表达短lef-1 dsRNA转基因家蚕对BmNPV的抵抗能力与主要经济性状  

Resistance to BmNPV and Main Economical Traits of Transgenic Silkworms Expressing Short lef-1 dsRNA

作　　者：叶秋霞[1] 薛仁宇[1] 曹广力[1] 张鹏杰[1] 潘中华[1] 郑小坚[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》2011年第5期825-831,共7页ACTA SERICOLOGICA SINICA

基　　金：国家自然科学基金项目(No.31072085;30571404);国家重点基础研究发展计划"973"项目(No.2005CB121005);苏州大学重大应用研究培育项目(No.Q3134991)

摘　　要：为了提高家蚕对核型多角体病毒(BmNPV)的抵抗能力,将带有BmNPV晚期表达因子基因(lef-1)dsRNA表达盒和新霉素抗性基因(neo)表达盒的转基因载体pigA3-LEF-Neo通过精子介导法导入家蚕卵,经G418和GFP双重筛选并结合分子生物学鉴定,证实获得了转基因家蚕。对G6代转基因家蚕2龄幼虫接种BmNPV,结果显示转基因家蚕的死亡率比正常家蚕(对照组)下降了10～30个百分点,表明转基因家蚕对BmNPV的抗性有所提高;对接种BmNPV的5龄转基因家蚕A、B系统的定量PCR检测结果显示,2个系统体内的lef-1 mRNA转录水平比对照组平均降低了72倍和26.5倍,最高下降了104倍,进一步表明表达lef-1 dsRNA的转基因家蚕抑制了BmNPV lef-1基因的表达。转基因家蚕的饲养成绩显示其全茧量、茧层量和茧层率等主要经济性状与正常家蚕无明显差异。In order to improve the resistance of silkworm（Bombyx mori） to nucleopolyhedrovirus（BmNPV）,the transgenic vector pigA3-LEF-Neo containing BmNPV＇s late expression factor 1（lef-1） dsRNA and neomycin resistance gene（neo） expression cassette was transferred into the eggs of silkworm by sperm-mediated gene transferring method.Screening with GFP and antibiotics G418 in combination with molecular biological identification verified that transgenic silkworms were generated.The mortality of G6-generation 2nd instar transgenic silkworms infected with BmNPV decreased by 10 to 30 percentage points compared with normal ones（control group）,demonstrating that the resistance of transgenic silkworms to BmNPV has been improved.Quantitative PCR assay revealed that,after inoculation with BmNPV,the average lef-1 mRNA transcriptional levels in the 5th instar larvae of transgenic silkworm lineages A and B declined by 72 folds and 26.5 folds with the highest as 104 folds compared to the control group.These results further confirmed that the transcribed lef-1 dsRNA in transgenic silkworms had inhibited the expression of the lef-1 gene.Statistics on economic traits showed that there were no significant differences in the whole cocoon weight,cocoon shell weight and cocoon shell percentage between transgenic silkworms and normal silkworms.

关 键 词：家蚕核型多角体病毒 晚期表达因子基因 双链RNA 转基因家蚕 抗性 经济性状 

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]