Construction and Identification of Bait Vectors with VP2 and VP3 Gene of Goose Parvovirus in Yeast Two-Hybrid System

Two bait vectors,pGBKT7-VP2 and pGBKT7-VP3,were constructed for screening cellular proteins interacting with VP2 and VP3 proteins of goose parvovirus from yeast two-hybrid cDNA library of goose embryo fibroblast cells in this study. VP2 and VP3 genes were amplified by PCR from plasmid pGEX-4T-VP1 and cloned into pMD-18T vector. After being verified by sequencing,they were subcloned into vector pGBKT7 of yeast two-hybrid system. Then the recombinant plasmids identified by PCR,enzyme digestion and sequencing were transformed into yeast cells H2YGold. The two bait vectors' self-activation to reporter genes and cytotoxicity to the yeast cells were tested. The results showed that the two bait plasmids,pGBKT7-VP2 and pGBKT7-VP3,were successfully constructed and proved to be no self-activation to reporter genes and no cytotoxicity to the yeast cells.They could be used to screen cellular proteins interacting with VP2 and VP3 proteins in the yeast two-hybrid system.