威宁绵羊GHR基因多态性分析

The Polymorphism Detection of GHR Gene in Weining Sheep

为了更好地保护和开发利用威宁绵羊这一优良地方品种,本试验利用威宁绵羊构建DNA池,设计8对引物扩增威宁绵羊GHR基因外显子及部分内含子序列。将PCR产物纯化并进行双向测序,通过DNAStar和BLAST等生物软件分析确定SNP位点。利用生物信息学软件分析SNPs对GHR基因的m RNA二级结构的影响、生长激素受体二级和三级结构的改变。结果表明,在扩增的GHR基因中筛选到4个SNPs:Exon1-C^(112)T、Exon4-C^7T、Exon8-A^(27)T、Intron4-C^(27)T,其中Exon8-A^(27)T为错义突变,导致编码的异亮氨酸(Ile)变为苯丙氨酸(Phe);Exon1-C^(112)T和Exon4-C^7T位点对其编码的氨基酸没有影响,是同义突变;Intron4-C^(27)T在内含子区,不参与氨基酸编码。本研究将为更好地保护和开发利用威宁绵羊提供一些参考。

In order to protect and develop Weining sheep, we constructed DNA pooling and designed 8 primers to amplify Weining sheep GHR gene exon and intron sequence in this study. Then by using the technology of bi- directional sequencing, we determined the SNPs and bioinformatics software to analysis the influence of RNA secondary structure the secondary and tertiary structure of GHR gene. The result showed that there were 4 SNPs： Exonl-Cl12T, Exon4-C7T, Exon8-A27T （Ile-Phe）, Intron4-C27T （nonparticipation encoding amino acids）. This study might provide some references to protect and develop Weining Sheep.