金川牦牛BCL2A1基因克隆及生物信息学分析

为探讨金川牦牛抗凋亡因子--B淋巴细胞瘤2相关蛋白A1（B cell lymphoma 2 related A1,BCL2A1）基因特点,本试验采用PCR方法以金川牦牛血液DNA为模板扩增BCL2A1基因,用DNAStar、ExPASy、ABCpred等生物信息学软件分析基因序列和蛋白质结构。结果显示,克隆获得的片段长622 bp,GenBank登录号：MG459158,开放阅读框为516 bp,共编码171个氨基酸,其中缬氨酸（V）、赖氨酸（K）的含量较多,分别为9.4%和8.8%。BCL2A1分子质量为19.46 ku,理论等电点为4.99,不稳定系数为17.44,具有跨膜螺旋结构域（Scores>500）。二级结构主要为α-螺旋,占60.82%,有一个BCL2结构域,三级结构模型与人BCL2A1同源性最高（72.79%）,BCL2A1存在潜在的B细胞抗原表位。与野牦牛氨基酸序列进行比对显示,金川耗牛BCL2A1存在5个氨基酸差异位点。NJ法系统进化分析显示,金川牦牛与野牦牛同源性为98.5%,亲缘关系最近。本试验成功克隆了金川牦牛BCL2A1基因,其在哺乳动物中具有较高的保守性,为深入研究牦牛BCL2A1基因功能提供理论依据。

Cloning and Bioinformatics Analysis of BCL2A Gene in Jinchuan Yak

This study was aimed to explore the characteristics of B cell lymphoma 2 related A1 (BCL2A1) gene in Jinchuan yak.BCL2A1 gene was amplified by PCR using Jinchuan yak blood DNA as a template,and the gene sequences and protein structures were analyzed using DNAStar,ExPASy,ABCpred and other bioinformatics softwares.The results showed that the length of BCL2A1 gene was 622 bp,GenBank accession number:MG459158.The open reading frame was 516 bp,which could encode 171 amino acids with molecular weight of 19.46 ku.The higher proportions of amino acids were Val (V) and Lys (K),up to 9.4% and 8.8%,respectively.The theoretical isoelectric point was 4.99.The unstability index was 17.44,with transmembrane helices(Scores>500).The main secondary structure was alpha helix of 60.82%.There was a BCL2 domain,the tertiary structure model had the highest homology with human BCL2A1 (72.79%).BCL2A1 had a B cell potential epitope.Alignment with Bos mutus amino acid sequence showed that there were five amino acids mutations.Phylogenetic relationships revealed that the genetic relationship was the closest between Jinchuan yak and Bos mutus,and the homology was 98.5%.The BCL2A1 gene of Jinchuan yak was cloned successfully,which was highly conserved in mammals.It provided a theoretical evidence for further study on the function of BCL2A1 gene in yak.