Screening of proteins binding to hepatitis C virus E1 protein from human pancreas cDNA library / 中华传染病杂志

ABSTRACT

Objective To screen proteins from human pancreas cDNA library,which interact with hepatitis C virus(HCV)E1 protein.Methods The human pancreas cDNA library was amplified,purified and evaluated,and then the purified library plasmids were transformed into yeast strain Y187.The reconstructed plasmid pGBKT7-E1 was transformed into yeast strain AH109 and screened on the nutrient deficiency medium SD/-Trp.The transformed AH109 mated with Y187 that contained the library plasmids.The diploid yeast cells were plated on nutrient deficiency medium SD/-Trp/-Leu/-His/-Ade and SD/-Trp/-Leu/-His/-Ade containing X-α-gal for selecting.The plasmids in diploid yeast cells were extracted and electrotransformed into E.coli DH5α.The plasmids in DH5α were extracted,sequenced and blasted.Result Sixteen proteins interacting with HCV E1 were found.Conclusion Some of the sixteen pancreatic proteins may be related with metabolisms of glucose and lipid.