兔出血症病毒衣壳蛋白P区二聚体的表达及其与受体结合能力分析  

Expression of the P Dimer of Rabbit Hemorrhagic Disease Virus Capsid Protein and Analysis of Its Binding Ability to Receptor

作　　者：胡波[1] 范志宇[1] 王芳[1] 魏后军[1] 宋艳华[1] 仇汝龙 徐为中[1] 薛家宾[1] 

机构地区：[1]江苏省农业科学院兽医研究所.农业部兽用生物制品工程技术重点实验室.国家兽用生物制品工程技术研究中心,南京210014

出　　处：《畜牧兽医学报》2016年第2期325-330,共6页ACTA VETERINARIA ET ZOOTECHNICA SINICA

基　　金：现代农业产业技术体系建设专项资金(CARS-44);江苏省农业科技自主创新资金项目[CX(13)5029]

摘　　要：为检测兔出血症病毒(RHDV)衣壳蛋白(VP60)P区形成二聚体的能力及其与HBGAs受体结合的能力,作者以pMD19T-VP60为模板扩增包含铰链区H的P区基因(HP)并克隆至原核表达载体pET28a(+)中,转化大肠杆菌BL21(DE3)菌株,经IPTG诱导表达,获得HP蛋白。经SDS-PAGE和Western blot检测,结果显示,HP蛋白主要以包涵体的形式高效表达,经HisTrap亲和柱纯化后可形成二聚体结构。通过HBGAs受体结合试验表明,P区与完整病毒衣壳相似,能与唾液中的HBGAs受体发生结合。本研究为进一步开展RHDV衣壳蛋白与受体的相互作用研究奠定基础。Rabbit hemorrhagic disease virus（RHDV）is a member of the Caliciviridae family（Lagovirus genus）.Similar to human norovirus（Caliciviridae,Norovirus genus）,RHDV binds histo-blood group antigens（HBGAs）and this is thought to be important for infection.The aim of this study was to determine whether the P domain of RHDV capsid protein（VP60）could form dimers and to analyze its binding ability to HBGAs receptor.The P domain gene containing hinge（HP）was amplified and cloned into pET-28a（＋）expression vector and introduced into Escherichia coli BL21（DE3）.The recombinant HP protein,confirmed by SDS-PAGE and Western blot,was effectively expressed in form of inclusion bodies by inducing with IPTG.The recombinant protein was then purified with HisTrap affinity chromatography,which could form dimers after purification.The HBGAs binding assay showed that the P domain bound H type HBGA with the same patterns as those of the intact viral capsids.Further structural studies with P domain are needed in order to better understand the HBGA binding mechanisms and virus-receptorinteraction.

关 键 词：兔出血症病毒 衣壳蛋白 P区 组织血型抗原 受体结合 

分 类 号：S858.291[农业科学—临床兽医学] S852.659.6[农业科学—兽医学]