摘要
以草地螟中肠为材料,分别采用、Oligotex mRNA kit、ZAP-cDNA synthesis kit和ZAP-cDNA GigapackⅢGold Cloning Kit提取草地螟幼虫中肠总RNA,分离纯化mRNA并构建了草地螟中肠cDNA表达文库。测得文库的原始滴度为3.73×10~6pfu/ml,蓝白斑测定重组率达99.7%,插入片段平均长度约为1.8 kb。草地螟中肠cDNA表达文库的成功构建为利用文库筛选中肠靶标蛋白提供了材料来源,为研制、开发草地螟生防药剂的新靶标以及研究其作用的分子机制奠定了基础。
Midgut epithelial tissues of Loxostege sticticalis was was used as experimental materials and Loxostege sticticalis midgut cDNA library was constructed.First,total RNA was extracted and mRNA was purifed by using RNeasy mini kit and Oligotex mRNA kit,then according to ZAP-cDNA~ synthesis kit and ZAP-cDNA~ Gigapack~ⅢGold Cloning Kit constructed the midgut cDNA expression library.A primary library titre of 3.73×10~6Pfu/mL was obtained.The recombinant efficiency was 99.7%.The average length of fragment inserted was 1.7 kb.This high quality cDNA library can be further used for screening target protein of midgut and provides useful tool for the study of the molecular mechanisms of its functional genes that may be useful for designing novel pest control methods.
出处
《生物技术通报》
CAS
CSCD
北大核心
2009年第S1期219-223,共5页
Biotechnology Bulletin
基金
国家科技支撑课题(2005BA529A03)
关键词
草地螟
中肠
CDNA表达文库
Loxostege sticticalis
Midgut
cDNA expression library
