猪A组轮状病毒VP7基因的克隆及生物信息学分析

Cloning and Bioinformatical Analysis of VP7 Gene in Porcine Group A Rotavirus

为了研究猪A组轮状病毒VP7基因功能,根据GenBank中猪轮状病毒VP7基因DNA序列设计引物,以实验室轮状病毒上海分离株SH-1为模板,进行PCR扩增,将VP7基因克隆到pMD-18T载体后,进行序列测定及分析。结果表明:此序列编码326个氨基酸,其相对分子量为37.38 Ku,理论等电点(PI)为4.77。VP7蛋白以α螺旋为主,主要有13个抗原表位区域。系统进化树分析显示分离株SH-1与KM230930.1株亲缘关系最近,基因型分析结果是G10型,属于人兽共患病毒株。

In order to investigate the function of porcine group A rotavirus VP7 gene,a set of primers were designed based on the DNA sequence of porcine rotavirus VP7 gene in GenBank.The rotavirus Shanghai isolate SH-1 was used as a template for PCR amplification,and the obtained VP7 gene was cloned into pMD-18T vector for sequencing and analysis.The results showed that the VP7 gene sequence encoded 326 amino acids with relative molecular weight of 37.38 Ku and theoretical isoelectric point(PI)of 4.77.The VP7 protein was dominated by alpha helices and contained 13 epitope regions.Phylogenetic tree analysis showed that the isolate SH-1 was genetically closest to KM230930.1 strain.The genotype analysis demonstrated that the isolate SH-1 was G10 type which belonged to the zoonotic virus strain.