中文摘要：

构建亚非马蜂（Polistes hcbracus）溶血肽基因重组转移载体pBacHT-GFPTPhMT，转化含穿梭载体Bacmid的受体菌Eschcrichia coil（DH10Bac）中，得重组穿梭载体Bacmid-GFPTPhM,再用Lipofectin介导其DNA转染粉纹夜蛾（Trichoplusia ni）细胞系Tn。SDS-PAGE分析表明，感染Bacmid-GFPTPhM的Tn-581-4细胞的表达产物在约为34kD处出现特异性条带，表达量约占细胞总蛋白的3％。Western blot显示表达产物具有良好的免疫活性。感染Bacmid-GFPTPhM的细胞表达时相动态分析进一步表明，与增强型绿色荧光基因融合的亚非马蜂溶血肽基因的重组病毒已在昆虫细胞中进行了成功地表达，感染后72h表达量可达最高水平。

英文摘要：

A cDNA fragment encoding melittin from the venom gland of female Polistes hebraeus was cloned into pBacHTeGFPT to construct the recombinant donor plasmid pBacHT-GFPTPhM. The donor plasmid was then transformed into Escheriehia coil （DH10Bac） to form the recombinant, Bacmid-GFPTPhM. Baemid-GFPTPhM DNA was further transfected into Tn-5B1-4 cells of the cabbage looper, Trichoplusia ni, mediated by Lipofectin. SDS-PAGE result showed the expressed protein was about 34 kD. The thin layer scanning showed that the expressed fusion protein GFPT-PhM was accumulated up to about 3 % of the total cell protein. The recombinant product was confirmed by Western blot using the His-Tag monoelonal antibody and a rabbit derived polyelonal antibody against melittin, respectively. The eytopathie effect of Tn-5B1-4 cells after infection of Baemid-GFPTPhM at different times was inves- tigated and the result indicated that the recombinant protein was expressed to the highest level in Tn-5B1-4 cells 72 h post infection.