牦牛CAPN3基因CDS区克隆及生物信息学分析

Cloning and bioinformatics analysis on CDS of CAPN3 gene in yak

根据GenBank中普通牛CAPN3基因cDNA序列,设计特异性引物,采用RT-PCR技术从牦牛总RNA中克隆获得CAPN3基因完整的CDS区及部分3′UTR序列,预测分析该基因编码蛋白的生物学信息.结果表明:牦牛CAPN3基因CDS区全长2 469bp(GenBank登入号:JX112349.1),编码822个氨基酸残基(GenBank登入号:AFP73395.1);同普通牛比对,牦牛CAPN3基因在CDS区存在10个SNPs,并导致该蛋白功能区域内3个氨基酸突变;牦牛CAPN3基因编码蛋白分子量94.58ku,理论等电点(PI)5.70,其氨基酸序列中存在较强的亲水区域,二级结构为以α-螺旋和无规卷曲为主的混合型;牦牛CAPN3基因编码氨基酸序列与普通牛、绵羊、野猪等物种间同源性较高,系统进化情况与其亲缘关系远近一致.

The CDS region and partial sequence of 3＇UTR of the yak CAPN3 gene were cloned using RT-PCR with specific primers that designed according to cDNA sequence of bovine CAPN3 gene in the GenBank so as to analyze the bioinformatics of putative protein. The results showed that the yak CAPN3 gene contained a 2 469 bp CDS region （GenBank accession no.JX112349.1） and encoded a putative 822 a- mino acids （GenBank accession no：AFP73395.1）. Ten SNPs and corresponding three amino acid mutations in function region of protein were identified through alignment with bovine CAPN3 gene sequence . The putative molecular weight and theory isoelectric point of CAPN3 gene in yak were 94. 58 ku and 5.70, re- spectively. There was a hydrophobic region in ]]I region of yak CAPN3. The putative secondary structure of CAPN3 was mixed type with mainly s-helices and random coil. Deduced amino acid sequences of CAPN3 gene between yak and bovine, sheep,wild boar and other species were high on homology and the phylogenetic distance consistent with their genetic relationship.