猪细小病毒HN-2011株VP2基因的克隆及序列分析

Cloning and Sequence Analysis of the VP2 Gene of Porcine Parvovirus HN-2011 Strain

根据GenBank中登录的猪细小病毒(PPV)VP2基因序列设计引物,扩增HN-2011河南分离株的VP2基因片段,并将其克隆到pMD 18T载体中。序列分析结果表明,HN-2011分离株VP2基因长度为1 740bp,编码580个氨基酸。利用MAGE 5.0软件绘制PPV系统发育进化树,结果显示,PPV毒株可以划分为4组,我国的PPV毒株主要集中在A和B两组,仅JY毒株位于C组;PPV VP2蛋白3维结构显示,氨基酸变异的多态性位点主要集中在病毒衣壳蛋白的表面。以上遗传进化关系表明,PPV HN-2011株是我国PPV的流行毒株。

According to the VP2 gene sequence of porcine parvovirus（PPV） published in GenBank database,a pair of specific primers was designed and synthesized.The full length VP2 gene of ppv HN-2011 isolate was amplified by PCR and then cloned into pMDl8-T vector.As expectedly,the complete nucleotide sequence of VP2 gene was 1 740 nt and coded 580 amino acids.The phylogenetic tree showed that PPV isolates were divided into four groups,and the Chinese isolates were clustered in groups A and B,except for the JY strain in group C.Mapping polymorphic sites of protein data onto the three-dimensional（3D） structure of PPV VP2 revealed that almost all substitutions were located on the external surface of the viral capsid.According to the phylogenetic relationship of the VP2 gene,the isolate HN-2011 was a prevalent PPV strain in China.