SNP molecular marker of swine and primer thereof

A technology of molecular markers and markers, applied in the field of genetic bioengineering

Inactive Publication Date: 2010-05-12
SHANGHAI ACAD OF AGRI SCI
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although the existing technology has carried out RFLP analysis to it, but RFLP is used for the identification of variety fingerprint, the detectio

Method used

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  • SNP molecular marker of swine and primer thereof
  • SNP molecular marker of swine and primer thereof
  • SNP molecular marker of swine and primer thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] 1. Search for molecular markers

[0030] (1) Construction of DNApool

[0031] Randomly collect the ear tissues of 2 large white pigs, Meishan pigs, and Duroc individuals (regardless of sex), and after extracting DNA, absorb the DNA of 6 individuals in equal amounts and put them into the same centrifuge tube, mix well and set aside;

[0032] (2) Primer design

[0033] Using human PSMB6 gene cDNA (GenBank accession number: NM_002798) as an information probe, using the BLAST tool in NCBI to perform homologous sequence screening in the GenBank pig EST database, a series of ESTs with a homology of more than 90% (the fragment length is greater than 100bp), and then use the ASSEMBLY program in DNAstar to construct pig EST-contigs. Amplification primers were designed according to the spliced ​​sequence of EST.

[0034] Table 1 Primers, annealing temperature and expected length for molecular marker search

[0035]

[0036]

[0037] The total volume of the PCR reaction ...

Embodiment 2

[0041] 2. Distribution of alleles

[0042] (1) Design of test groups:

[0043] Experimental group: collect Meishan pigs (33 heads), Tongcheng pigs (50 heads), Qingping pigs (38 heads), Tibetan pigs (28 heads), Min pigs (23 heads), Erhualian pigs (43 heads), Large White pigs (28), Landrace (15), Pietrain (12), Duroc (14), Shannon (31), Pishen (42), Dashen (34) and Duchene ( 23) individual ear tissues, DNA was extracted, a total of 414 DNA samples. The purpose of the test population is to detect the distribution of SNP markers in different varieties.

[0044] (2) Genotype detection: use the primer 1 detected in the previous step to detect the molecular markers, and use the same method to detect all individual genotypes of the test population.

[0045] (3) Statistical analysis: record the genotypes of all individuals, and calculate the allele frequency.

[0046] Table 2 The distribution of PCR-RFLP-Csp6I polymorphism in 13 pig breeds or strains

[0047]

[0048] Analyzing...

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Abstract

The invention screens a PSMB6 gene through swine breeds and combination widely used in commercial swine breeding, and searches out an SNP molecular marker for tracing DNA of a pork product; and a molecular marker of a (C/G) multiple allele SNP is in the 428th locus in the gene. The invention also provides a group of primers for identifying the SPN molecular marker of a swine PSMB6 gene sequence and a method thereof. The analysis of the distribution condition of the SNP locus allele can be used for tracing the DNA of the pork product.

Description

technical field [0001] The invention relates to the field of genetic bioengineering; further, a new SNP molecular marker is identified in pig breeds widely used in commercial pigs by using the pig PSMB6 gene, and the molecular marker can be used for DNA traceability of pork products. Background technique [0002] Food is the most important thing for the people. With the improvement of living standards and health care awareness, people pay more and more attention to food safety issues. As an important protein source in human diet, pork products have become an important part of food safety. Potential safety hazards of meat products are mainly excessive drug residues and zoonotic diseases. Therefore, the traceability management of meat products is realized, including breeding, transportation, slaughtering, segmentation, sales and other links. Through the traceability management of meat products, consumers can be provided with accurate and detailed information about the produc...

Claims

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Application Information

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IPC IPC(8): C12N15/12C12Q1/68C12N15/11
Inventor 唐雪明吴潇
Owner SHANGHAI ACAD OF AGRI SCI
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