家蚕细胞中过表达家蚕核型多角体病毒核衣壳蛋白VP39抑制BmNPV的增殖  

Overexpression of Bombyx mori nucleopolyhedrovirus nucleocapsid protein VP39 inhibits the proliferation of Bm NPV in Bm N-SWU1

作　　者：胡楠[1] 董战旗 陈婷婷[1] 潘敏慧[1,2] 

机构地区：[1]西南大学家蚕基因组生物学国家重点实验室,重庆400716 [2]西南大学农业部家蚕功能基因组和生物学重点实验室,重庆400716

出　　处：《昆虫学报》2015年第11期1222-1228,共7页Acta Entomologica Sinica

基　　金：国家自然科学基金项目(31272505;31172269;31472152);中央高校基本业务费专项资金(XDJK2015D002)

摘　　要：【目的】家蚕Bombyx mori核型多角体病毒(Bm NPV)核衣壳蛋白VP39为病毒装配所必需。本研究旨在初探VP39在病毒侵染家蚕细胞过程中的功能及特征,以期为家蚕抗病毒研究提供研究基础。【方法】本研究通过构建原核表达载体,诱导原核表达得到多克隆抗体,以Western blot验证VP39表达时相;构建真核表达载体,转染细胞后以免疫荧光手段观测VP39表达定位及影响病毒增殖现象。【结果】制备了VP39多克隆抗体。VP39在病毒感染后大量定位于家蚕细胞核,部分定位于胞质,而过表达的VP39定位于家蚕细胞胞质;过表达VP39后抑制Bm NPV感染家蚕细胞。【结论】在Bm N-SWU1细胞中过表达VP39会影响Bm NPV的扩散,导致Bm NPV感染细胞数目大量减少。该结果为VP39调控宿主与病毒的相互作用提供了新的思路。[Aim]Bombyx mori nucleopolyhedrovirus nucleocapsid protein VP39 is necessary for virus assembly. This study aims to explore the functions and characteristics of VP39 in the infection process of virus in silkworm cells and so to make a contribution to the antivirus research of the silkworm. [Methods]The prokaryotic expression vector was constructed to induce and produce the VP39 polyclonal antibody and to test the time course of protein expression by Western blot. The location of VP39 and the effect of VP39 on Bm NPV proliferation in silkworm cells were observed by transfection of eukaryotic expression vector p IZ-vp39-V5 / His and immunofluorescence. [Results ] We obtained the VP39 polyclonal antibody,and verified that VP39 was mostly localized in cells nucleus areas during Bm NPV infection,but just localized in the cytoplasm when overexpressed. The overexpressed VP39 inhibited the proliferation of Bm NPV in Bm N-SWU1. [Conclusion ]The results suggest that the overexpression of VP39 in Bm NSWU1 cells may inhibit the proliferation of Bm NPV,thus offering a new way for exploring the regulation of VP39 in the interaction between host cell and virus.

关 键 词：家蚕 核型多角体病毒 VP39 抗病毒 病毒增殖 亚细胞定位 

分 类 号：Q965.8[生物学—昆虫学]