新疆农业科学2017，Vol.54Issue(2)：289-296,8.DOI:10.6048/j.issn.1001-4330.2017.02.011
西瓜食酸菌TA基因酵母双杂交诱饵载体的构建及其自激活作用的检测
Construction and Self-activation Detection of Bait Vector for Yeast Two-hybrid System of TA gene in Acidovorax citrulli
摘要
Abstract
[Objective] This project aims to construct a bait vector for yeast two-hybrid system of TA gene in Acidovorax citrulli in order to lay the foundation for exploring the gene function involved in the pathogenic mechanism.[Method]The TA gene was amplified by PCR and cloned into the vector pGBKT7.The verified pGBKT7-TA was then transformed into the yeast strain AH109.The activities of report genes ADE2,HIS3 and LacZ were used to detect the self activation of pGBKT7-TA,and the toxicity of pGBKT7-TA was investigated in nutrient deficient medium.[Result]The constructed bait vector pGBKT7-TA had neither self-activation nor toxic effect in the yeast strain AH109.[Conclusion]The constructed bait vector can be used for the follow-up work of the yeast two hybrid system,which has laid a foundation for further screening of proteins interacting with TA from the cDNA library of cucumber cotyledons infected by Acidovorax citrulli.关键词
西瓜食酸菌/TA基因/酵母双杂交/诱饵载体Key words
Acidovorax citrulli/TA gene/yeast two-hybrid system/bait vector分类
农业科技引用本文复制引用
胡金凤,黄成文,刘君..西瓜食酸菌TA基因酵母双杂交诱饵载体的构建及其自激活作用的检测[J].新疆农业科学,2017,54(2):289-296,8.基金项目
国家自然科学基金项目"细菌性果斑病菌(Acidovorax citrulli)一个新型致病基因的作用机制研究"(31260419) The National Natural Science Foundation of China"Genetic and molecular characterization of a novel pathogenicity gene in Acidovorax citrulli" (31260419) （Acidovorax citrulli）
