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选用苜蓿丫纹夜蛾核多角体病毒杆粒 (AcMNPVbacmid)为材料 ,通过在大肠杆菌中利用RecA基因介导的同源重组 ,将其p74基因剔除 ,并精确地用斜纹夜蛾核多角体病毒 (SpltMNPV)的p74基因进行了替换。所构建的重组AcMNPV杆粒在修饰后的p74基因位点中未留下任何有可能影响该基因表达及功能的选择标记 ,SpltMNPV的p74基因直接位于AcMNPVp74基因的启动子控制下。RT PCR显示替换后的p74基因得到了表达。生物测定结果显示 ,重组病毒AcMNPV杆粒 polhSL74无法通过口服方式感染银纹夜蛾幼虫 ,表明杆状病毒p74基因具有种属特异性。
In this study, AcMNPVbacmid was used as a material. The p74 gene was deleted by homologous recombination mediated by RecA gene in Escherichia coli. The p74 gene was deleted and amplified with Spodoptera litura nucleopolyhedrovirus (SpltMNPV) p74 gene was replaced. The constructed recombinant AcMNPV bacmid did not leave any selectable marker in the modified p74 gene site that may affect the expression and function of the gene. The p74 gene of SpltMNPV is directly under the control of the promoter of AcMNPVp74 gene. RT PCR showed that the replaced p74 gene was expressed. Bioassay results showed that the recombinant virus AcMNPV baculovirus polhSL74 could not orally infect S. litura larvae, indicating that the baculovirus p74 gene is species-specific.