大草蛉Chrysopa pallens(Rambur)触角cDNA文库的构建  

Construction of cDNA library of Chrysopa pallens (Rambur) antenna

作　　者：王娟[1] 张礼生[1] 张海平[1] 刘晨曦[1] 王孟卿[1] 陈红印[1] 

机构地区：[1]中国农业科学院植物保护研究所农业部作物有害生物综合治理重点实验室、中国-美国生物防治实验室,北京100081

出　　处：《环境昆虫学报》2014年第6期898-904,共7页Journal of Environmental Entomology

基　　金：“973”计划(2013CB127602);公益性行业(农业)科研专项(201103002);“948”项目(2011-G4)

摘　　要：本文以羽化3天内的大草蛉Chrysopa pallens(Rambur)雌雄触角为材料提取总RNA,以此为模板,通过SMARTscribeTM Reverse Transcriptase反转录酶反转录合成第一链cDNA,引物扩增获得双链cDNA,经CHROMA SPIN+TE-1000 Column分级分离后,收集0.4-2.0 kb之间的片段重组于质粒载体pSMART2IFD并电转化至大肠杆菌Escherichia coli DH5ɑ,最终构建获得大草蛉雌雄触角全长cDNA文库。结果显示构建的雌雄触角cDNA初级文库滴度分别2.1×106pfu/mL和1.8×106pfu/m L,重组率均达到93.0%以上,插入cDNA的片段大小范围为0.4-2.0 kb,主要集中在0.5-1.0 kb,平均长度为549.0 bp,表明构建获得的文库质量较高,可保证大规模全长c NA的获得。该文库的构建丰富了大草蛉基因序列信息,为大量克隆大草蛉嗅觉相关基因,深入揭示大草蛉嗅觉识别机制奠定基础,也为合理利用大草蛉进行生物防治提供理论依据。A full - length cDNA library from the antenna of Chrysopa pallens （Rambur） was constructed based on switching mechanism at 5′end of RNA transcript （SMART） system. The purified double strand cDNA was ligated to vector SMART2IFD, and transformed into Escherichia coli DH5α by electroporation. The library constructed for female and male have high titer of 2. 1 × 10^6 pfu/mL and 1.8 × 10^6 pfu/mL, respectively. PCR results showed that the inserts varied from 400 bp to 2000 bp with average size larger than 500 bp and 93.0% of recombinant percentage. Further studies on those genes and large scale sequencing of the library may be helpful to screen newly olfaction relative genes in C. pallens.

关 键 词：大草蛉 触角 嗅觉 CDNA文库 

分 类 号：Q965[生物学—昆虫学] S476[农业科学—农业昆虫与害虫防治]