ADIPOQ gene SNP molecular marker related to pig backfat thickness character and application of ADIPOQ gene SNP molecular marker

Abstract

The invention belongs to the field of molecular biology, and discloses an SNP (Single Nucleotide Polymorphism) molecular marker related to a pig backfat thickness character and application thereof. According to the invention, an SNP (Single Nucleotide Polymorphism) site g.348Agt related to the back fat thickness character of the pig is screened in an intron region of the ADIPOQ gene; the SNP site is located at the 348bp nucleotide at the downstream of the ADIPOQ gene transcription start site, and the polymorphism of the SNP site is A or G. According to the present invention, the pig backfat thickness condition is determined according to the genotype of the mutation site, such that the functional gene and the molecular genetic marker related to the pig backfat thickness are obtained, and the dominant allele of the SNP molecular marker is optimized and the site is rapidly detected so as to determine the pig backfat thickness, reference basis can be provided for accurate evaluation of pig fat deposition capacity regulation related SNP molecular marker assisted breeding, and meanwhile, the method has important significance on pig strain or variety improvement.

Description

Technical Field

[0001] The invention belongs to the field of molecular biology and relates to an ADIPOQ gene SNP molecular marker related to pig backfat thickness trait and application thereof. Background Art

[0002] Pork accounts for 35% of global meat consumption and is an important part of the human diet. Excessive fat deposition in pigs leads to poor feed utilization and is not favored by consumers. It is well known that carcass lean meat percentage is strongly negatively correlated with carcass backfat thickness. In order to align production with consumer demand, one of the key goals of pig breeding programs is to reduce carcass fat and improve growth efficiency and lean meat content. Backfat thickness is a good indicator of indirect prediction of body fat content and has a high heritability, so breeding work attaches great importance to the selection of backfat thickness trait.

[0003] Single nucleotide polymorphism (SNP) of genes has important value and significance in the research and application of gene function. As a high-throughput and high-density genetic marker, SNP is widely used in modern breeding, genetic analysis and other fields. It is a very important molecular marker. It is densely distributed throughout the genome and can fully cover the genome, providing a large number of genetic markers for genome selection, thereby improving the accuracy of breeding. Through the association analysis between SNP markers and traits, genotype individuals that can guide breeding practices can be obtained. Summary of the invention

[0004] The purpose of the present invention is to provide an ADIPOQ gene SNP molecular marker related to the pig backfat thickness trait and its application, and to use the SNP molecular marker to conveniently and quickly determine the pig carcass backfat thickness and carcass lean meat percentage, thereby providing a basis for breeding pigs with high lean meat percentage.

[0005] The purpose of the present invention is achieved through the following technical solutions:

[0006] In the first aspect, the present invention claims the use of a substance for detecting the polymorphism or genotype of the SNP site g.348A>G in the ADIPOQ gene on the pig genome in any of the following:

[0007] (a1) Application in identifying or assisting in identifying pig backfat thickness;

[0008] (a2) Use in the preparation of a product for identifying or assisting in identifying pig backfat thickness;

[0009] (a3) Application in selecting or assisting in selecting pig breeds with low backfat thickness;

[0010] (a4) Use in the preparation of products for selecting or assisting in selecting pig breeds with low backfat thickness;

[0011] (a5) Application in assisting the selection of high lean meat rate pig breeds;

[0012] The SNP site g.348A>G is located at the 348 bp nucleotide downstream of the transcription start site in the SEQ ID NO.2 sequence of the pig ADIPOQ gene, and its polymorphism is A or G.

[0013] The SNP site screened by the present invention is located 348 bp downstream of the transcription start site of the ADIPOQ gene (intron region), the genotype of the SNP site is AA, AG or GG, and the SNP is named g.348A>G.

[0014] The porcine ADIPOQ gene sequence is shown in Chromosome 13, NC_010455.5 (124633906..124646237). The region from 2000bp upstream to 700bp downstream of the transcription start site of the ADIPOQ gene is listed in the sequence table (as shown in SEQ ID NO.1), the first bold underlined nucleotide "A" is the transcription start site, the second bold underlined nucleotide "A" is the SNP site g.348A>G 348bp downstream of the transcription start site, and the site is located in the first intron region.

[0015] Furthermore, the substance is the following (b1) or (b2) or (b3):

[0016] (b1) PCR primers for amplifying a pig genomic DNA fragment containing the SNP site g.348A>G in the ADIPOQ gene;

[0017] (b2) a PCR reagent containing the PCR primers described in (b1);

[0018] (b2) A kit containing the PCR primers described in (b1) or the PCR reagents described in (b2).

[0019] Furthermore, the PCR primers include an upstream primer ADIPOQ-5F and a downstream primer ADIPOQ-5R:

[0020] ADIPOQ-5F: 5'-ACCTTCTCCTCATTTCCATC-3'(SEQ ID NO.3)

[0021] ADIPOQ-5R: 5'-TTCTCACGAACCCTACCC-3' (SEQ ID NO. 4).

[0022] Furthermore, in the above application, the SNP site g.348A>G is genotyped, and the back fat thickness of individuals with the genotype of GG is higher than the back fat thickness of individuals with the genotype of AG or AA.

[0023] In a second aspect, the present invention seeks to protect the use of the aforementioned PCR primers in the preparation of a product for detecting the polymorphism or genotype of the aforementioned SNP site g.348A>G.

[0024] In a third aspect, the present invention claims protection for a product for detecting the polymorphism or genotype of the above-mentioned SNP site g.348A>G, containing the above-mentioned substance or the above-mentioned PCR primer. The product may be a PCR reagent or a kit.

[0025] In a fourth aspect, the present invention claims protection for a method for identifying or assisting in identifying pig back fat thickness, detecting the polymorphism or genotype of the SNP site g.348A>G in the pig genome; the back fat thickness of individuals with the GG genotype is higher than the back fat thickness of individuals with the AG or AA genotype.

[0026] In the fifth aspect, the present invention seeks to protect a genetic breeding method for a pig variety with a high lean meat rate, detecting the polymorphism or genotype of the SNP site g.348A>G in the pig genome; retaining individuals with the AA homozygous genotype and the AG heterozygous genotype, and eliminating individuals with the GG homozygous genotype, thereby increasing the lean meat rate of pigs from generation to generation.

[0027] Furthermore, in the above method, the method for detecting the polymorphism or genotype of the SNP site g.348A>G in the ADIPOQ gene on the pig genome is: using the pig genome DNA to be tested as a template, using the aforementioned PCR primers or the aforementioned product to perform PCR amplification; using the PCR-RFLP-Ssil method to perform enzyme digestion typing on the amplified fragment, the enzyme digestion products of different genotypes will show different numbers of bands: the GG genotype has two bands of 363bp and 212bp, the AG genotype has three bands of 575bp, 363bp and 212bp, and the AA genotype has one band of 575bp.

[0028] In a specific embodiment of the present invention, the method for detecting the polymorphism or genotype of the SNP site g.348A>G in the ADIPOQ gene on the pig genome specifically comprises the following steps:

[0029] 1) Extracting DNA from the pig tissue sample to be tested;

[0030] 2) using the above-mentioned PCR primers or the above-mentioned products to perform PCR amplification on the DNA obtained in step 1) to obtain an amplified fragment;

[0031] 3) The amplified fragments were digested and typed by the PCR-RFLP-SsiI method. The digested products of different genotypes showed different numbers of bands: the GG genotype had two bands of 363 bp and 212 bp, the AG genotype had three bands of 575 bp, 363 bp and 212 bp, and the AA genotype had one band of 575 bp;

[0032] 4) When the typing result is GG type, it is judged that its back fat thickness is higher than the back fat thickness value of AG type or AA type individuals.

[0033] In a specific embodiment of the present invention, the PCR amplification reaction 25 μL system: Es Taq MasterMix (Dye) 22 μL, upstream and downstream primers 1 μL each, 1 μL of DNA to be detected; PCR reaction program: 94°C, 2 min; 94°C, 30 s; 54°C, 10 s annealing; 72°C, 30 s; 35 cycles; 72°C, 2 min.

[0034] The reaction system of the PCR-RFLP-SsiI method is: 8 μL of the PCR amplification product to be detected, 0.2 μL of Ssil enzyme, 10× FastDigest TM Buffer 1μL, ddHO 2 O 0.8μL; reaction conditions: 37℃, 10min.

[0035] In the sixth aspect, the present invention requests protection for a molecular marker containing the aforementioned SNP site g.348A>G, the nucleotide sequence of which is shown in SEQ ID NO.2, and the SNP site g.348A>G is located at the 348bp nucleotide downstream of the transcription start site of the ADIPOQ gene in the SEQ ID NO.2 sequence.

[0036] The room temperature described in the present invention is 25±10°C.

[0037] Beneficial effects of the present invention:

[0038] The present invention studies the correlation between the key gene ADIPOQ in lipid metabolism in organisms and the pig backfat thickness trait, screens out a SNP site related to the pig backfat thickness trait in the intron region, thereby obtaining functional genes and molecular genetic markers related to the pig backfat thickness, laying a foundation for further controlling the pig backfat thickness and assisting the genetic improvement of pigs to improve the lean meat rate of carcasses, so as to directly perform genotype selection or marker-assisted selection on pigs, thereby accelerating the selection and breeding of superior pig varieties. BRIEF DESCRIPTION OF THE DRAWINGS

[0039] Figure 1 It is the SNP site g.348A>G of the porcine ADIPOQ gene discovered by sequencing in the present invention.

[0040] Figure 2 The schematic diagram of the typing results of three genotypes (GG type, AG type and AA type) of the porcine ADIPOQ gene SNP site g.348A>G using the PCR-RFLP-Ssil method. Marker: 2000bp DNA molecular weight marker.

[0041] Figure 3 This is the back fat thickness of individuals with different genotypes at the SNP site g.348A>G of the ADIPOQ gene in a Du-Changda hybrid population. DETAILED DESCRIPTION

[0042] The present invention is further described below with reference to the examples. The experimental methods in the following examples without specifying specific conditions are generally carried out according to the well-known means in the art.

[0043] Example 1

[0044] 1. Experimental Materials

[0045] 1.1 Experimental animals

[0046] This study used 241 Duroc-Changda hybrid pigs. The experimental animals were from Jiangsu Changzhou Kangle Agriculture and Livestock Co., Ltd. and were raised under the same conditions. Liver tissue samples were collected for DNA extraction.

[0047] 1.2 Test reagents

[0048] (1) Conventional genomic DNA purification kit (containing RNase A): containing silica membrane centrifugal column, EB, BB2,

[0049] LB2, WB2, Proteinase K and RNase A were purchased from Beijing Quanshijin Biotechnology Co., Ltd. and stored at room temperature.

[0050] (2) 2×Es Taq Master Mix (Dye): Contains 2×Es Taq Master Mix (Dye) and ddH2O, purchased from Nanjing Qingke Biotechnology Co., Ltd. and stored at -20°C.

[0051] (3) Goldview nucleic acid gel dye: purchased from Hunan Aikerui Biotechnology Co., Ltd. and stored at 4°C.

[0052] (4) DL2000 Marker: purchased from Nanjing Qingke Biotechnology Co., Ltd., stored at -20°C, and stored at 4°C after thawing.

[0053] (5) Anhydrous ethanol AR 500mL: purchased from Shanghai Sinopharm Chemical Reagent Co., Ltd.

[0054] (6) FastDigest SsiⅠ restriction endonuclease: Contains FastDigest restriction endonuclease, 10×FastDigest TM Green Buffer and 10×FastDigest TM Buffer was purchased from Nanjing Thermo Fisher Scientific Co., Ltd. and stored at -20℃.

[0055] (7) 1×TAE buffer: Prepare by yourself. To prepare 50 mL of the system, you need 12.1 g of Tris base, 2.855 mL of glacial acetic acid, and 5 mL of EDTA (0.5 M, pH 8.0).

[0056] (8) Agarose powder: purchased from Nanjing Qingke Biological Co., Ltd.

[0057] 1.3 Instruments and Equipment

[0058] (1) CF1524R desktop high-speed micro-refrigerated centrifuge: purchased from Shanghai SCILOGEX Technology Co., Ltd.

[0059] (2) NanoDrop 2000 ultra-micro-volume spectrophotometer: purchased from ThermoFisher Technology Company

[0060] (3) PCR instrument: purchased from Zhuhai Dark Horse Medical Instrument Co., Ltd.

[0061] (4) Gradient PCR instrument: purchased from Bio-Rad Life Sciences Products, Inc., USA

[0062] (5) Horizontal electrophoresis apparatus: purchased from Beijing Junyi Oriental Electrophoresis Equipment Co., Ltd.

[0063] (6) Tanon 3500 gel imaging system: purchased from Shanghai Tianneng Technology Co., Ltd.

[0064] (7) Mini metal bath instrument: purchased from Zhejiang Qunan Laboratory Instrument (JOANLAB) Co., Ltd.

[0065] (8) Double-hole constant temperature water bath: purchased from Zhejiang Qunan Laboratory Instrument (JOANLAB) Co., Ltd.

[0066] The amplified fragments were sequenced by Qingke (Nanjing) Biological Company

[0067] 2. Experimental Methods

[0068] 2.1 Determination of carcass backfat thickness

[0069] After slaughtering the pigs, the back fat thickness was measured with a vernier caliper in cm. The measurement locations included the back fat thickness at the junction of the waist and sacrum, the thickest part of the shoulder, the 3rd and 4th ribs, and the average back fat thickness at three points.

[0070] 2.2 DNA extraction from liver tissue

[0071] Follow the steps in the instructions of the "Conventional Genomic DNA Purification Kit".

[0072] (1) Select liver tissue of Duroc-Changda hybrid pigs for sampling, mince the liver with scissors and forceps, control its weight to about 25 mg and put it into a 1.5 mL centrifuge tube. Add 100 μlL LB2 and 20 μL Proteinase K to completely immerse the tissue in the solution, preheat the constant temperature water bath (BHS-2, Qunan Testing Instrument Co., Ltd., Ningbo, Zhejiang) to 55°C and incubate for 3 hours to completely lyse the tissue sample. Centrifuge at 12,000×g for 5 minutes at 25°C, and transfer the supernatant to a new sterile centrifuge tube.

[0073] (2) Add 500 μL of BB2 to the supernatant, shake rapidly for 5 seconds to mix evenly, and incubate at room temperature for 10 min. Transfer all the liquid in the tube to a centrifuge column, centrifuge at 12,000 × g for 30 seconds at 25°C, and discard the flow-through.

[0074] (3) Add 500 μL of solution CB2 supplemented with anhydrous ethanol to the centrifuge column and continue centrifugation at 12,000 × g for 30 s at 25°C, then discard the flow-through.

[0075] (4) Then, add 500 μL of WB2 solution with anhydrous ethanol to the centrifuge column and centrifuge at 12,000 × g for 30 seconds at room temperature. Discard the effluent and repeat this step twice. To completely remove the WB2 remaining in the column, centrifuge again for 2 minutes at the same speed.

[0076] (5) After centrifugation, place the centrifuge column in a new clean centrifuge tube and add 50-200 μl of preheated solution EB (60℃~70℃) or deionized water (pH>7.0). Let stand at room temperature for 1 minute and then centrifuge at 12000×g for 1 minute. The liquid in the tube is the eluted DNA. If you want to get more DNA, repeat this step. The eluted DNA is stored at -20℃ for later use.

[0077] 2.3 DNA concentration determination and DNA sample pooling

[0078] NanoDrop2000 spectrophotometer was used to measure DNA concentration and purity, and the OD260 / OD280 ratio was calculated to determine the quality of DNA extraction. If the OD260 / OD280 ratio of all samples should be 1.8-2.0, it means that the quality of DNA extraction is qualified.

[0079] 2.4 Primer design and synthesis

[0080] According to the porcine gene sequence ADIPOQ (Chromosome 13, NC_010455.5 (124633906..124646237)) in the GenBank database, five pairs of primers were designed from 2000 bp before the transcription start codon of the promoter region to 700 bp downstream thereof, namely ADIPOQ-1F and ADIPOQ-1R, ADIPOQ-2F and ADIPOQ-2R, ADIPOQ-3F and ADIPOQ-3R, ADIPOQ-4F and ADIPOQ-4R, ADIPOQ-5F and ADIPOQ-5R. The primer sequences are shown in Table 1.

[0081] Table 1 Primer design information of porcine ADIPOQ gene

[0082]

[0083] 2.5 PCR-RFLP reaction system and procedure

[0084] The PCR reaction system used in this experiment is shown in Table 2 and Table 3:

[0085] Table 2 PCR reaction system

[0086]

[0087] Table 3 Genotyping restriction enzyme reaction system

[0088]

[0089]

[0090] PCR reaction procedure:

[0091] 94°C, 2 min;

[0092] 94°C, 30 s, 54°C, 30 s, 72°C, 30 s, 35 cycles;

[0093] 72℃, 2min; store at 4℃;

[0094] The amplified PCR product was digested with enzymes according to the system in Table 3. The reaction system was: 37°C, 10 min.

[0095] 2.6 Agarose gel electrophoresis detection

[0096] The amplified products and the digested products were electrophoresed with 2% agarose gel at a voltage of about 120V. After electrophoresis for about 30 minutes, the gel imaging system was used for observation. The PCR products obtained by successful amplification were digested. After the digestion reaction was completed, the digested products were subjected to agarose gel electrophoresis. Different genotypes corresponded to different numbers of bands. The genotype of the sample was determined based on the number of bands of each sample in the electrophoresis results.

[0097] 3. Experimental results

[0098] 3.1SNP screening

[0099] 20 DNA samples were randomly selected as templates, and the promoter region, 5′UTR region and intron region from 2000bp upstream to 700bp downstream of the transcription start site of the ADIPOQ gene were amplified using ADIPOQ-1F and ADIPOQ-1R, ADIPOQ-2F and ADIPOQ-2R, ADIPOQ-3F and ADIPOQ-3R, ADIPOQ-4F and ADIPOQ-4R, ADIPOQ-5F and ADIPOQ-5R, respectively, to obtain a total of about 2700bp fragments in the porcine ADIPOQ gene, and then the PCR products were sequenced (provided by Qingke Biotechnology (Nanjing) Technology Co., Ltd.). The sequencing results showed that only one SNP site was found in the fragment amplified by the primer pair ADIPOQ-5F and ADIPOQ-5R, which was located at 348bp downstream of the transcription start site of the ADIPOQ gene, belonging to the first intron region, and was adenine A or guanine G, respectively. Figure 1 As shown, the SNP was named g.348A>G.

[0100] 3.2 PCR-RFLP genotyping

[0101] For the above-mentioned SNP sites, primers ADIPOQ-5F and ADIPOQ-5R were used for PCR amplification, and the genotype was identified by digesting the PCR product with SsiⅠ restriction endonuclease. Through agarose gel electrophoresis, the digestion products of different genotypes will show different numbers of bands: the GG genotype has two bands of 363bp and 212bp, the AG genotype has three bands of 575bp, 363bp and 212bp, and the AA genotype has one band of 575bp. According to statistics, in the 241 Du Changda three-way hybrid population, there are 52 individuals with the GG genotype, 77 individuals with the AA genotype, and 112 individuals with the AG heterozygous type. The digestion results of some PCR products are shown as follows Figure 2 .

[0102] 3.3 Correlation between mutation sites and backfat thickness

[0103] The number of individuals with different genotypes was counted and the correlation between SNP and pig back fat thickness was analyzed. The results showed that there was a correlation between the two. The back fat of GG type individuals (0.9983±0.1941cm) was the highest, followed by AG type individuals (0.9253±0.1861cm), and the back fat of AA type individuals (0.8753±0.2173cm) was the lowest. The back fat thickness of GG type individuals was significantly higher than that of AG or AA genotype individuals (P<0.05). Figure 3 .

[0104] 3.4 Prediction of transcription factor binding sites

[0105] According to the prediction of the transcription factor binding site at the g.348A>G site of the ADIPOQ gene on the JASPAR database website, the prediction results are shown in Table 4 .

[0106] Table 4 Prediction results of transcription factor binding sites of g.348A>G in ADIPOQ gene

[0107] Base type Proposed binding transcription factor A SPI1 G ——

[0108] 4. Results Analysis

[0109] The mutation site screened by the present invention has a significant correlation with pig back fat thickness, can be developed as a SNP molecular marker and used to predict the back fat thickness of live pigs. For the ADIPOQ intron region g.348A>G, when the genotype is GG type, its back fat thickness is higher than that of individuals with AG or AA genotypes. When the g.348A>G site in the intron region of the ADIPOQ gene is A, the transcription factor SPI 1 may bind to it, thereby regulating the expression of the ADIPOQ gene, and further affecting the back fat thickness of the Du-Chang-Dou triple hybrid pig.

[0110] The present invention screened a SNP site related to pig backfat thickness in the ADIPOQ intron region, thereby obtaining a functional gene and molecular genetic marker related to pig backfat thickness. The molecular marker provided by the present invention can efficiently and accurately identify pig backfat thickness, and then judge the meat production performance of pigs, which has significant application value in the selection of breeding pigs and provides a reliable reference for the selection of high-quality pig breeds.

[0111] It can be known that the above embodiments are only exemplary implementations used to illustrate the principles of the invention, but the present invention is not limited thereto. Those skilled in the art can make various improvements and changes without departing from the essence of the present invention, and these improvements and changes also fall within the scope of protection of the present invention.

[0112] SEQ ID NO.1

[0113] The sequence is the region from 2000bp upstream to 700bp downstream of the transcription start site in the porcine ADIPOQ gene sequence Chromosome 13, NC_010455.5 (124633906..124646237). The yellow highlighted sequence is the region amplified by the ADIPOQ primers ADIPOQ-5F and ADIPOQ-5R; the first bold underlined nucleotide "A" is the transcription start site, and the second bold underlined nucleotide "A" is the SNP site g.348A>G 348bp downstream of the transcription start site, which is located in the first intron region.

[0114]

[0115]

[0116] SEQ ID NO.2

[0117]

Claims

1. Use of a substance for detecting the polymorphism or genotype of the SNP site g.348A>G in the ADIPOQ gene on the pig genome in any of the following: (a1) Application in identifying or assisting in identifying pig backfat thickness; (a2) Use in the preparation of a product for identifying or assisting in identifying pig backfat thickness; (a3) Application in selecting or assisting in selecting pig breeds with low backfat thickness; (a4) Use in the preparation of products for selecting or assisting in selecting pig breeds with low backfat thickness; (a5) Application in assisting the selection of high lean meat rate pig breeds; The SNP site g.348A>G is located at the 348 bp nucleotide downstream of the transcription start site in the SEQ ID NO.2 sequence of the pig ADIPOQ gene, and its polymorphism is A or G.

2. The use according to claim 1, characterized in that: The substance is (b1) or (b2) or (b3) as follows: (b1) PCR primers for amplifying a pig genomic DNA fragment containing the SNP site g.348A>G; (b2) a PCR reagent containing the PCR primers described in (b1); (b2) A kit containing the PCR primers described in (b1) or the PCR reagents described in (b2).

3. The use according to claim 2, characterized in that: The PCR primers include an upstream primer ADIPOQ-5F and a downstream primer ADIPOQ-5R: ADIPOQ-5F: 5'-ACCTTCTCCTCATTTCCATC-3' ADIPOQ-5R: 5'-TTCTCACGAACCCTACCC-3'.

4. The use according to claim 1, characterized in that: The SNP site g.348A>G was genotyped, and the back fat thickness of individuals with the GG genotype was higher than the back fat thickness of individuals with the AG or AA genotype.

5. Use of the PCR primers described in claim 2 or 3 in the preparation of a product for detecting the polymorphism or genotype of the SNP site g.348A>G described in claim 1.

6. A product, characterized in that The product contains the substance described in claim 2 or the PCR primer described in claim 3.

7. A method for identifying or assisting in identifying pig backfat thickness, characterized in that: Detect the polymorphism or genotype of the SNP site g.348A>G in the ADIPOQ gene on the pig genome as described in claim 1; the back fat thickness of individuals with the GG genotype is higher than the back fat thickness of individuals with the AG or AA genotype.

8. A genetic breeding method for a pig with a high lean meat rate, characterized in that: Detect the polymorphism or genotype of the SNP site g.348A>G in the ADIPOQ gene on the pig genome described in claim 1; retain individuals with AA homozygous genotype and AG heterozygous genotype, eliminate individuals with GG homozygous genotype, and improve the lean meat rate of pigs generation by generation.

9. The method according to claim 7 or 8, characterized in that: The method for detecting the polymorphism or genotype of the SNP site g.348A>G in the ADIPOQ gene on the pig genome is as follows: using the pig genome DNA to be tested as a template, and performing PCR amplification using the PCR primers described in claim 2 or 3 or the product described in claim 6; performing enzyme digestion typing on the amplified fragment by the PCR-RFLP-SsiI method, and the enzyme digestion products of different genotypes will present different numbers of bands: the GG genotype has two bands of 363bp and 212bp, the AG genotype has three bands of 575bp, 363bp and 212bp, and the AA genotype has one band of 575bp.

10. A molecular marker comprising the SNP site g.348A>G of claim 1, characterized in that: The nucleotide sequence of the SNP molecular marker is shown in SEQ ID NO.2, and the SNP site g.348A>G is located at the 348 bp nucleotide downstream of the transcription start site of the ADIPOQ gene in the SEQ ID NO.2 sequence.

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