ClanGV的PIF0与其他经口感染因子间的互作研究  

Identification of Interactions between PIF0 and the other PIFs of ClanGV

作　　者：梁振普[1] 吴慧[1] 张小霞[1] 张亲[1] 王彩平[1] 刘雅静[1] 张俊庆[1] 桑思瑶 

机构地区：[1]河南农业大学生命科学学院,郑州450002

出　　处：《病毒学报》2017年第2期245-252,共8页Chinese Journal of Virology

基　　金：国家自然科学基金(项目号:31570151);题目:苜蓿银纹夜蛾核多角体病毒AC146蛋白在病毒粒子发生中的功能研究;国家自然科学基金(项目号:31272094);题目:棉铃虫围食膜上ENHANCIN酶解靶标的鉴定及其功能位点解析

摘　　要：本文旨在通过酵母双杂交技术研究杨扇舟蛾颗粒体病毒(ClanGV)的经口感染因子PIF0和其他经口感染因子PIF1~PIF6之间的互作关系。首先根据ClanGV的基因组序列信息,利用在线软件TMHMM Server v.2.0对PIF0~PIF6进行跨膜区分析,确定需要扩增的片段。然后以诱饵载体pGBKT7和捕获载体pGADT为出发载体,构建了PIF0的重组诱饵载体(B0)和PIF1~PIF6的重组捕获载体(A1~A6)。自激活实验证明,重组诱饵载体B0没有自激活特性,可以进一步开展蛋白的互作研究。酵母双杂交结果证明,PIF0作为诱饵载体时,可以与PIF3和PIF5的重组捕获载体发生蛋白相互作用,与PIF1、PIF2、PIF4和PIF6都没有互作现象。本研究结果将有助于揭示PIF0在ClanGV经口感染过程中的功能,并可为阐明ClanGV的经口感染机制以及开发新型病毒杀虫剂和增效剂奠定基础。During the infection cycle,baculoviruses produce two virion phenotypes：occlusion derived virus（ODV）and budded virus（BV）.Infection by the ODV of the baculovirus is mediated by highly conserved per os infectivity factors（PIFs）.The Clostera anachoreta granulovirus is a member of Betabaculovirus,and has been used for C.anachoretacontrol in China.We wished to identify the interaction between PIF0 and the other PIFs of ClanGV（a member of Betabaculovirus）by yeast two-hybrid（Y2H）screening.Potential transmembrane domains were screened by TMHMM software according to the genome sequence of PIF0-PIF6 of ClanGV to determine the target sequence.The pGADT7（prey）recombinant vector of PIF0 and pGBKT7（bait）recombinant vectors of PIF1-PIF6 were constructed by homologous recombination.The recombinants were named B0 and A0-A6,respectively.Self-activation experiments suggested that B0 did not exhibit self-activation.Y2 Hscreening suggested that PIF0 is a bait vector that interacts only with PIF3 and PIF5,and not with PIF1,PIF2,PIF4 or PIF6.Our study will help to reveal the function of PIF0 during oral infection by ClanGV.It may also shed light on the mechanism of oral infection,and aid development of novel virus pesticides and synergistic agent.

关 键 词：杨扇舟蛾颗粒体病毒 经口感染因子 酵母双杂交 蛋白互作 

分 类 号：S763.12[农业科学—森林保护学]