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New molecular marker in MMP3 gene, used for DNA source tracing of pork products

A technology of molecular markers and genes, applied in the direction of DNA/RNA fragments, recombinant DNA technology, microbial measurement/inspection, etc., can solve the problems of label loss, blurred label pattern, easy label replacement, etc.

Inactive Publication Date: 2010-06-02
SHANGHAI ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In contrast, the label traceability technology adopted in my country has disadvantages such as label loss, record travel, ambiguous marking patterns, and easy artificial replacement of labels, while the DNA traceability technology is not affected by human factors, the results are accurate, and the detection methods are simple and fast, etc. Become the rapid traceability technology recognized internationally as the most development potential and application value

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  • New molecular marker in MMP3 gene, used for DNA source tracing of pork products

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0017] Molecular marker search

[0018] (1) Primer design

[0019] In the NCB database, using the mRNA sequence of the pig MMP3 gene (NM_001166308) as a template, Primer 5.0 was used to design amplification primers for finding new SNP sites.

[0020] Forward primer: 5`-GCAGAAGTTCCTTGGGTTGG-3`

[0021] Reverse primer: 5`-AAACTTTTCCAGGTCCGTCAA-3`

[0022] The total volume of the PCR reaction is 20μl, including about 100ng of porcine genomic DNA, containing 1×buffer (Promega), 1.5mmol / L MgCl 2 , the final concentration of dNTP is 150 μmol / L, the final concentration of primer is 0.2 μmol / L, 2U Taq DNA polymerase (Promega). The PCR amplification program was: 94°C for 4min, then cycled 35 times (94°C for 30s, annealing at 56°C for 30s, 72°C for 20s), and finally 72°C for 10min. PCR reaction products were detected by 1.0wt% agarose gel electrophoresis.

[0023] (2) Cloning and sequencing analysis: the obtained MMP3 gene PCR product is cloned according to the following method:

...

Embodiment 2

[0031] Distribution of alleles

[0032] (1) Design of test groups:

[0033] Experimental group: samples of Meishan pigs (33 heads), Ningxiang pigs (24 heads), Pudong White pigs (24 heads), Duroc (10 heads), American Large White (34 heads) and French Large White (65 heads) were collected. Ear tissue, DNA extraction, a total of 190 DNA samples. The purpose of the test group is to detect the distribution of SNP molecular markers in different varieties.

[0034] (2) Genotype detection: use forward primer and reverse primer to amplify and detect all individual genotypes of the test population by HpyF3I-RFLP.

[0035] (3) Statistical analysis: record the genotypes of all individuals, and calculate the allele frequency, see Table 1 for the results.

[0036] Table 1 Distribution of PCR-RFLP-HpyF3I polymorphism in 6 pig breeds or strains

[0037]

[0038]Analyzing the distribution of the alleles of the SNP site, it was found that the allele frequency of the molecular marker in d...

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Abstract

The invention discloses a new molecular marker in an MMP3 gene, used for DNA source tracing of pork products, which is acquired by the steps of analyzing distribution conditions of SNP locus alleles of the MMP3 gene of experiment groups (190 individuals in total) of 6 pig breeds or strains, clone sequencing as well as sequence analyzing and sequence comparing. The MMP3 gene sequence is 573 bp in length and contains sequences of a complete intron 2, an intron 3 and a complete exon 2 and sequences of part of exon 1 and part of exton 3; the MMP3 gene sequence is shown as SWQ ID NO 1, and a base mutation 182C-182T locates in a basic group NO.182 in the MMP3 gene sequence. Through a variation degree analysis of the alleles in the experiment groups, the SNP molecular marker is primarily predicated to be applied to the source tracing of the pork products.

Description

technical field [0001] The invention belongs to the field of food safety, and specifically relates to a new SNP molecular marker in the MMP3 gene used for DNA traceability of pork products. Background technique [0002] Fresh meat and meat products are indispensable food in people's daily life. Affected by our country's eating habits, pork is the most important meat product in people's daily life. Therefore, the safety of pork products has become the focus of meat product safety. Meat product traceability system, as a means of food quality and safety risk control and management, has been applied in more and more countries, and some countries even explicitly stipulate in the form of regulations that meat products that do not have traceability conditions are prohibited from entering the market. 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 hidden d...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12N15/11
Inventor 吴潇唐雪明赵凯朱宏谭芙蓉王金斌蒋玲曦陶世如
Owner SHANGHAI ACAD OF AGRI SCI