威宁绵羊和贵州半细毛羊FecB基因SNP分析

SNP analysis of Fec B in Guizhou semi-fine wool sheep and Weining sheep

利用威宁绵羊和贵州半细毛羊构建DNA池,设计11对引物扩增其Fec B基因外显子和部分内含子的序列,对纯化后的PCR产物双向测序,分析确定多态性位点。然后利用生物信息学软件分析SNPs位点对Fec B基因RNA二级结构、Fec B蛋白结构的影响。结果显示,在扩增的Fec B基因中筛选到10个SNPs,其中exon5-A39G、exon9-C37A、exon11-C87A、intron2-A62G、intron4-A1023G、intron10-G24A突变为威宁绵羊和贵州半细毛羊中所共有,而exon9-A8G和intron1-G243A突变为威宁绵羊所特有,intron3-G177A和exon8-T86C突变为贵州半细毛羊所特有。上述的SNPs中,exon5-A39G和exon9-A8G为错义突变,exon5-A39G突变导致编码的异亮氨酸(Ile)变为缬氨酸(Val),exon9-A8G突变导致编码的天冬酰胺(Asn)变为天冬氨酸(Asp);exon9-C37A和exon11-C87A多态位点均未改变氨基酸的编码,为同义突变。

The DNA pond was constructed by using Weining sheep and Guizhou semi-fine wool sheep. 11 pairs of primers were designed to amplify exons and part of intron sequences of Fec B gene,then the purified PC R products were bi-directional sequenced to analyze polymorphic sites. Bioinformatic software was used to analyze the influences of SNPs sites on RNA secondary structure and protein structure. The results showed that 10 SNPs were screened,of which exon5-A39 G,exon9-C37 A,exon11-C87 A,intron2-A62 G,intron4-A1023 G,intron10-G24 A mutations were located in both Weining sheep and Guizhou semi-fine wool sheep,however,exon9-A8 G and intron1-G243 A mutations were peculiar to Weining sheep,and exon8-T86 C and intron3-G177 A mutations were peculiar to Guizhou semi-fine wool sheep. Exon5-A39 G and exon9-A8 G were missense mutations in the above SNPs. The exon5-A39 G mutation led to encoding isoleucine（Ile） into valine（Val） and the exon9- A8 G mutation led to encoding asparagine（Asn） into aspartic acid（Asp）. The exon9-C37 A and exon11-C87 A polymorphisms which were synonymous mutations did not change amino acid encoding.