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Traceablility SNP marker in pig GIGYF2 gene and detection method thereof

A detection method and gene technology, applied in the direction of recombinant DNA technology, DNA/RNA fragments, etc., can solve the problems of small differences in allele distribution frequency, lack of SNP markers, and high degree of variability

Inactive Publication Date: 2012-06-06
SHANGHAI ACAD OF AGRI SCI
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0006] However, the SNP markers used for traceability of meat products are different from common SNP markers. For a single SNP marker, it must have at least the following characteristics: (1) High variability, close to allele frequencies in varieties or lines; (2) There is little difference in the distribution frequency of alleles among varieties; (3) the heterozygosity is greater than or equal to 0.3
[0007] In the long-term breeding work, researchers have accumulated a large number of SNP markers, but these SNP markers are often concentrated on certain chromosomes, such as chromosome 1, chromosome 12, etc., while other chromosomes, such as chromosome 5 and chromosome 8 Chromosomes 14 and 15 lack SNP markers, so in order to meet the requirements for DNA traceability of pork products, we conduct research on new SNP markers

Method used

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  • Traceablility SNP marker in pig GIGYF2 gene and detection method thereof
  • Traceablility SNP marker in pig GIGYF2 gene and detection method thereof
  • Traceablility SNP marker in pig GIGYF2 gene and detection method thereof

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Experimental program
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Effect test

Embodiment 1

[0023] SNP marker search

[0024] (1) Construction of DNA pool (pool)

[0025] The ear tissues of 2 Duroc pigs and 2 Shennon pigs (regardless of sex) were randomly collected respectively. After DNA was extracted, the DNA of 4 individuals was aspirated and put into the same centrifuge tube, and mixed well for use.

[0026] (2) Primer design

[0027] Using the cDNA sequence of the pig GIGYF2 gene (Genbank: NW 001885690) as a template, design primers to isolate the pig GIGYF2 gene (using the DNA of a Shannon pig as a template for PCR amplification), the primers are as follows:

[0028] Forward primer: 5'-TTTCTCCCTAAGTCGTCG-3' (see sequence SEQ ID No 2)

[0029] Reverse primer: 5'-CGTGCTGCTAAGTTTCTA-3' (see sequence SEQ ID No 3)

[0030] The total volume of the PCR reaction is 20 μl, including about 100 ng of pig GIGYF2 genomic DNA, containing 1 × buffer (Promega Company), 1.5 mmol / L MgCl2, dNTP (Shanghai Sangon Biological Company) final concentration of 150 μmol / L, and final p...

Embodiment 2

[0040] Distribution of alleles

[0041] (1) Design of test groups

[0042] Experimental group: collected Pietrain (24 heads), Shannon (32 heads), Dashen (31 heads), Pishen (10 heads), Changshen (26 heads), Pi Dashen (19 heads), Changchang Shen (25 heads), Du Dashen (7 heads), Du Pishen (8 heads) and Du Pi Dashen (31 heads) individual ear tissues, DNA was extracted, a total of 213 DNA samples.

[0043]The purpose of the test population is to detect the distribution of SNP markers in different breeds.

[0044] (2) Genotype detection

[0045] Use forward primer: 5'-TTTCTCCCTAAGTCGTCG-3' (see sequence SEQ ID No 2)

[0046] Reverse primer: 5'-CGTGCTGCTAAGTTTCTA-3' (see sequence SEQ ID No 3)

[0047] Amplification was performed and all individual genotypes of the test population were detected by the same RFLP method.

[0048] (3) Statistical analysis

[0049] The genotypes of all individuals were recorded, and the allele frequency and heterozygosity were calculated. The result...

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Abstract

The invention relates to a traceablility SNP marker in a pig GIGYF2 gene and a detection method thereof. The SNP marker which is positioned in a genomic DNA fragment in the pig GIGYF2 gene is obtained through DNA pool sequencing. The genomic DNA fragment in the pig GIGYF2 gene, which is represented by SEQ ID NO.1 and comprises bps with the number of 452, comprises parts of twenty-three introns, and there is a base mutation from 187A to 187C at the 187th bit. The distribution case of the SNP marker of the invention in allelic genes in a test colony comprising ten pig kinds or lines is analyzed, and results show that frequencies of the marker in the allelic genes of different kinds or lines are close, so the polymorphism is abundant, there is a small difference among the frequencies of the allelic genes of different kinds or lines, and heterozygosities are greater than 0.3, so a case that the SNP marker of the invention can be used for pig product DNA traceablility is preliminarily determined.

Description

technical field [0001] The invention belongs to the field of food safety, and in particular relates to a traceable SNP marker in the pig GIGYF2 gene and a detection method thereof. Background technique [0002] With the improvement of living standards and health awareness, people pay more and more attention to food safety issues. As an important source of protein in human diet, pork products have become a global hot issue on how to ensure their safety and sanitation and protect the health and life safety of consumers. Governments around the world have adopted a series of technical measures to strengthen the safety management of meat products in order to minimize the occurrence of food safety incidents. Major cities in my country have also established traceability systems for meat products. Through the traceability management of meat products, consumers can be provided with accurate and detailed information about products, which is conducive to timely discovery of unsafe hid...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/11C12Q1/68
Inventor 唐雪明吴潇张小波赵凯朱宏谭芙蓉王金斌魏曼曼陈凯
Owner SHANGHAI ACAD OF AGRI SCI
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