EHV-1皮层蛋白VP22生物信息学分析及亚细胞定位研究  

Bioinformatics analysis and subcellular localization of tegument protein VP22 of Equine herpesvirus type 1

作　　者：王世民[1] 王浩 阿依古丽·艾麦尔尼亚孜 郝翠兰[1] 姚刚[1] WANG Shimin;WANG Hao;Aygul Amarniyazi;HAO Cuilan;YAO Gang(College of Veterinary Medicine,Xinjiang Agricultural university,Urumqi 830052,China)

机构地区：[1]新疆农业大学动物医学学院

出　　处：《黑龙江畜牧兽医》2018年第21期32-36,258,259,共7页Heilongjiang Animal Science And veterinary Medicine

基　　金：新疆维吾尔自治区科技基础条件平台建设项目(PT1613)

摘　　要：为了研究马疱疹病毒（EHV-1）皮层蛋白VP22的生物学功能,试验通过克隆测序获得VP22基因并构建系统进化树,采用生物信息学软件分析其生物学特征,并将其编码基因的PCR扩增产物亚克隆至p DsRed2-N1真核表达载体,构建重组质粒pDsRed-VP22,研究VP22在BHK-21细胞中定位。结果表明：EHV-1新疆本地株的VP22由304个氨基酸组成,其序列与欧洲分离株Ab4的同源性为100%,与其他分离株的同源性为97%-99%; VP22是一种无信号肽、跨膜区及核定位信号（NLS）的磷酸化蛋白质,具有1个核输出信号（NES）序列及14个蛋白结合位点; VP22的二级结构主要由α-螺旋及β-折叠构成,三级结构及表面电荷分布情况显示该蛋白质正反两面的电荷分布差异较大;生物学功能预测显示EHV-1 VPP22是一种多功能蛋白质;此外,试验成功构建出重组质粒p DsRed-VP22,并证实EHV-1 VP22定位于BHK-21细胞的内质网-高尔基体（ER-Golgi）膜系附近。The aim of the present study was to investigate the biological function of tegument protein VP22 encoded by Equine herpesvirus type 1（EHV-1）. The VP22 gene was cloned and sequenced and the phylogenetic tree was constructed. Some bioinformatics software were used to analyze its biological characteristics. Then the PCR products of VP22 were cloned into the eukaryotic expression vector p DsRed2-N1 to construct the recombinant plasmid pDsRed-VP22,so as to study localization of VP22 in BHK-21 cells. The results showed that the VP22 protein of EHV-1 strain isolated locally was composed of 304 amino acids,and its sequence was 100% homologous to that of the Ab4 isolate,97%-99%other isolates. VP22 is a kind of phosphorylated protein that possesses a nuclear export signal（NES） and 14 protein binding sites,but no signal peptide,trans-membrane domain and nuclear locaization signal（NLS）. The secondary structure of VP22 is mainly constituted by α-Helix andβ-sheet. The 3 D structure and the charge calculation of VP22 showed that the distribution of surface charge differed from the two opposite surfaces. The pDsRed-VP22 plasmid was constructed successfully and proved that the VP22 was located near the endoplasmic reticulum-Golgi（ER-Golgi） network of BHK-21 cells.

关 键 词：马疱疹病毒 皮层蛋白 二级囊膜 亚细胞定位 系统发育树 分子模拟 

分 类 号：S852.659.1[农业科学—基础兽医学] Q51[农业科学—兽医学]