苜蓿银纹夜蛾多核衣壳核型多角体病毒AcMNPVP78/83蛋白初步研究  

FunctionalAnalysis ofAcMNPV P78/83 Protein

作　　者：聂英超[1] 方明刚[1] 王倩[1] 余泽华[2] 陈新文[1] 

机构地区：[1]中国科学院武汉病毒研究所分子病毒学实验室,湖北武汉430071 [2]华中师范大学生命科学学院昆虫所,湖北武汉430079

出　　处：《中国病毒学》2004年第5期498-503,共6页Virologica Sinica

基　　金：国家973项目(2002CCCA028002003CB114202);国家自然科学基金杰出青年基金(30325002)

摘　　要：AcMNPVORF9编码病毒核衣壳蛋白P78/83,该蛋白在宿主细胞内以磷酸化和去磷酸化两种形式存在,能够与细胞骨架成分肌动蛋白相互作用,序列分析表明其具有与WASP蛋白类似的结构,推测可能在病毒粒子的包装、运输等过程中起重要作用。本文利用Bac-to-Bac系统构建了P78/83与绿色荧光蛋白融合表达的重组AcMNPV,激光共聚焦显微镜观察表明,重组病毒感染Sf21细胞12h后绿色荧光主要集中分布于细胞质中,24h及以后绿色荧光主要集中分布于细胞核中。感染试验表明,超表达P78/83对病毒的生长无明显的影响。Autographa californica nucleopolyhedrovirus (AcMNPV) is the typical member of nucleopolyhedrovirus of Baculoviradae, a circled and double stranded large DNA virus. It’s also one of the most widely studied baculoviruses that turns out to be environment sound pesticide and a powerful vector of foreign protein expression as well as a promising vector of gene therapy. Revealing the molecular mechanism of the interaction plays a key role in the better understanding of viral replication, package, transportation and application of baculovirus as a gene therapy vector. Sequence analysis found that the P78/83 product of AcMNPV ORF9 shares the common motifs with WASP proteins indicating that P78/83 might involve in baculovirus transportation. Trying to understand P78/83 function, we constructed a recombinant virus vAc-orf9-gfp with the Bac-to-Bac system, within which AcMNPV ORF9 was fused with enhanced green fluorescence protein (EGFP). Sf21 cells were infected with the recombinant virus and Laser Confocal Microscopy was used to determine the subcellular location of P78/83. Green fluorescences showed major distribution in cell plasm a after 12h infection, and transfered into nucleus at 24 hpi. This means that P78/83 is located in the nucleus. Infection assay in vitro showed that overexpression of P78/83 had no effect on virus growth and viral assembly.

关 键 词：苜蓿银纹夜蛾多核衣壳核多角体病毒 绿色荧光蛋白 P78/83蛋白 

分 类 号：Q939.4[生物学—微生物学]