【摘要】 P10蛋白是杆状病毒感染细胞后在极晚期高度表达的两个蛋白之一。本文通过构建p10缺失的重组棉铃虫病毒来研究P10的功能。利用λ噬菌体Red重组系统介导的同源重组,在大肠杆菌BW25113中,用含有40bp同源臂的氯霉素抗性基因替换了棉铃虫病毒(HaSNPV)细菌人工染色体HaBacHZ8上的p10基因,然后利用Bac-to-Bac系统把多角体基因和绿色荧光蛋白基因转移到含有缺失p10的HaBacHZ8上,构建了p10缺失的重组HaBacΔp10-PH-eGFP。重组病毒的生长曲线和生物测定结果表明,缺失p10基因对病毒复制和毒力无显著影响,但电镜观察结果显示,P10的缺失会影响多角体囊膜的包装。更多还原

【Abstract】 p10 is one of the two very late genes that are highly expressed in baculovirus infected cells. We have studied the function of P10 by constructing a p10 deletion recombinant of the Helicoverpa armigera singly-enveloped nucleopolyhedrovirus (HaSNPV). The p10 gene of HaSNPV bacterial artificial chromosome (HaBacHZ8) was replaced with a DNA fragment containing a chloramphenicol antibiotic gene and a 40bp flanking region using the λ phage Red recombination system in E. coli BW25113. Polyhedrin and gfp genes were then translocated to the p10 deleted HaBacHZ8 using a Bac-to-Bac system generating the recombinant bacmid HaBacΔp10-PH-eGFP. Virus growth curves and bioassays indicated that deletion of p10 did not impair the infectivity of virus in vitro and in vivo. However, electronic microscopic analysis revealed that deletion of P10 resulted in the fragmentation of polyhedral envelope.更多还原