番鸭细小病毒浙江分离株VP基因的克隆与序列分析

Cloning and Sequence Analysis of VPgene of Muscovy Duck Parvovirus Zhejiang Isolate

为明确番鸭细小病毒结构蛋白VP的特征,本研究运用PCR从已分离鉴定的番鸭细小病毒浙江分离株(MDPV-ZJ)中分段扩增出VP基因,并对其进行克隆测序和分析。结果表明,番鸭细小病毒浙江分离株VP基因全长为2 199bp,编码732个氨基酸。所编码的包膜蛋白大小为81.32ku、理论等电点为6.59、不稳定系数为37.49、亲水性平均系数为-0.667,属于亲水性稳定类蛋白,该编码的结构蛋白没有信号肽,为非分泌蛋白。根据VP基因特征从遗传进化上可将MDPV分为2个大的基因型:经典型和基因重组型,经典型MDPV可以进一步划分为台湾亚群、大陆亚群和欧洲群,MDPV各分离株在遗传进化上存在明显的地域性。本试验中MDPV浙江分离株株处于MDPV大陆亚群。

In order to enrich the biological characteristics of VP gene from Muscovy duck parvovirus Zhejiang isolate（MDPV-ZJ）,the target VP gene fragments were amplified by PCR method with specific primers,then the obtained PCR products were cloned and sequenced.The bioinformatics analysis of VP gene of MDPV-ZJ was conducted.The results revealed that MDPV-ZJ VP gene was 2 199 bp in length,coding an open reading frame（ORF）with 732 amino acids.The molecular weight,theoretical isoelectric point,instability index and grand average of hydropathicity of MDPV-ZJ VP gene were 81.32 ku,6.59,37.49and-0.667,respectively,and with no signal peptide.The genetic evdution ary tree based on the VP gene showed that the MDPV isoaltes contains two groups：The typical MDPV and recombinant MDPV;Also,the typical MDPV could be divided into three branches：Taiwan Residents branch,Maniland China branch and Euro branch,which existed obvious regional genetic evolution relationship.In this assay MDPV-ZJ was belonged to MDPV Maniland China branch.