Detection of two novel single nucleotide polymorphism (SNP) sites of promoter region of sheep myostatin (MSTN) gene and establishment of detection method thereof

Abstract

The invention discloses the detection of two novel single nucleotide polymorphism (SNP) sites of a promoter region of a sheep myostatin (MSTN) gene, and the establishment of a detection method thereof. The method comprises the following steps of: 1, determining two mononucleotide mutational sites of the promoter region of the sheep myostatin gene, wherein the two mononucleotide mutational sites are positioned in a GeneBank sequence, the registry number is DQ530260, and the SNP sites (-959T / C, -784G / A) are positioned at -959th site and -784th site according to an initial codon ATG and comprise 6 genotypes (TT, TC, CC, GG, GA, AA); 2, detecting three specific primers of the mutant allele, wherein the lengths of the three specific primers are 22bp, 22bp and 21bp respectively, the first primer is used for the sequencing of the promoter region of the MSTN gene, and the second and third primers are bonded onto template strands of two mutant alleles respectively and are amplified to target genes obtained by the mutation of the alleles; and 3, performing polymerase chain reaction (PCR) amplification on genomic deoxyribonucleic acid (DNA) of a sample by a designed oligonucleotide primer, performing enzyme digestion analysis on products which are amplified to 985bp and 121bp fragments by utilizing Psp1406I and Rsa I restriction enzymes, and establishing a PCR-RFLP detection system of the SNP sites so as to obtain the polymorphism of the two mononucleotide mutational sites.

Description

technical field [0001] The invention relates to two single nucleotide polymorphism sites in the promoter region of sheep myostatin (MSTN) gene and a detection method thereof. Background technique [0002] The growth traits of sheep muscle (including yield and quality) determine the production performance of meat sheep. The meat production performance is not only related to the nutrition and feeding management factors of sheep, but also mainly depends on genetic factors, that is, it is mainly regulated by genes related to muscle growth and development. In the past ten years, a large number of studies have shown that there are QTLs that affect sheep muscle growth traits on sheep chromosome 2. Myostatin gene (Myostatin, MSTN) is a member of the transforming growth factor superfamily. It is an important class of negative regulators of muscle growth discovered in recent years. The mutation inactivation of this gene can block the effect of MSTN on the proliferation and growth of ...

AI Technical Summary

Problems solved by technology

So far, no SNP sites related to muscle growth have been found in the MSTN coding region of Texel sheep with double muscle trait and other breeds of sheep

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