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  

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

作　　者：张鹏杰[1] 薛仁宇[1] 曹广力[1] 贡成良[1] 

机构地区：[1]苏州大学基础医学与生物科学学院,江苏苏州215123

出　　处：《蚕业科学》2008年第3期459-465,共7页ACTA SERICOLOGICA SINICA

基　　金：国家自然科学基金项目(编号30571404);教育部留学归国人员基金项目(编号K5136421);江苏省教育厅自然科学基金项目(编号05KJD230181);苏州市人才基金项目(编号2005-03);国家重点基础研究发展计划"973"项目(编号2005CB121000)

摘　　要：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。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 piggyantilE-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.

关 键 词：RNA干扰 转基因 家蚕核型多角体病毒 家蚕细胞 PIGGYBAC转座子 

分 类 号：S884.51[农业科学—特种经济动物饲养] Q78[农业科学—畜牧兽医]