Wu Wu-Wei, Wang Jing-Wen, Xie Fei, Long Qing-Xin, Wang Xun-Zhang
State Key Laboratory for Biocontrol, Zhongshan University, Guangzhou 510275, China.
Sheng Wu Hua Xue Yu Sheng Wu Wu Li Xue Bao (Shanghai). 2003 Sep;35(9):834-40.
The p74 gene of Autographa californica multicasid nucleopolyhedrovirus (AcMNPV) bacmid was knockouted and substituted by the p74 gene of Spodoptera litura multicapsid nucleopolyhedrovirus (SpltMNPV), using RecA-mediated homologous recombination in the E. coli. No selection marker, which might influence the expression and function of p74 gene, was left in the modified p74 locus. The promoter of AcMNPV p74 gene directly controlled the expression of SpltMNPV p74 gene in the recombinant AcMNPV bacmid-polhSL74. RT-PCR showed that the substituted p74 gene was transcribed. Bioassay showed that the recombinant virus AcMNPV bacmid-polhSL74 could not infect the Argyrogramma agnata larvae per os, and thus showing the p74 gene is species-specific.
利用大肠杆菌中RecA介导的同源重组，敲除了苜蓿银纹夜蛾多粒包埋核多角体病毒（AcMNPV）杆粒的p74基因，并用斜纹夜蛾多粒包埋核多角体病毒（SpltMNPV）的p74基因进行替代。在修饰后的p74基因座中未留下可能影响p74基因表达和功能的选择标记。AcMNPV p74基因的启动子直接控制重组AcMNPV杆粒-polhSL74中SpltMNPV p74基因的表达。RT-PCR显示替代的p74基因被转录。生物测定表明，重组病毒AcMNPV杆粒-polhSL74不能经口感染银纹夜蛾幼虫，从而表明p74基因具有种特异性。
