Method for rapidly detecting single nucleotide polymorphism (SNP) of bovine PNPLA3 (patatin like phospholipase domain-containing 3) gene and application of method

Abstract

The invention discloses an RFLP (Restriction Fragment Length Polymorphism) method for rapidly detecting single nucleotide polymorphism (SNP) of bovine PNPLA3 (Patatin Like Phospholipase Domain-Containing 3) gene. The RFLP method comprises the steps of by taking PNPLA3 gene-containing to-be-detected whole-genome DNA as a template, and human-designed primer pairs P1-P5 as primers, carrying out PCR (Polymerase Chain Reaction) amplification on the bovine PNPLA3 gene, and discovering 27 SNP loci; selecting four missense mutation loci and respectively designing primer pairs P2-Bg1I, P2-RsaI, P3-BmgT120I and P3-MspI; carrying out PCR amplification by using the four primer pairs, respectively performing enzyme digestion to the PCR amplified product by using the four restriction endonuclease, and then detecting the enzyme digestion product by using agarose gel electrophoresis, wherein the electrophoresis result shows the SNP at the 2062th locus, the 2078th locus, the 35800th locus and the 35932th locus of the PNPLA3 gene of the Qinchuan bovine gene. The PNPLA3 gene relates to the growth traits of weight, average daily gain and the like, the PNPLA3 gene has the activity of lipase and acyltransferase and plays an important role in aspects of maintaining energy balance, the detection method can be used for marker assisted selection (MAS) breeding of the growth traits of China cattle, thus being beneficial to fast setting cattle population with excellent genetic resources.

Description

technical field [0001] The invention belongs to the field of animal molecular breeding, in particular to an RFLP (restriction fragment) for rapidly detecting the single nucleotide polymorphism (SNP) of the phospholipase-like phospholipase domain containing 3 (PNPLA3) gene length polymorphism) method, specifically, a restriction endonuclease is used to digest the gene sequence containing the single nucleotide polymorphism site, and the fragment size is determined according to agarose gel electrophoresis Separation, using the gel imaging system to analyze the size of the fragment, so as to determine the genotype of its SNP, because it has a significant correlation with Qinchuan's body weight and daily gain, it can be used as a molecular marker for molecular breeding of cattle. Background technique [0002] Single nucleotide polymorphism (SNP) refers to the polymorphism caused by the substitution of a single nucleotide (A / T / C / G) in the genomic DNA sequence. [0003] Cattle is ...

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Problems solved by technology

[0006] There is no research on the genetic variation of the bovine PNPLA3 gene at home and abroad, and the research on the function of this gene locus and the relationship between genetic variation and economic traits (such as body weight, etc.) are still blank

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