蓝舌病病毒衣壳蛋白VP2、VP5、VP7酵母双杂交诱饵质粒的构建及鉴定  

Construction and Identification of Yeast Two-hybrid Bait Plasmids of Capsid Proteins VP2,VP5 and VP7 of Bluetongue Virus

作　　者：张继凯[1] 孙恩成[1] 杨涛[1] 徐青元[1] 吕爽[1] 王海秀[1] 张沁[1] 吴东来[1] 

机构地区：[1]中国农业科学院哈尔滨兽医研究所,兽医生物技术国家重点实验室,农业部兽医公共卫生重点开放实验室,哈尔滨150001

出　　处：《中国畜牧兽医》2016年第1期39-44,共6页China Animal Husbandry & Veterinary Medicine

基　　金：国家自然科学基金(31302065);中国博士后科学基金(2013M540172);国家科技支撑计划(2013BAD12B05);公益性行业(农业)科研专项(201203056);新型重大动物疫苗与诊断试剂创制及生产工艺创新(2011AA10A213);中央级公益性科研院所基本科研业务费(0302015008)

摘　　要：蓝舌病病毒(bluetongue virus,BTV)的结构主要由3层衣壳蛋白组成,其中VP2、VP5蛋白构成了BTV的外层衣壳,VP7蛋白构成了BTV的中间衣壳,最内层衣壳则由VP3蛋白构成。VP2、VP5及VP7蛋白在BTV侵染宿主细胞的过程中起着非常重要的作用。为了研究BTV与宿主细胞相互作用的分子机制,本研究将BTV的VP2、VP5、VP7基因分别克隆到pGBKT7载体中,成功构建了pGBKT7-VP2、pGBKT7-VP5与pGBKT7-VP7 3个诱饵质粒,且通过自激活和毒性验证,证明所构建的3个质粒均无自激活作用,对酵母细胞无毒性作用。本研究为今后利用酵母双杂交筛选VP2、VP5、VP7蛋白中与宿主细胞相互作用的蛋白做好了铺垫,为深入研究BTV与宿主细胞的相互作用奠定了基础。The structure of bluetongue virus（BTV）was consisted of three layers of capsid proteins,VP2 and VP5proteins consisted the outer capsid of BTV,VP7 protein consisted the middle capsid of BTV,VP3 consisted the inner capsid of BTV.When BTV infected host cells,VP2,VP5 and VP7proteins of BTV played important roles in the process of infecting host cells.In order to study the molecular mechanism of interaction between BTV and host cells,we cloned VP2,VP5 and VP7genes into pGBKT7 vector,three recombinant bait plasmids pGBKT7-VP2,pGBKT7-VP5 and pGBKT7-VP7 were successfully constructed,and then the self-activation and toxicity of the bait plasmids were tested.The results showed that three bait plasmids all had no self-activation and toxicity to yeast cells.This research made a steppingstone for the screening of host-cell protein interacting with VP2,VP5 and VP7proteins using yeast two-hybrid system,and laid a foundation for investigating the interaction between BTV and its host cells.

关 键 词：蓝舌病病毒 诱饵质粒 自激活作用 毒性作用 

分 类 号：Q78[生物学—分子生物学]