贵州地方山羊ADAMTS1基因序列多态性及生物信息学研究

The Study of Sequence Polymorphism and Bioinformatics of Guizhou Native Goat ADAMTS1 Gene

本研究选择黔北麻羊和贵州黑山羊为试验对象,分别构建DNA池,结合PCR产物直接测序法分析两品种中ADAMTS1基因外显子2~8中可能与繁殖性状相关的单核苷酸多态位点。结果表明在黔北麻羊和贵州黑山羊两个群体中共检测到ADAMTS1基因的10个SNPs位点,其中该基因第2、5、6外显子中各存在1个SNP位点,依次是C266T、T2210C、C2513A,均引起所编码氨基酸的同义突变,第3、4、6、8内含子中各存在1个SNPs位点,依次是C1073T、A1326G、G2698A、T4261A,第7内含子中同时存在3个SNPs位点,为T3401C、T3045T、T3064C,其中9个SNPs位点(C266T,C2513A,C1073T,A1326G,G2698A,T4261A,T3401C,T3045G,T3064C)为两品种所共有,T2210C仅存在黔北麻羊中。位点A1326G的A和G等位基因频率在检测的两个羊种间差异较大,通过后续的生物信息学分析显示,ADAMTS1基因外显子2的C266T和外显子6的优势突变C2513A的位点变异共同引起m RNA二级结构发生显著改变。

Qianbei Ma goat an d Guizhou Black goat were selected as experimental subject, constructing DNA pool and combining direct sequencing technology of PCR product to analyse the single nucleotide polymorphism of A disintegrin and metallo-proteinase with thrombospondin motifs-1 gene exon 2 to exon 8 which was related possibly to reproduction traits in above two breeds. The result showed ten SNPs sites of ADAMTS1 gene were detected in both groups, among there was one SNPs found in the second, fifth, sixth exon respectively, they were C266 T, T2210 C, C2513 A resulting in coding amino acid's synonymous mutation, three SNPS sites-T3401 C,T3045T, T3064C- were found in intron 3 at the same time, nine SNPS(C266T, C2513 A, C1073 T, A1326 G, G2698 A,T4261A, T3401 C, T3045 G, T3064C) existed in commonly two breeds except for T2210 C only found in Qianbei Ma goat. Allele frequency of A1326 G site had much difference in different breeds, furthermore, bioinformatics analysis revealed the C266 T of exon 2 and dominant mutation C2513 A of exon 6 lead to mutualy the dramatical alteration of secondary structure of m RNA.