中国兽医科学2011，Vol.41Issue(5)：470-473,4.
鸭肠炎病毒感染鸭肝酵母双杂交cDNA文库的构建
Construction of yeast two-hybrid cDNA library of duck's liver inoculated with duck enteritis virus
摘要
Abstract
To construct the yeast two-hybrid cDNA library of duck's liver infected with duck enteritis virus(DEV) ,total RNA was isolated from duck's liver infected with DEV by the Trizol method.The first and second strand cDNA were synthesized by reverse transcriptase and DNA polymerase Ⅰ , respectively.The amplified cDNA was cloned into the pGADT7 yeast expression vector and over-expressed in Y187 strain.The non-diluted,1 : 10 and 1: 100 dilution inoculated on SD/-Leu for 3 days at 30 ℃ ,respectively.Total size of the library transformed into Y187 strain was 1 × 106 clones.The insert size by PCR from the colony and plasmids from the library was from 0.5 to 1 kb.This yeast two-hybrid library had good quality and facilitated the screening duck's liver proteins interacting with DEV NP from cDNA library.关键词
酵母双杂交/鸭肠炎病毒/cDNA文库Key words
yeast two-hybrid system/duck enteritis virus/cDNA library分类
农业科技引用本文复制引用
张荷,毛君婷,杨颖,王开功,周碧君,文明..鸭肠炎病毒感染鸭肝酵母双杂交cDNA文库的构建[J].中国兽医科学,2011,41(5):470-473,4.基金项目
国家自然科学基金资助项目(30760182) （30760182）
