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Clone of pig intramuscular fat content gene Lpinl and uses thereof

A technology of intramuscular fat and genes, applied in the fields of application, genetic engineering, plant genetic improvement, etc., can solve complex physiological and biochemical processes and other problems

Inactive Publication Date: 2008-04-09
山东乾和源农牧科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although some important progress has been made in the research of candidate genes of pig intramuscular fat content, and several candidate genes have been revealed, the physiological and biochemical processes involved in pig intramuscular fat content are quite complex and may be regulated by multiple genes. Other novel genes with large effects remain to be discovered

Method used

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  • Clone of pig intramuscular fat content gene Lpinl and uses thereof
  • Clone of pig intramuscular fat content gene Lpinl and uses thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0025] Embodiment 1: the cloning of Lpin1 gene:

[0026] (1) Primer design: use human Lpin1 gene cDNA (GenBank accession number: NM_145693) as an information probe, use the BLAST tool in NCBI to screen homologous sequences in the Nucleotide database pig EST database, and obtain a series of similarities of more than 80% ESTs (fragment length greater than 100bp), use ENTREZ (http: / / www.ncbi.nlm.nih.gov / Web / Search / index.html) to query the corresponding sequence in NCBI with the accession numbers of these ESTs, and then use DNAStat Porcine EST-contigs were constructed using the SeqMan program in . The primers listed in Table 1 were designed according to the EST splicing sequence.

[0027] (2) Establish a 3′ RACE method: use the total RNA of adipose tissue as a template, use the primer anchor (see Table 1) to carry out reverse transcription to complete the synthesis of the first strand of cDNA, and then perform two rounds of PCR: using cDNA as a template, PCR amplification was ca...

Embodiment 2

[0041] Example 2: The application of the association analysis between Lpin1 gene C61-T61TaqI-RFLP genotype and some carcass traits and meat quality traits Establishment of PCR-RFLP diagnostic method

[0042] (1) PCR amplification conditions: use P-S1F and P-S1R as primers for PCR amplification, the total volume of 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 0.4 μmol / L, 2U Taq DNA polymerase (Promega). The PCR amplification program was: 94°C for 3min, 35 cycles of 94°C for 30s, 58°C for 30s, then 72°C for 25s, and finally 72°C for 5min. PCR reaction products were detected by 2% agarose gel electrophoresis.

[0043] (2) RFLP detection conditions: PCR product digestion reaction volume is 10μl, including 1×buffer 1μl, PCR product 4μl, restriction endonuclease TaqI 0.3μl (10U / μl), use H 2 Make up 10 μl of O, mix the sample a...

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Abstract

The invention pertains to the technical field of animal genetic engineering, in particular to an application of the clone of a fat content gene Lpin1 in the muscle of the swine and an application of using the gene Lpin1 as a molecular mark. The invention is characterized in that: parts of a cDNA sequence and a DNA sequence of the fat content gene Lpin1 in the muscle of swine are cloned as listed in the sequence tables of SEQID NO:1 and SEQID NO:3. Through the application of management analysis, the C61-T61 in table SEQID NO:3 is found to have obvious relation with the fat content in the muscle. The invention discloses the Lpin1 gene sequence of the swine, the gene mutation loci influencing the fat content in the muscle and a preparation method to obtain the gene and an application of the gene. The invention provides a new molecular mark for the marker-assisted breeding of the swine.

Description

technical field [0001] The invention belongs to the technical field of animal genetic engineering, and in particular relates to the cloning of a pig intramuscular fat content gene Lpin1 and its application as a molecular marker of the pig intramuscular fat content. Background technique [0002] Along with improving day by day of people's living standard, pork quality is more and more subject to people's attention. Although some highly cultivated lean meat varieties and strains in foreign countries have high feed conversion rate, fast growth and high lean meat yield, their meat quality is not good, and the probability of PSE meat is high, which often causes huge economic losses to the pig industry. Meat quality improvement has become the focus of current pig breeding work. The content of intramuscular fat (IMF) is a very important indicator for evaluating meat quality. IMF exists in the muscle, mainly on the epimysium, perimysium and endomysium. According to research, IMF c...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/12C12Q1/68
Inventor 刘榜何小平李奎赵书红樊斌余梅朱猛进彭中镇李长春徐学文
Owner 山东乾和源农牧科技有限公司
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