美国白蛾中肠cDNA表达文库的构建与鉴定  

Construction and identifi cation of cDNA expression library from Hyphantria cunea larvae midgut

作　　者：刘海明[1] 宋雯雯[1] 谭秀梅[1] 郑桂玲[1] 李长友[1] 周洪旭[1] 

机构地区：[1]青岛农业大学农学植物保护学院,山东青岛266109

出　　处：《植物检疫》2014年第1期25-28,共4页Plant Quarantine

基　　金：山东省教育厅项目(J10LC07);青岛农业大学博士基金(6631024);‘泰山学者’建设工程专项经费

摘　　要：美国白蛾是一种重要的外来入侵性害虫，其围食膜是害虫生物防治的潜在靶标。本研究以美国白蛾中肠为材料，通过提取其总RNA、分离纯化mRNA、反转录合成cDNA等方法，构建了美国白蛾中肠cDNA表达文库。测得文库的原始滴度为4．08×10^5pfu／mL，蓝白斑鉴定该文库的重组率为95．7％，插入片段大小在0．8~2．3kb之间，平均长度为1．77kb。将所得到原始文库扩增后进行保存，检测扩增文库滴度为3．92×10^8pfu／mL。美国白蛾中肠cDNA表达文库的成功构建为克隆美国白蛾围食膜蛋白基因，开发环保型微生物杀虫剂提供可靠的理论依据。Hyphantria cunea （Drury） is an important invasive pest, and the peritrophic matrix is a potential target for biological control of pests. In this paper, the eDNA expression library from Hyphantria cunea larvae midgut was constructed by total RNA extraction, mRNA purification and reverse transcription eDNA synthesis methods. The primary library titer of 4.08~ 105pfu/mL was obtained and the recombination efficiency was 95.7%. The length of inserted fragments varied from 0.8 to 2.3kb with the average length of 1.77kb. The eDNA library was amplified and stocked, the titer of which was estimated as 3.92×108pfu/mL. This high quality eDNA library provided a reliable theoretical basis for cloning peritrophie matrix protein gene from Hyphantria cunea larvae and developing environment-friendly microbial insecticides.

关 键 词：美国白蛾 围食膜 CDNA文库 基因克隆 

分 类 号：S433.4[农业科学—农业昆虫与害虫防治]