中文摘要：

为优化家蚕杆状病毒表达系统,提高外源基因的表达产量。文中通过同源重组技术,用串联的氯霉素基因（Cm）表达盒和绿色荧光蛋白基因（egfp）表达盒将其替换,从而获得Chitinase和Cystein Protease两个基因缺失的家蚕杆状病毒载体。通过转座,将多角体启动子控制的家蚕二分浓核病毒（Bm BDV）ns1基因表达盒,定点插入到改造后的该分子载体中。将重组载体转染Bm N细胞,获得能表达家蚕二分浓核病毒（Bm BDV）NS1的缺失型重组病毒;另外,将多角体启动子控制的ns1基因转座到野生型Bm-bacmid中,获得能表达Bm BDV NS1的野生型重组病毒。将这两种病毒分别皮下注射家蚕,对感染后的家蚕血液中NS1表达水平进行比较,发现缺失Chitinase和Cystein Protease重组病毒感染的家蚕血液中,NS1的表达量是对照组的3倍,从而建立了一种高效表达可溶性NS1蛋白的方法,为靶蛋白的结构与功能研究奠定基础。

英文摘要：

To improve the expression of heterologous genes using baculovirus expression system, we constructed a novel shuttle vector based on the Bm-Bacmid. In the Bm-Bacmid, partial sequences of Chitinase and Cystein Protease were replaced with a tandem cassette of Cm and egfp through homologous recombination. Bombyx mori bidensovirus（Bm BDV） ns1 under the control of polyhedrin promoter was inserted into the modified Bm-bacmid by transposition. For comparison, Bm BDV ns1 under the control of polyhedrin promoter was also cloned in the wild type Bm-bacmid. The resulting Bm-bacmids were transfected into the cultured Bm N cells to prepare recombinant virus to infect silkworms for expression of Bm BDV ns1. Total proteins of hemocyte from infected silkworms were subjected to Western blotting and ELISA analysis. The yield of Bm BDV NS1 with the modified vector was three times as much as that with the unmodified vector. The method to improve the yield of Bm BDV NS1 in silkworms will facilitate the function and three-dimensional structure study of Bm BDV NS1.