Separated nucleic acid sequence of total number of piglets related gene MMP3

A technology of total litter size and nucleic acid sequence, which is applied in the determination/inspection of microorganisms, DNA/RNA fragments, recombinant DNA technology, etc., can solve the problems of unsatisfactory breeding effect and low heritability, and achieve the goal of overcoming reproductive traits Insufficient effect of conventional selection

Inactive Publication Date: 2011-06-08
SHANGHAI JIAOTONG UNIV
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  • Summary
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AI Technical Summary

Problems solved by technology

Since reproductive traits such as total litter size of pigs are quantitative traits with low heritability, the breeding effect of conventional selection is not very ideal

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0019] Acquisition of Partial Genomic DNA Sequence of Porcine MMP3

[0020] (1) Primer design

[0021] Using the principle of homologous gene cloning, using human MMP3 gene eDNA (GenBank accession number: NM_002422) as a template, design a pair of primers with software Primer1.0:

[0022] PL: 5'-CTGGGCTATCAGAGGAAATG-3'

[0023] PR: 5'-CAGCATCCACTTTTGGTTCA-3'

[0024] (2) PCR reaction

[0025] 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 30 cycles of 94°C for 30s, 56°C for 30s, 72°C for 30s, and finally 72°C for 10min. PCR reaction products were detected by 1.5% agarose gel electrophoresis.

[0026] (3) Sequencing

[0027] After recovering the PCR product, connect it to PMD18-T Vector (TaKaRa). ...

Embodiment 2

[0033] Searching for Mutation Sites in Genome Sequence of Porcine MMP3

[0034] By sequencing the PCR amplification products of individual DNA from different pig breeds, and using biological analysis software to compare the sequences amplified by the same primer, it was found that the 816th position of the sequence shown in SEQ ID No: 1 of the MMP3 gene has There are two alleles of C and T, two alleles of T and G at the 919th position, and two alleles of A and T at the 1038th position.

Embodiment 3

[0036] Detection method of porcine MMP3 mutation site

[0037] (1) primer sequence (with embodiment 1)

[0038] PL: 5'-CTGGGCTATCAGAGGAAATG-3' (SEQ ID No: 2)

[0039] PR: 5'-CAGCATCCACTTTTGGTTCA-3' (SEQ ID No: 3 in the sequence listing)

[0040] (2) PCR amplification conditions

[0041] 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 35 cycles of 94°C for 30s, 56°C for 30s, 72°C for 30s, and finally 72°C for 10min. PCR reaction products were detected by 1.5% agarose gel electrophoresis.

[0042] (3) RFLP detection conditions

[0043] The volume of PCR product digestion reaction is 10μl, including 1μl of 1×buffer, 3~5μl of PCR product, 0.5μl (5U) of restriction endonuclease NheI or EcoRV, and use H ...

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Abstract

The invention relates to a separated nucleic acid sequence of a total number born trait related gene MMP3 of a pig in the technical field of genetic engineering. The separated nucleic acid sequence is provided with a sequence shown by an SEQ ID No:1, the length of which is 1346bp; the nucleic acid sequence shows a pair of allele-specific nucleic acid primers the lengths of which are 20bp; the pair of nucleic acid primers is specifically hybridized; and the sequence shown by the SEQ ID No:1 containing an eighth exon, a ninth exon and an eighth intron of the MMP3 gene of the pig is amplified. The implementation and the application of the separated nucleic acid sequence can be used for detecting and judging the quality of total number born traits of the pig; the molecular marker is applied to conduct a marker assisted selection during breeding of the pig, thereby correcting the defect during conventional selection of reproductive traits of the pig; and the separated nucleic acid sequencehas important practical significance.

Description

technical field [0001] The present invention relates to a nucleic acid sequence of pigs in the technical field of genetic engineering, in particular to an isolated nucleic acid sequence of a gene MMP3 related to pig total litter size traits. Background technique [0002] The total litter size of sows is an important economic trait, and increasing the total litter size of sows can bring huge economic benefits to modern pig production. Because reproductive traits such as total litter size of pigs are quantitative traits with low heritability, the breeding effect of conventional selection is not very ideal. With the in-depth study of pig molecular genetic markers and the application of some molecular experimental techniques, the differences between the genotypes of some main genes controlling quantitative traits are clearly displayed in front of the researchers, providing a basis for selection, Matching opens up new avenues. In 1994, Rothschild et al. found that the ESR gene ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/11C12Q1/68
Inventor 潘玉春吴潇高运臻谭桂芳王起山
Owner SHANGHAI JIAOTONG UNIV
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