.22 ± 10.10) 0.766 0.515 0.596

In summary, the SNPs present in the promoter and coding region of Wnt10B are related to each other to form a block, and Wnt10B-607C> G, which is a minor allele of these SNPs, reduces body fat and abdomen. It was confirmed that there is a significant correlation with fat area reduction.

As described above, since the -607C> G SNP of the Wnt10B gene according to the present invention is closely related to the body fat mass and the abdominal fat area, their association is useful for effectively predicting the polymorphism of the Wnt10B gene and the risk of developing obesity and related diseases. Can be used.

<110> Korea Institute of Oriental Medicine <120> Wnt10B SNP makers for the prediction of susceptibility to          obesity <160> 19 <170> KopatentIn 1.71 <210> 1 <211> 306 <212> DNA <213> WNT10B promoter <400> 1 ggggctgcag ctccgtcagc ccggcagagc caccctgagc tcggtgagag caaagccaga 60 gcccccagtc ctttgctcgc cggcttgcta gctctctcga tcactccctc ccttcctccc 120 tcccttcctc ccggcggccg cggcggcgct ggggaagcgg tgaagaggag tggcccggcc 180 ctggaagaat gcggctctga caaggggaca gaacccagcg cagtctcccc acggtttaag 240 cagcactagt gaagcccagg caacccaacc gtgcctgtct cggaccccgc acccaaacca 300 ctggag 306 <210> 2 <211> 74 <212> DNA <213> WNT10B exon 1 <400> 2 atgctggagg agccccggcc gcggcctccg ccctcgggcc tcgcgggtct cctgttcctg 60 gcgttgtgca gtcg 74 <210> 3 <211> 263 <212> DNA <213> WNT10B exon 2 <400> 3 ggctctaagc aatgagattc tgggcctgaa gttgcctggc gagccgccgc tgacggccaa 60 caccgtgtgc ttgacgctgt ccggcctgag caagcggcag ctaggcctgt gcctgcgcaa 120 ccccgacgtg acggcgtccg cgcttcaggg tctgcacatc gcggtccacg agtgtcagca 180 ccagctgcgc gaccagcgct ggaactgctc cgcgcttgag ggcggcggcc gcctgccgca 240 ccacagcgcc atcctcaagc gcg 263 <210> 4 <211> 374 <212> DNA <213> WNT10B exon3 <400> 4 gtttccgaga aagtgctttt tccttctcca tgctggctgc tggggtcatg cacgcagtag 60 ccacggcctg cagcctgggc aagctggtga gctgtggctg tggctggaag ggcagtggtg 120 agcaggatcg gctgagggcc aaactgctgc agctgcaggc actgtcccga ggcaagagtt 180 tcccccactc tctgcccagc cctggccctg gctcaagccc cagccctggc ccccaggaca 240 catgggaatg gggtggctgt aaccatgaca tggactttgg agagaagttc tctcgggatt 300 tcttggattc cagggaagct ccccgggaca tccaggcacg aatgcgaatc cacaacaaca 360 gggtggggcg ccag 374 <210> 5 <211> 1213 <212> DNA <213> WNT10B exon4 <400> 5 gtggtaactg aaaacctgaa gcggaaatgc aagtgtcatg gcacatcagg cagctgccag 60 ttcaagacat gctggagggc ggccccagag ttccgggcag tgggggcggc gttgagggag 120 cggctgggcc gggccatctt cattgatacc cacaaccgca attctggagc cttccagccc 180 cgtctgcgtc cccgtcgcct ctcaggagag ctggtctact ttgagaagtc tcctgacttc 240 tgtgagcgag accccactat gggctcccca gggacaaggg gccgggcctg caacaagacc 300 agccgcctgt tggatggctg tggcagcctg tgctgtggcc gtgggcacaa cgtgctccgg 360 cagacacgag ttgagcgctg ccattgccgc ttccactggt gctgctatgt gctgtgtgat 420 gagtgcaagg ttacagagtg ggtgaatgtg tgtaagtgag ggtcagcctt accttggggc 480 tggggaagag gactgtgtga gaggggcgcc ttttcagccc tttgctctga tttccttcca 540 aggtcactct tggtccctgg aagcttaaag tatctacctg gaaacagctt taggggtggt 600 gggggtcagg tggactctgg gatgtgtagc cttctcccca acaattggag ggtcttgagg 660 ggaagctgcc acccctcttc tgctccttag acacctgaat ggactaagat gaaatgcact 720 gtattgctcc tcccacttct caactccaga gcccctttaa ccctgattca tactcctttt 780 ggctggggag tccctatagt ttcaccactc ctctcccttg agggataacc ccaggcactg 840 tttggagcca taagatctgt atctagaaag agatcaccca ctcctatgta ctatccccaa 900 actcctttac tgcagcctgg gctccctctt gtgggataat gggagacagt ggtagagagg 960 tttttcttgg gaaagagaca gagtgctgag gggcactctc ccctgaatcc tcagagagtt 1020 gtctgtccag gcccttaggg aagttgtctc cttccattca gatgttaatg gggaccctcc 1080 aaaggaaggg gttttcccat gactcttgga gcctcttttt ccttcttcag caggaagggt 1140 gggaagggat aatttatcat actgagactt gttcttggtt cctgtttgaa actaaaataa 1200 attaagttac tgg 1213 <210> 6 <211> 21 <212> DNA <213> Pe1_F1 <400> 6 ttttccagcg agaccccatg t 21 <210> 7 <211> 18 <212> DNA <213> Pe1_F2 <400> 7 ggggccttcc tgagagtc 18 <210> 8 <211> 21 <212> DNA <213> Pe1_F3 <400> 8 tggtttggct cttgtcctct a 21 <210> 9 <211> 19 <212> DNA <213> Pe1_R2 <400> 9 ggggtcactg ttggatagc 19 <210> 10 <211> 19 <212> DNA <213> Pe1_F4 <400> 10 ttcgctgcca ctgggtctc 19 <210> 11 <211> 22 <212> DNA <213> Pe1_R1 <400> 11 gggggcactc ctttattctc tg 22 <210> 12 <211> 21 <212> DNA <213> e2-F1 <400> 12 ggggtgtttc ttcttgtttt g 21 <210> 13 <211> 19 <212> DNA <213> e2-R1 <400> 13 acgcaggtgg aagttaggc 19 <210> 14 <211> 21 <212> DNA <213> e4-F1 <400> 14 gctttttgta cccttccttg t 21 <210> 15 <211> 22 <212> DNA <213> e4-R1 <400> 15 tgggtgactt gctgatggtg ag 22 <210> 16 <211> 22 <212> DNA <213> e5-F1 <400> 16 tgtcttcaat tctgccatca ta 22 <210> 17 <211> 21 <212> DNA <213> e5-R2 <400> 17 gcccaggctg cagtaaagga g 21 <210> 18 <211> 23 <212> DNA <213> Wnt10B genotyping primer <400> 18 gagcactttt atggcacaaa tga 23 <210> 19 <211> 28 <212> DNA <213> Wnt10B genotyping primer <400> 19 tatagtagac aacatacgac cacgctaa 28

Claims (8)

Wnt10B polymorphic marker for predicting obesity risk. The method of claim 1, wherein the marker is composed of 20-100 consecutive DNA sequences comprising -607 base G and -607th base (105 base of SEQ ID NO: 3) from the transcription start point of the Wnt10B gene Wnt10B polymorphic marker, characterized in that the polynucleotide. Wnt10B haplotype marker for predicting obesity risk. 4. The marker of claim 3, wherein the marker is -1347th base T, -607th base G, -295th base T, + 121th base G, + 5555th base C from the start point of transcription of the Wnt10B gene, or A haplotype marker, characterized in that the + 5957th base is A. 1) separating genomic DNA from blood samples of the subject; 2) detecting the polymorphic site in the marker of claim 1 in genomic DNA isolated in step 1; And 3) A method of predicting the risk of obesity comprising identifying the polymorphic site detected in step 2. The method of claim 5, wherein step 2) comprises sequencing analysis, hybridization by microarray, allele specific PCR, dynamic allele-specific hybridization (DASH), PCR extension Obesity risk prediction method characterized in that performed by the analysis or TaqMan technique. A kit for predicting obesity risk including a primer pair capable of amplifying a -607C> G polymorphic site of the Wnt10B gene. 8. The kit for predicting obesity risk according to claim 7, wherein the primers are primers having SEQ ID NOs: 10 and 11 or SEQ ID NOs: 18 and 19, respectively, in order for the -607C> G polymorphism site. 9.

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