中文摘要：

目的：对家蚕核型多角体病毒（BmNPV）囊膜蛋白P74膜外肽段与枯草芽胞杆菌（Bacillus subtilis）CotC与进行融合表达,制备表面展示有P74蛋白的重组芽孢,为深入研究该重组芽孢的功能提供基础。方法：将CotC基因与BmNPV P74膜外编码序列进行融合,构建表达CotC-P74融合蛋白的重组质粒pJS700-p74。通过双交换使该重组质粒中的CotC-P74表达盒整合到枯草芽胞杆菌染色体淀粉酶基因位点,并对发生同源重组后的菌株进行筛选和PCR鉴定。结果：PCR结果表明CotC-P74成功地整合到枯草芽孢杆菌基因组上,通过作者实验室制备的P74多抗对诱导后的重组芽孢衣壳总蛋白进行Western blot分析,结果在49 kDa位置处能杂交到一条特异的蛋白带。结论：P74蛋白胞外肽段成功地展示在枯草芽胞杆菌芽胞表面。

英文摘要：

Objective： To display P74 on the surface of Bacillus subtilis （ B. subtilis ） spores for further functional analysis. Method： A recom- binant integrative plasmid named as pJS700 -p74,carrying cotC- p74 under the control of the cotC promoter, was constructed. Competent cells B. subtilis 168 （trp - ）were transformed with recombinant pJS700 -p74, which generated the resulting amylase inactivated mutant for a double crossover. Result：The disruption of amyE and the insertion of cotC -p74 in the amyE locus were confirmed by PCR, and a specif- ic hand about 49 kDa was detected in the recombinant spores extracts using P74 antibodies. Conclusion：P74 was displayed on B. subtilis spore surface successfully.