贵州白山羊和黔北麻羊TFAM 基因部分外显子多态性研究

利用黔北麻羊和贵州白山羊构建品种DNA池,设计1对引物分别扩增其TFAM基因第2外显子及第1内含子部分序列.PCR产物纯化后进行双向测序,DNAStar和BLAST分析确定多态性位点.利用生物信息学软件分析SNPs位点对TFAM基因RNA二级结构和TFAM蛋白二级、三级结构的影响.结果表明:在羊TFAM基因中筛选到5个SNPs:T 1005C、G1099C、A1130C、G1164C、T1287C,其中T1005C为同义突变,G1099C为错义突变,导致编码的甘氨酸(Gly)变为半胱氨酸(Cys);A1130C、G1164C、T1287C 3个突变位点均位于内含子区.SNPs位点对TFAM基因RNA二级结构和TFAM蛋白结构有一定影响.

Study on the SNP in part of TFAM exon of Whit Goat of Guizhou and Qianbei Ma Goat

Qianbei Ma Goat and White Goat of Guizhou were used to construct DNA pools, the complete sequences of extro 2 and part of intron 1 region in TFAM of the two breeds were obtained by designing one pair of specific primers. Then by conducting bidirectional sequencing after purification of PCR production subsequently. DNAStar and BLAST software were used to identify polymorphisms of goat TFAM genes. Moreover, through bioinformatics software to analyze the impact of secondary structure of RNA, secondary structure and tertiary structure of TFAM protein. The results showed that five SNPs (T1005C, G1099C, A1130C, G1164C, T1287C) were found in TFAM which included one missense mutation (G1099C, Gly--Cys), one synonymous mutation (T1005C, Val) and the other SNPs located in intron region respectively. The polymorphisms might lead to secondary structure change of RNA and the structure change of TFAM protein.