Screening of interferon-β binding proteins by yeast-two hybrid technique from a cDNA library

Objective To screen binding proteins of interferon β(IFNβ)from human hepatic cDNA libraty by yeast-two hybrid technique.Methods The IFNβ gene was amplified by polymerase chain reaction(PCR)and pGBKT7-IFNβ plasmid was constructed with molecular biology technique.pGBKT7-IFNβ was transfected into yeast AH109.The transfected yeast was mated with yeast Y187 containing liver cDNA library plasmid in 2×YPDA medium.Diploid yeast was plated on synthetic dropout nutrient medium(SD/-Trp-Leu-His-Ade)and synthetic dropout nutrient medium(SD/-Trp-Leu-His-Ade)containing X-α-gal for selecting.After plasmid extracting and enzyme cutting analysis,sequence analysis was performed on the blue colonies,the results were analyzed by bioinformatics.Results Nineteen genes in twenty-nine positive colonies were obtained,coding for 15 kinds of proteins,including 1 protein with unknown function.Conclusion IFNβ binding proteins were screened using yeast-two hybrid technique.