Construction and identification of bait vector pGBKT7-COPS3 for screening COPS3 interaction protein

Objective To construct bait vector pGBKT7-COPS3 in yeast two-hybrid system GAL4,and explore whether expression product of COPS3 has toxicity to host yeast cells and activates the reporter genes in the yeast two-hybrid system.Methods The COPS3 gene fragments were amplified by RT-PCR from SOSP-9607,and cloned into pUC19 for DNA sequencing.After verified by sequencing,it was subcloned into the bait vector pGBKT7 of yeast two-hybrid system GAL4,and the recombinant plasmid was transferred into yeast cell AH109.The activation of expression product to the reporter genes was tested in the yeast two-hybrid system.Results The COPS3 gene fragments were successfully amplified,and the expression product had no toxicity to AH109 cells,and could not activate the reporter genes of the GAL4 system.Conclusion Yeast two-hybrid GAL4 system can be utilized to study COPS3 interaction protein.