表达短ie-1 dsRNA的转化细胞对家蚕核型多角体病毒的抑制作用

为了利用RNAi技术提高家蚕对核型多角体病毒(BmNPV)的抗性,根据BmNPV复制必需基因ie-1设计其相应的dsRNA,构建带有ie-1 dsRNA表达盒的转基因载体piggyantiIE-Neo,结果显示:表达短ie-1 dsRNA的稳定转化Bm细胞,对BmNPV的增殖表现出抑制作用,但在病毒感染后期,由于病毒恢复增殖导致RNAi的效果被掩盖。通过反向PCR分析外源DNA片段插入基因组位点,结果显示:在转化细胞中,外源DNA可通过随机整合或按照piggyBac特定的转座位点TTAA插入细胞基因组。

Inhibitory Effects of Transformation Cells Expressing Short ie-1 dsRNA on Proliferation of Bombyx mori Nucleopolyhedrovirus

In order to confer enhanced resistance to Bombyx mori nucleopolyhedrovirus on silkworms using RNAi technology,based on ie-1,an essential gene for replication of BmNPV,the ie-1 dsRNA expression box was conceived,and thereby the transgenic vector piggyantiIE-Neo was constructed.Stable transformation Bm cells expressing short ie-1 dsRNA showed resistance to BmNPV,but during late phase of infection,the RNAi effect was concealed for the recovery of viral proliferation.Sequence analysis of the insertion site junctions of stable transformation Bm cells by inverse PCR showed that exotic DNA could be inserted into the cell genome by randomly or typically at TTAA transposition site.