利用HRM技术对皮南牛MSTN基因C313Y位点快速分型

旨在利用高分辨率熔解(HRM)技术建立一种皮南牛MSTN基因C313Y突变的快速分型方法,进而统计MSTN基因C313Y突变在皮南牛群体中的突变频率,以及突变对生产性能的影响。测量并统计99头皮南牛(母牛)的体尺和体质量指标,然后分别利用Sanger测序法和HRM技术对皮南牛MSTN基因C313Y位点进行分型,并利用一般线性模型分析突变对体尺和体质量指标的影响。结果表明,皮南牛在3岁之后体尺和体质量指标均趋于稳定,平均体高、体斜长、胸围、尻宽、尻长和体质量分别是(127.88±3.65) cm、(154.10±6.89) cm、(181.48±8.54)cm、(46.19±2.94) cm、(50.52±2.65) cm和(471.34±53.78)kg。Sanger测序法和HRM技术对MSTN基因C313Y位点的分型结果完全一致,其中野生型(GG)、杂合型突变(GA)和纯合型突变(AA)的个体分别占8.08%、70.7%和21.21%。关联分析表明,杂合型突变(GA)和纯合型突变(AA)均能够显著提高皮南牛的体尺和体质量指标,其中纯合突变型个体和杂合突变型个体的体质量比野生型个体分别显著提高38.55%和10.32%。成功建立基于HRM的皮南牛MSTN基因C313Y位点快速分型方法,该方法对于快速组建皮南牛MSTN C313Y纯合突变群体,进而有效提高群体的体尺和体质量指标具有重要的应用价值。

Rapid Genotyping of the MSTN C313Y in Pinan Cattle Using High-resolution Melting Technology

The aim of this study was to establish a rapid detection method for the MSTN C313Y mutation of Pinan cattle using high-resolution melting (HRM) technology, and then to analyze the mutation frequency and mutation effect in Pinan cattle. The body size and body mass of 99 Pinan cattle were measured, then the Sanger sequencing and HRM method were used to genotype the MSTN C313Y in experimental population, and the associations between the mutation and growth performance were analyzed by General Linear Model. The results showed that the body size and body mass of Pinan cattle became stable after the age of three. The average body height, body length, heart girth, rump width, rump length and body mass were (127.88±3.65) cm, (154.10±6.89) cm, (181.48±8.54) cm, (46.19±2.94) cm, (50.52±2.65) cm and (471.34±53.78) kg, respectively. The genotyping results of the Sanger sequencing and HRM method were identical, in which the wild type (GG), heterozygous mutant (GA) and homozygous mutant (AA) individuals accounted for 8.08%, 70.7% and 21.21%, respectively. Association analysis showed that both heterozygous mutant (GA) and homozygous mutant (AA) could significantly improve the body size and body mass of Pinan cattle, and the homozygous mutation had a greater improvement than the heterozygous mutation. Compared with wild type individuals, the body mass of individuals with homozygous mutant and heterozygous mutant increased by 38.55% and 10.32%, respectively. Our work demonstrates the successful establishment of HRM-based rapid genotyping method for the MSTN C313Y in Pinan cattle. This method has great application potentials for the rapid establishment of Pinan cattle core group with homozygous mutants, which can also effectively improve the body size and body mass of the whole group.