中文摘要：

[目的]为获得家蚕二分浓核病毒（Bm BDV）VD1-ORF4编码的可溶性蛋白,对该序列在两类表达系统中进行表达。[方法]在原核系统中,将靶基因克隆入载体p MAL-c2X中,使麦芽糖标签序列与靶基因5’端融合,将融合后的重组质粒转化大肠杆菌BL21,进而对其进行IPTG诱导,对诱导后的表达产物进行Western blotting分析;在真核表达系统中,以笔者实验室改造的Ac-Bacmid为分子载体,通过转座,将多角体启动子控制的VD1-ORF4表达盒定点插入到载体中,在脂质体的介导下,将整合型重组杆粒转染Sf-9细胞,将转染上清感染Sf-9细胞,并对其总蛋白进行Western blotting分析。[结果]在两类表达系统中,都只鉴定到Bm BDV VD1-ORF4部分序列的表达,其融合表达产物大小都约70 kDa。[结论]获得Bm BDV VD1-ORF4截短蛋白,为其功能研究奠定基础。

英文摘要：

[Objective]To achieve the soluble protein encoded by Bombyx mori bidensovirus（Bm BDV） VD1-ORF4,two different expression system were used to express Bm BDV VD1-ORF4 in this study.[Methods]In eukaryotic expression system,Bm BDV VD1-ORF4 was cloned into p MAL-c2 X vector for the expression of target protein fusion with maltose binding protein（MBP） tag.Meanwhile,Bm BDV VD1-ORF4 under the control of the polhedrin was transposed into a modified Ac-Bacmid,and the resulting recombinant Ac-Bacmid was transfected into Sf-9 cells to generate recombinant virus.The virus supernatant was harvested and used to infect Sf-9 cells for the expression of target protein.The total protein extracts from the E.coli and Sf-9 cells infected with recombinant virus were subjected to Western blotting analysis,respectively.[Results]The result showed that a truncated protein with molecular weight of 70 kDa was examined.[Conclusion]Only partial sequence of Bm BDV VD1-ORF4 was expressed in E.coli or Sf-9 cells.