Identification of HCV core protein binding proteins by yeast two-hybrid

Abstract

To identify HCV core protein binding proteins. The library was amplified, purified, and then were transformed into yeast strain Y187. The reconstructed plasmid pGBKT7-core was transformed into yeast strain AH109 and screened on the nutrient deficiency medium SD/-Trp. The transformed AH109 mated with Y187 containing the library plasmid. The diploid yeast cells were plated on nutrient deficiency medium SD/-Trp/-Leu/-His/-Ade and SD/-Trp/-Leu/-His/-Ade containing X-alpha-gal for selecting. The plasmids in diploid yeast cells were extracted and electrotransformed into E.coli DH5alpha. The plasmids in DH5alpha were extracted and sequenced. Eleven proteins, including chymotrypsinogen B1 precursor, carboxypeptidase A1, trypsinogen 2, chymotryptic peptide C, trypsin 1, carboxypeptidase B1, kinesin superfamily proteins 3B, trypsin 2, mitochondria protein gene, elastase 3A and colipase were found to be able to bind to HCV core protein. Proteins related with metabolism of glucose and lipid may bind to HCV core protein.