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A traceable SNP molecular marker of porcine sncg gene and its detection method

A technology of molecular markers and genes, applied in the determination/testing of microorganisms, biochemical equipment and methods, DNA/RNA fragments, etc., can solve the problem of lack of SNP markers

Active Publication Date: 2015-11-11
SHANGHAI ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[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, 18, etc. 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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  • A traceable SNP molecular marker of porcine sncg gene and its detection method
  • A traceable SNP molecular marker of porcine sncg gene and its detection method
  • A traceable SNP molecular marker of porcine sncg gene and its detection method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] Example 1 Search for Molecular Markers

[0026] (1) Construction of DNA pool (pool)

[0027] The ear tissues of 2 Duroc pigs and 2 Meishan pigs (regardless of sex) were randomly collected, and DNA was extracted, and the DNA of 4 individuals was extracted in equal amounts and put into the same centrifuge tube, mixed well and set aside.

[0028] (2) Primer design

[0029] Using the DNA sequence of the porcine SNCG gene (Genbank: EF104639) as a template, design primers to isolate the porcine SNCG gene (using the DNA of a Duroc pig as a template for PCR amplification), the primers are as follows:

[0030] Forward primer: 5'-TTTGTGATGGTGCTCTGCGTTT-3' (SEQ ID NO.2)

[0031] Reverse primer: 5'-CCCTGCCTGAAGGTGGTTGT-3' (SEQ ID NO.3)

[0032] The total volume of the PCR reaction is 20μl, including about 100ng of porcine SNCG genomic DNA, containing 1×buffer (Promega Company), 1.5mmol / LMgCl 2 , the final concentration of dNTP (Shanghai Sangon Biological Company) was 150 μmol / L...

Embodiment 2

[0044] The distribution situation of embodiment 2 alleles

[0045] (1) Design of test groups

[0046] Experimental group: collected Pietrain (21 heads), Shannon (17 heads), Dabai (43 heads), Dashen (52 heads), Changshen (16 heads), Dushen (23 heads), Pi Dashen (11 heads) DNA was extracted from the ear tissues of individuals of the head), Dachang Shen (25 heads), Du Dashen (7 heads) and Dupi Dashen (20 heads), totaling 235 DNA samples.

[0047] The purpose of the test population is to detect the distribution of SNP markers in different breeds.

[0048] (2) Genetic testing

[0049] Forward primer: 5'-TTTGTGATGGTGCTCTGCGTTT-3' (SEQ ID NO.2)

[0050] Reverse primer: 5'-CCCTGCCTGAAGGTGGTTGT-3' (SEQ ID NO.3)

[0051] Perform amplification (conditions as in Example 1), and use the same PCR-Bpu1102I-RFLP method to detect all individual genotypes of the test population.

[0052] (3) Statistical analysis

[0053] The genotypes of all individuals were recorded, and the allele frequ...

Embodiment 3

[0057] Example 3 SNP molecular marker usability traceability test verification

[0058] The above-mentioned SNP molecular markers have been preliminarily confirmed to be used in traceability markers. In order to ensure the usability in traceability markers, the present invention re-samples at different locations and conducts usability traceability test verification.

[0059] Randomly collect 100 ear tissue samples and 100 muscle samples from individual pigs at Fuxing Slaughterhouse (every pig individual sample is collected at the same time as ear tissue samples and muscle samples), ear tissue samples are numbered E1-E100, and muscle samples are numbered M1-M100, respectively DNA from muscle samples and ear tissue samples was used for future use.

[0060] Detect the Bpu1102ISNP site of the pig SNCG gene of the present invention in 100 individual ear tissue samples and in 20 muscle samples randomly selected and the genotypes of the existing 11 SNP sites disclosed in the followin...

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Abstract

The invention discloses a SNP molecular marker in a porcine SNCG gene for tracing and a detection method thereof. The SNP molecular marker is obtained by DNA pool sequencing and has altogether 1599 bp. The SNP molecular marker contains a part of the sequence of a third exon; a base at a position 984 mutates from 984C to 984T; the DNA sequence of the SNP molecular marker is represented by SEQ ID NO. 1. The results of analysis of the allelic distribution of the SNP molecular marker in a trial group containing 11 pig species / strains show that allelic frequencies of the SNP molecular marker in different species or strains are approximate, polymorphism is rich, allelic frequency distribution difference between species or strains is small, the degree of heterozygosis is all larger than 0.3, and thus, the SNP molecular marker disclosed herein can be used for DNA tracing for pork products.

Description

technical field [0001] The invention belongs to the field of food safety, and relates to a traceable SNP molecular marker of a porcine SNCG 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 hidden ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/11C12Q1/68
Inventor 吴潇唐雪明蒋玮吕贝贝王金斌赵凯白蓝刘华武国干杨世方何建华
Owner SHANGHAI ACAD OF AGRI SCI
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