Authentication method of cattle CAPN1 gene as longisimus dorsi tenderness molecular marker and application

A CAPN1E14R, Longissimus muscle technology, applied in application, genetic engineering, plant genetic improvement, etc.

Inactive Publication Date: 2010-01-13
JILIN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

The experiment also found that the degradation bands produced by CAPN1 have certain differences

Method used

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  • Authentication method of cattle CAPN1 gene as longisimus dorsi tenderness molecular marker and application
  • Authentication method of cattle CAPN1 gene as longisimus dorsi tenderness molecular marker and application
  • Authentication method of cattle CAPN1 gene as longisimus dorsi tenderness molecular marker and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0088] Six individuals to be slaughtered were randomly selected from the beef cattle group of Haoyue Halal Meat Group Co., Ltd. in Changchun City, Jilin Province, China. Blood was collected during slaughter for genomic DNA extraction and the shear force of the longissimus dorsi muscle was measured. The extracted genomic DNA was determined by a spectrophotometer, the concentration was 36.54ug / ml, and the OD260 / OD280Ratio was 1.824. The primers, PCR reaction system and conditions designed by the present invention are used for PCR amplification. The PCR reaction system is: 10×PCR Buffer (purchased from Tiangen Biochemical Technology Co., Ltd.) 3.0 μl, dNTPs (purchased from Beijing Dingguo Biotechnology Company, the concentration is 2.5mM) 0.8 μl, forward primer and reverse primer (concentration 0.4 μl each of 10 pmol / μl), 0.4 μl of Taq enzyme (purchased from Tiangen Biochemical Technology Co., Ltd., at a concentration of 2.5 U / μl), 0.5 μl of genomic DNA (containing 18.27 ng DNA),...

Embodiment 2

[0102] Ten individuals to be slaughtered were randomly selected from the beef cattle group of Haoyue Halal Meat Industry Group Co., Ltd. in Changchun City, Jilin Province, China. Blood was collected during slaughter for genomic DNA extraction and the shear force of the longissimus dorsi muscle was measured. The extracted genomic DNA was determined by a spectrophotometer, the concentration was 34.54ug / ml, and the OD260 / OD280Ratio was 1.794. The primers, PCR reaction system and conditions designed by the present invention are used for PCR amplification. The PCR reaction system is: 10×PCR Buffer (purchased from Tiangen Biochemical Technology Co., Ltd.) 3.0 μl, dNTPs (purchased from Beijing Dingguo Biotechnology Company, the concentration is 2.5mM) 0.8 μl, forward primer and reverse primer (concentration 0.4 μl each of 10 pmol / μl), 0.4 μl of Taq enzyme (purchased from Tiangen Biochemical Technology Co., Ltd., at a concentration of 2.5 U / μl), 0.5 μl of genomic DNA (containing 17.27...

Embodiment 3

[0115] Nine individuals to be slaughtered were randomly selected from the beef cattle group of Haoyue Halal Meat Industry Group Co., Ltd. in Changchun City, Jilin Province, China. Blood was collected during slaughter for genomic DNA extraction and the shear force of the longissimus dorsi muscle was measured. The extracted genomic DNA was determined by a spectrophotometer, the concentration was 37.34ug / ml, and the OD26O / OD280 Ratio was 1.803. The primers, PCR reaction system and conditions designed by the present invention are used for PCR amplification. The PCR reaction system is: 10×PCR Buffer (purchased from Tiangen Biochemical Technology Co., Ltd.) 3.0 μl, dNTPs (purchased from Beijing Dingguo Biotechnology Company, the concentration is 2.5mM) 0.8 μl, forward primer and reverse primer (concentration 0.4 μl each of 10 pmol / μl), 0.4 μl of Taq enzyme (purchased from Tiangen Biochemical Technology Co., Ltd., at a concentration of 2.5 U / μl), 0.5 μl of genomic DNA (containing 18....

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Abstract

The invention belongs to the technical field of animal gene engineering, relating to clone of cattle longisimus dorsi tenderness related CAPN1 gene segment and an application thereof in cattle marker assisting selection. The obtained CAPN1 gene segment has the nucleotide sequence length of 520bp and comprises full fourteenth exons. A base mutation from A to G exists at the 166bp of the segment, which results in the generation of PsuI-RFLP restriction enzyme polymorphism. The mutation site is taken as a molecular marker, the restriction enzyme PsuI is used for detecting the mutation site, and association analysis can be carried out on the detection result and the cattle longisimus dorsi tenderness. Analysis result shows that striking difference exists among the longisimus dorsi tenderness of different genotype individuals, thus providing a theoretical basis for the seed selection and breeding of cattle. The invention is characterized by obtaining the CAPN1 gene part segment related to the cattle longisimus dorsi tenderness and providing the molecular marker and the application for the marker assisting selection of cattle by using a PCR-RFLP method to carry out polymorphism analysis on the segment.

Description

technical field [0001] The invention belongs to the technical field of animal genetic engineering, and in particular relates to the cloning of the CAPN1 gene fragment of the longissimus dorsi muscle tenderness-related gene fragment and its application in cattle marker-assisted selection. Background technique [0002] Experiments have shown (Wick M et al., J Biol Chem, 1999, 279: 1567-1582) that the degradation of major proteins in myofibrils is the fundamental reason for the improvement of meat tenderness, especially the weakening of the degradation of the Z disc, and the degradation of myofibrils. Degradation of connexin proteins (desmin and connexin) and intrasarcomatin proteins (titin, concatenin, and troponin-T) result in myofibril fragmentation, which constitutes the backbone of myofibrillar structural integrity. The degradation of these proteins is primarily responsible for tender maturation. It has also been proved by experiments (Koomaraie M, MeatSci, 1993, 32:93-10...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/57C12Q1/68
Inventor 姜昊张嘉保赵志辉刘颖高妍马腾壑秦立红戴立胜张金玉陈承祯
Owner JILIN UNIV
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