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SNP molecular maker in pig PSMA1 gene for traceability and detection method thereof

A molecular marker and detection method technology, applied in DNA/RNA fragments, recombinant DNA technology, etc., can solve the problems of band mismatch and deletion, complex band pattern and high cost

Inactive Publication Date: 2012-04-04
SHANGHAI ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] AFLP has high reliability and easy operation, but AFLP has the disadvantages of slow response to template, possible mismatch and deletion of bands, and relatively high cost.
Compared with SNP markers, microsatellite 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 molecular maker in pig PSMA1 gene for traceability and detection method thereof
  • SNP molecular maker in pig PSMA1 gene for traceability and detection method thereof
  • SNP molecular maker in pig PSMA1 gene for traceability and detection method thereof

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] Molecular marker search

[0028] (1) Construction of DNA pool (pool)

[0029] 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.

[0030] (2) Primer design

[0031] Using the cDNA sequence of the human PSMA1 gene (NM_148976) (http: / / www.ncbi.nlm.nih.gov / ) as a template, primers were designed to isolate the DNA fragment of the porcine PSMA1 gene (the DNA of a Duroc pig was used as PCR amplification template), the primers are as follows:

[0032] Forward primer: 5'-ctctgcctgtgtctcgtctt-3' (see sequence SEQ ID NO 5)

[0033] Reverse primer: 5'-gtacgagctgattgagaacg-3' (see sequence SEQ ID NO 6)

[0034] The total volume of the PCR reaction is 20μl, including about 100ng of porcine genomic DNA, containing 1×buffer (Promega Company), 1.5mmol / L MgCl 2 , the final concentration of dN...

Embodiment 2

[0052] Distribution of alleles

[0053] (1) Design of test groups

[0054] Experimental group: Erhualian pigs (23 heads), Meishan pigs (33 heads), Shanghai White pigs (11 heads), Shawutou pigs (30 heads), Large White pigs (28 heads), Landrace pigs (15 heads), Pietrain pigs (15 heads) were collected. (15 heads), Duroc (15 heads) and the cultivar Shannon (34) individuals ear tissues, DNA was extracted, a total of 204 DNA samples.

[0055] The purpose of the test population is to detect the distribution of SNP molecular markers in different varieties.

[0056] (2) Genotype detection

[0057] Use forward primer: 5'-ctctgcctgtgtctcgtctt-3' (see SEQ ID NO 5)

[0058] Reverse Primer - New: 5'-gcaccttatttagaacccac-3' (see SEQ ID NO 7)

[0059] Amplification was performed and all individual genotypes of the test population were detected by the same RFLP method.

[0060] (3) Statistical analysis

[0061] The genotypes of all individuals were recorded, and the allele frequency and ...

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Abstract

The invention relates to an SNP molecular maker in a pig PSMA1 gene for traceability and a detection method thereof. The SNP molecular maker is obtained by DNA pooling and sequencing and is represented by SEQ ID NO.3, and an amino acid sequence encoded by the SNP molecular maker is represented by SEQ ID NO.4. The SNP molecular maker is in a DNA fragment of the pig PSMA1 gene, wherein, the DNA fragment of the pig PSMA1 gene is represented by SEQ ID NO.1, and an amino acid sequence encoded by the DNA fragment of the pig PSMA1 gene is represented by SEQ ID No.2. The analysis of the allelic distribution of the SNP molecular maker in a group of trials containing 9 pig species / strains shows that allelic frequencies of the SNP molecular maker in different species or strains are approximate, the polymorphism is high, allelic frequency distribution difference between species or strains is small, and the heterozygosis degree is all larger than 0.3, thus the SNP molecular maker disclosed herein can be used for DNA technology for pork traceability.

Description

technical field [0001] The invention belongs to the field of food safety, and in particular relates to a traceable SNP molecular marker in porcine PSMA1 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 u...

Claims

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Application Information

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IPC IPC(8): C12N15/11C12Q1/68
Inventor 吴潇唐雪明张小波赵凯朱宏谭芙蓉蒋玲曦王金斌陶世如
Owner SHANGHAI ACAD OF AGRI SCI
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