抗猪伪狂犬病病毒gH抗体的制备及gH在感染细胞中表达分析  

Preparation of antibodies against pseudorabies virus (PRV) gH and detection of gH in PRV infected cell

作　　者：刘庆伟[1,2] 邱亚峰[1] 王晓杜[1] 郭林[1] 刘超[1] 沈阳[1] 李向东[1] 单虎[2] 马志永[1] 

机构地区：[1]中国农业科学院上海兽医研究所兽医公共卫生实验室,上海200241 [2]青岛农业大学动物科技学院,山东青岛266109

出　　处：《中国预防兽医学报》2011年第5期383-387,共5页Chinese Journal of Preventive Veterinary Medicine

基　　金：国家自然科学基金青年基金(30700593);中央级公益性科研院所基本科研业务费专项资金项目

摘　　要：为检测伪狂犬病病毒(PRV)在体外细胞中糖蛋白H(gH)的表达情况,本研究构建原核表达重组质粒pET-gHN660,并在大肠杆菌中诱导表达重组蛋白,纯化的gH重组蛋白免疫实验动物制备抗PRVgH抗体,经westernblot和IFA检测到病毒感染细胞中gH蛋白的表达,病毒感染细胞后表达的gH蛋白大小为95ku,定位于细胞浆中,gH蛋白在病毒感染细胞4h可以检出,随PRV的复制gH蛋白表达增加,gH蛋白可以作为PRV复制的指示蛋白。本研究利用制备的抗体分析感染细胞中gH蛋白的表达情况,初步探讨感染细胞中PRV的复制,为PRV和宿主相互作用的研究奠定基础。To analysis the expression of glycoprotein H （gH） in pseudorabies virus （PRV） infected cell, partial domain of PRV gH was expressed in E. coli and purified to prepare the anti-PRV gH antibodies in rabbit. gH protein expressed in PRV infected cell were detected by western blot and immunofluorescence. The results showed that the molecular weight of PRV gH was 95 ku and expressed at 4 hours post infection, mainly located in the cytoplasm, and accumulated as PRV replication, PRV gH would be a potential indicator protein of PRV replication in infected cells. The expression of PRV gH were analysised using anti-PRV gH antibodies, and the PRV replication was discussed.

关 键 词：伪狂犬病病毒 GH基因 抗PRVgH抗体 

分 类 号：S852.65[农业科学—基础兽医学]