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SNP marker used for traceability in pig SFTPC 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 problems such as band mismatch and deletion, complex SSR marker band patterns, and slow template reaction performance

Inactive Publication Date: 2012-05-30
SHANGHAI ACAD OF AGRI SCI
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] AFLP markers are highly reliable and easy to operate, but at the same time, there are disadvantages such as slow response to templates, possible mismatch and deletion of bands, and relatively high cost.
Compared with SNP markers, SSR markers have many alleles and rich polymorphisms, but because of this, the band patterns of SSR markers are complicated, which brings difficulties to the automation and scale of DNA fingerprinting

Method used

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  • SNP marker used for traceability in pig SFTPC gene and detection method thereof

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] Molecular marker search

[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 cDNA sequence of the porcine SFTPC gene (Genbank: AJ891031) as a template, design primers to isolate the porcine SFTPC gene (using the DNA of a Duroc pig as a template for PCR amplification), the primers are as follows:

[0030] Forward primer: 5'-TACTTAGGAGGTCGCAGAA-3' (see sequence SEQ ID No 3)

[0031] Reverse primer: 5'-CCCGCTGGTAGTCATAGA-3' (see sequence SEQ ID No 4)

[0032] The total volume of the PCR reaction is 20μl, including about 100ng of porcine SFTPC genomic DNA, containing 1×buffer (Promega Company), 1.5mmol / L MgCl 2 , the final concentration of dNTP (Shanghai Sangon Biotechnology Co., Ltd....

Embodiment 2

[0045] Distribution of alleles

[0046] (1) Design of test groups

[0047] Experimental group: collected Pietrain (19 heads), Shannon (28 heads), Dashen (31 heads), Pishen (10 heads), Changshen (26 heads), Dushen (23 heads), Pi Dashen ( 18 individuals), Changchangshen (25 individuals), Du Dashen (7 individuals), Dupishen (8 individuals) and Dupi Dashen (31 individuals), and DNA was extracted from the ear tissue, a total of 226 DNA samples.

[0048] The purpose of the test population is to detect the distribution of SNP markers in different breeds.

[0049] (2) Genotype detection

[0050] Use forward primer: 5'-TACTTAGGAGGTCGCAGAA-3' (see SEQ ID No3)

[0051] Reverse primer: 5'-CCCGCTGGTAGTCATAGA-3' (see SEQ ID No4)

[0052] Amplification was performed and all individual genotypes of the test population were detected by the same RFLP method.

[0053] (3) Statistical analysis

[0054] The genotypes of all individuals were recorded, and the allele frequency and heterozygosi...

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Abstract

The invention relates to a SNP marker used for traceability in a pig SFTPC gene and a detection method thereof. The SNP molecular marker is obtained through DNA pool sequencing, and has a sequence as shown in SEQ ID NO 1; a base is mutated from 639A to 639T at the 639th base of the SNP molecular marker, and the coded amino acid sequence is shown in SEQ ID NO 2. In a test group including 11 pig varieties or strains, the allele distribution condition of the SNP molecular marker in the test group is analyzed, and the allele frequencies of the molecular marker in different varieties or strains are found to be approximate; the polymorphism is abundant; the allele frequency distribution difference between varieties or strains is fewer; the heterozygosity is more than 0.3; and the SNP marker is preliminary determined to be applicable to DNA traceablility for pork products.

Description

technical field [0001] The invention belongs to the field of food safety, and in particular relates to a traceable SNP molecular marker in pig SFTPC 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 unsaf...

Claims

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Application Information

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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蒋玲曦陶世如
Owner SHANGHAI ACAD OF AGRI SCI
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