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 fra...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.展开更多
文摘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.
