选择家蚕核型多角体病毒(Bm NPV)基因组中可能与病毒增殖相关的8个基因dbp、lef-2、lef-3、lef-7、lef-10、lef-11、p143和vp1054作为候选基因,利用家蚕内源性microRNA骨架构建候选基因的干扰载体,并采用RNA干扰(RNA interference,RNAi)技术对上述候选基因进行干扰试验,筛选病毒增殖的关键基因,以期为开辟家蚕抗Bm NPV研究新路径打下基础。对8个候选基因的RNA干扰试验显示:干扰lef-7对Bm NPV的增殖没有明显影响;对lef-2和lef-11表达的干扰效率最高,达到90%以上,其中干扰lef-11后病毒对细胞的感染率仅为9.16%,远远低于对其他基因干扰后的感染率。依据试验结果初步确认8个病毒增殖相关基因中,lef-11对病毒增殖的影响最大。
In this study,eight proliferation-related genes dbp,lef-2,lef-3,lef-7,lef-10,lef-11,p143 and vp1054 in Bombyx mori nucleopolyhedrovirus( Bm NPV) were selected as candidate genes. We used silkworm( Bombyx mori) endogenous microRNA skeleton to construct corresponding interference vectors to silence these candidate genes by RNA interference( RNAi) technology,so as to identify the key proliferation-related genes and provide a good basis for novel anti-BmNPV research in silkworm. RNA interference test showed that interfering lef-7 gene had no obvious effect on Bm NPV proliferation,and interfering lef-2 and lef-11 genes had the highest efficiency( over 90%). After interfering lef-11 gene,infection rate of the virus was reduced to only 9. 16%,which was much lower than that of interfering other genes. These results demonstrate that lef-11 is the pivotal gene among the eight genes related to proliferation and replication of Bm NPV.