狂犬病病毒磷蛋白在杆状病毒中的表达及鉴定  

Expression and Characterization of Rabies Virus Phosphoprotein in Baculovirus

作　　者：郑光来 卢晓冉 张永武[1] 张静远[1] 陈腾[1] 王东方[1] 严妍[1] 张守锋[1] 扈荣良[1] 

机构地区：[1]军事医学科学院军事兽医研究所吉林省人兽共患病预防与控制重点实验室,吉林长春130122

出　　处：《动物医学进展》2016年第5期26-29,共4页Progress In Veterinary Medicine

基　　金：国家863计划项目(2011AA10A212)

摘　　要：采用RT-PCR方法扩增出狂犬病病毒BD06株的磷蛋白(P)基因片段,将其插入杆状病毒穿梭载体pFastBac 1获得的pFastBac 1-P重组质粒,转化E.coli DH10Bac成功构建出杆状病毒表达质粒Bacmid-P,用脂质体介导Bacmid-P转染Sf9细胞获得重组杆状病毒(AcMNPV-P)。经SDS-PAGE、Western blot和直接免疫荧光分析鉴定,BD06-P蛋白在杆状病毒中成功表达,且具有良好的反应原性,为进一步研究狂犬病病毒磷蛋白的结构和功能奠定了基础。The phosphoprotein gene of rabies virus BD06 stain was amplified by RT-PCR and cloned into the baculovirus shuttle vector pFastBac 1,then the recombinant vector pFastBac 1-P was transformed into E. coli DH10Bac to obtain the recombinant expression plasmid Bacmid-P. The recombinant baculovirus AcM- NPV-P was obtained by transfecting the Bacmid-p into Sf9 cells. The expressed recombinant phosphopro- rein was identified by SDS-PAGE,Western blot and direct immunofluoresence assay. The results showed that the BD06-P was expressed in recombinant baculovirus successfully and had a good antigenicity, it would be potentially used for further study on the structures and functions of the phosphoprotein of rabies virus.

关 键 词：狂犬病病毒 杆状病毒表达系统 磷蛋白 

分 类 号：S852.655[农业科学—基础兽医学]