为了获得瑞斯胆量的 P8 蛋白质,与生物活动使病毒(RGDV ) 相形见绌,它的外部上衣蛋白质基因 S8 在 Spodoptera frugiperda (Sf9 ) 被表示用 baculovirus 表示系统的昆虫房间。S8 基因是进 pFastBac 的 subcloned ? 1 向量,生产 recombinant baculovirus 转移向量 pFB-S8。在转变以后, pFB-S8 被介绍进能干的房间(E。coli DH10Bac ) 包含梭向量, Bacmid,产生 recombinant bacmid rbpFB-S8。在被 recombinant baculovirus rvpFB-S8 在感染的不同复合感染以后, Sf9 房间在不同时间被收集并且由 SDS 页,西方的弄污和 immunofluorescence 显微镜学分析了。P8 蛋白质的表示水平在在 Sf9 房间的 transfection 以后的 4872 h 之间是最高的。Immunofluorescence 显微镜学证明 RGDV 的那 P8 蛋白质在 Sf9 房间的细胞质形成了有小斑点的结构。
To obtain the P8 protein of Rice gall dwarf virus (RGDV) with biological activity,its outer coat protein gene S8 was expressed in Spodoptera frugiperda (Sf9) insect cells using the baculovirus expression system.The S8 gene was subcloned into the pFastBacTM1 vector,to produce the recombinant baculovirus transfer vector pFB-S8.After transformation,pFB-S8 was introduced into the competent cells (E.coli DH10Bac) containing a shuttle vector,Bacmid,generating the recombinant bacmid rbpFB-S8.After being infected by recombinant baculovirus rvpFB-S8 at different multiplicities of infection,Sf9 cells were collected at different times and analyzed by SDS-PAGE,Western blotting and immunofluorescence microscopy.The expression level of the P8 protein was highest between 48-72 h after transfection of Sf9 cells.Immunofluorescence microscopy showed that P8 protein of RGDV formed punctate structures in the cytoplasm of Sf9 cells.