Method for rapidly detecting third exon single base mutation of myostatin gene

Abstract

The invention provides a method for rapidly detecting the third exon single nucleotide polymorphism (SNP) of a beef myostatin gene. In the method, aiming at the G-to-A mutation of a third exon of the myostatin gene, two groups of PCR primer pairs for respectively amplifying an upstream sequence and a downstream sequence of a G938A single base polymorphism site are designed, wherein the primer pairs are respectively provided with a mismatched primer for mutation site, and the 3' tail end of the primer is positioned on the mutation site. By utilizing the primers, different beef DNA samples can obtain different amplification results, thereby judging the polymorphism of the third exon of the myostatin gene. The method has low cost, and simple, rapid and accurate operation, is suitable for popularization and application, and is important to rapidly and accurately carry out marker-assisted selection of beef, accelerate the beef selecting and breeding process and research a rapid PCR detecting method of the field of molecular biology.

Description

technical field [0001] The invention relates to the field of molecular biology, in particular to a rapid detection method for detecting the single nucleotide polymorphism (SNP) of the third exon of bovine myostatin gene. Background technique [0002] Myostatin (myostatin), also known as growth differentiation factor 8 (GDF-8), is transforming growth factor β (transforming growth factor TGF-β) superfamily member, research results show that its function is a negative regulator of skeletal muscle growth and development, so the mutation of GDF-8 is the cause of bovine double muscle. The double-muscle individual of Belgian blue cattle has 11bp (937-947) deletion in exon 3, which causes a frameshift mutation, which leads to the stop of translation from the 14 codons after the deletion. As a result, only 7 amino acids at the C-terminal are translated, and the subsequent All 102 amino acids (274-375) are lost, so the translated protein cannot normally perform its biological functi...

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Problems solved by technology

[0011] Studies have shown that CRS-PCR is an effective method for detecting SNPs at known mutation sites. It is more flexible and can identify genotypes, which broadens the scope of application of the RFLP method; however, the choice of mismatched primers is small and difficult At the same time, the selection of mismatched bases should consider whether the corresponding restriction endonucleases are easy to purchase and the cost of the restriction endonucleases after the formation of restriction sites. It is difficult to distinguish genotypes, and the operation is cumbersome and time-consuming (Zhao Chunjiang et al., 2003; Zhang Zhongbin et al., 2004; Kwok S et al., 1990)

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