中文摘要：

家蚕核型多角体病毒（Bombyx mori nucleopolyhedrovirus,BmNPV）是影响蚕业生产最为严重的病原之一,每年对我国蚕业生产方面造成很大的经济损失。通过RNA干扰技术（RNAi）干扰BmNPV的增殖关键基因可以有效地抑制病毒的增殖复制。本研究以BmNPV侵染关键基因gp64和lef-1为靶标基因,并利用病毒诱导型启动子LEF3P和39KP共构建了4种RNAi干扰表达载体,分别命名为：piggyBacA3-EGFP-39KP-gp64、piggyBacA3-EGFP-39KP-lef-1、piggyBacA3-EGFP-LEF3P-gp64、piggyBacA3-EGFP-LEF3P-lef-1。经瞬时转染和筛选稳定表达的家蚕细胞系抗病毒检测结果表明,通过RNAi技术能够有效的抑制病毒增殖复制,并确定了39KP启动效果优于LEF3P启动子,干扰gp64基因的抗病毒效果优于lef-1基因。这些研究结果为后期转基因品系培育和家蚕抗病毒研究提供了基础。

英文摘要：

Bombyx mori nucleopolyhedrovirus（BmNPV）is causes huge economic losses to the sericulture production in China each year.RNA interference（RNAi）of the key genes of BmNPV can inhibit the proliferation and replication of BmNPV.We constructed four RNAi expression vectors（piggyBacA3-EGFP-39KP-gp64,piggyBacA3-EGFP-LEF3P-lef-1,piggyBacA3-EGFP-LEF3P-gp64 and piggyBacA3-EGFPLEF3P-lef-1）using the inducible promoters LEF3 Pand 39KP promoter to initiate the target genes of gp64 and lef-1.Use of silkworm cell lines showed that RNAi could effectively inhibit BmNPV replication,and that the effect of the 39 KP promoter was better than that of the LEF3 Ppromoter.Furthermore,the interference effect of gp64 gene was better than that of lef-1gene.These results provide a basis for the development of transgenic lines and the study of antiviral activity in silkworms.