医学分子生物学杂志Issue(3)：139-143,5.DOI:10.3870/j.issna.1672-8009.2014.03.003
人星状病毒非结构蛋白酵母双杂交诱饵表达载体构建及酵母转化子特性鉴定
Construction of Yeast Two-Hybrid Bait Vector of HAstV Non-Struc-tural Protein and Characterization of Yeast Transformants
摘要
Abstract
Objective To construct a yeast two-hybrid bait vector of non-structure protein 1a/3 (nsP1a/3) of human astrovirus (HAstV), and to characterize the yeast transformants after elec-trotransformation of the recombinant plasmid into the yeast competent cells .Methods The coding sequence of HAstV nsP 1 a/3 obtained by PCR was inserted into the modified plasmid pGST 7.The re-combinant vector nsP1a/3-pGBST7 confirmed by DNA sequencing was transformed into the Y 2 HGo-ld yeast competent cells .The cytotoxicity of the expression products of the recombiant plasmid nsP1a/3-pGBST7 and the autoactivation of the report gene in yeast transformants were exam-ined.Results The sequencing of the nsP 1 a/3-pGBST7 vector showed that the recombinant bait plasmid was successfully constructed and the open reading frame of nsP 1a/3 insert was correct.After electrotransformation, it was found that the expression products of the nsP 1a/3-pGBST7 vector had no cytotoxicity to yeast transformants and also they did not activate the report gene .Conclusion The yeast two-hybrid bait vector of HAstV nsP 1 a/3 was successfully constructed , which laid a foun-dation for further studying the interaction of HAstV nsP 1 a/3 with target proteins and the replication mechanism of HAstV in host cells .关键词
人星状病毒/nsP1a/3基因/酵母双杂交/转化子特性Key words
human astrovirus/nsP1a/3 gene/yeast two-hybrid/transformant characteristics分类
医药卫生引用本文复制引用
李攀,崔成成,杨思达,黄芬,曾韦锟,井申荣..人星状病毒非结构蛋白酵母双杂交诱饵表达载体构建及酵母转化子特性鉴定[J].医学分子生物学杂志,2014,(3):139-143,5.基金项目
国家自然科学基金（No．31360619，31160193），云南省应用基础研究面上项目（No．2010ZC055，2012FB135）This work was supported by grants from the National Natural Science Found-aion of China （No．31360619，31160193） and Based on Application on the Project of Yunnan Province （）
