人Era不同结构域在酵母双杂交系统中的表达及对报告基因激活作用的检测  

Reporter Genes Assay of the Different Domain of Human Era in Yeast Two-hybrid System

作　　者：吴元明[1] 程轶梦[1] 陈苏民[1] 陈南春[1] 纪宗玲[1] 

机构地区：[1]第四军医大学生物化学与分子生物学教研室,西安710032

出　　处：《中国生物化学与分子生物学报》2002年第2期254-258,共5页Chinese Journal of Biochemistry and Molecular Biology

基　　金：国家自然科学基金资助项目 (No .39870 380 ;396 70 0 0 6 )~~

摘　　要：To construct the bait vector with different domains of human Era in yeast two-hybrid system, then assay whether its expression product can affect the growth of yeast cells and activate the reporter genes, the cDNA fragments encoding different domains of human Era was amplified by PCR, and subsequently they were cloned into pUC19. After being verified by sequencing, they were subcloned into the bait vector pGBKT7 of yeast two-hybrid system. Furthermore, these recombinant plasmids were transferred into AH109, and their expression products were assayed whether they could activate the reporter genes. The cDNA fragments encoding different domains of human Era were amplified successfully. The different domains of human Era were not toxic to AH109 and could not activate the reporter genes. Yeast two-hybrid GAL4 system could be used to fish human Era interacting protein.To construct the bait vector with different domains of human Era in yeast two-hybrid system, then assay whether its expression product can affect the growth of yeast cells and activate the reporter genes, the cDNA fragments encoding different domains of human Era was amplified by PCR, and subsequently they were cloned into pUC19. After being verified by sequencing, they were subcloned into the bait vector pGBKT7 of yeast two-hybrid system. Furthermore, these recombinant plasmids were transferred into AH109, and their expression products were assayed whether they could activate the reporter genes. The cDNA fragments encoding different domains of human Era were amplified successfully. The different domains of human Era were not toxic to AH109 and could not activate the reporter genes. Yeast two-hybrid GAL4 system could be used to fish human Era interacting protein.

关 键 词：人ERA 结构域 酵母双杂交系统 表达 报告基因 激活作用 检测 

分 类 号：Q75[生物学—分子生物学] R394.8[医药卫生—医学遗传学]