GP64表面展示元件与组Ⅱ HaSNPV出芽病毒的融合  

GP64 surface display elements assembled to the HaSNPV budded virus of group Ⅱ baculovirus

作　　者：刘永[1] 于峰[1] 曹青[1] 徐琳琳[1] 陈修文[1] 王文兵[1] 

机构地区：[1]江苏大学生命科学研究院,镇江212013

出　　处：《生物学杂志》2012年第6期14-18,共5页Journal of Biology

基　　金：国家重点基础研究发展计划"973"项目(编号:2005CB121005)

摘　　要：利用杆状病毒组ⅠAcMNPV(Autographa californica multiple nucleopolyhedrovirus)GP64信号肽(GP64SP),C-末端跨膜区(GP64CTD)及报告基因绿色荧光蛋白(eGFP)构成的表面展示系统(gp64sp-egfp-gp64ctd),同源重组到组ⅡHaSNPV的多角体基因位置,筛选重组病毒。通过激光共聚焦观察,HaSNPV重组病毒感染Hz-AM1细胞后绿色荧光分布在质膜,表明GP64SP可以引导表达产物趋向于细胞膜。病毒感染后,收集并纯化重组的出芽病毒(BV),经Western blot检测,eGFP存在于重组病毒BV中。结果表明,组ⅠGP64表面展示系统中的重要元件GP64SP和GP64CTD能够装配到组ⅡHaSNPV BV上,可以用于组ⅡNPV表面展示目的蛋白。The GP64 signal peptide（GP64SP） and the C-terminal transmembrane domain（GP64CTD） of AcMNPV were used as surface display system in group Ⅰ baculovirus.In this study,the surface display structure（gp64sp-egfp-gp64ctd） was inserted into the polyhedron position of HaSNPV（Helicoverpa armigera single nucleopolyhedrovirus）,and the recombinant virus was selected.Green fluorescence was observed in the cellular membrane of the infected Hz-AM1 cells by confocal microscopy.Western blot analysis using an eGFP-antibody also showed that the eGFP-fused protein was present in the budded viruses.These data implied that GP64 surface display elements GP64SP and GP64CTD of group Ⅰ baculovirus also acted in group Ⅱ baculovirus.

关 键 词：GP64 组Ⅱ 表面展示 HASNPV 出芽病毒 

分 类 号：Q78[生物学—分子生物学]