In vitro expression of the HCV ARFP/F-core‘coding open reading frame

摘要

A method of producing and purifying an HCV polypeptide, wherein the method comprises providing host cells containing an expression vector comprising a polynucleotide operably linked to expression control elements, wherein the polynucleotide is an open reading frame that overlaps the core gene in the +1 frame (core+1 ORF) of HCV, which encodes an HCV polypeptide (core+1) of 16/17 kDa or 12.5 kDa is provided. The host cells can be cultured under conditions that express the HCV polypeptide (core+1) and that suppress the expression of HCV core polypeptide. The cells can be cultured in the presence of a proteosome inhibitor. An HCV polypeptide (core+1) produced by the method and an antibody that immunologically reacts with the polypeptide are provided. Isolated nucleic acids, vectors, and host cells comprising an HCV IRES element comprising core+1 sequences between nucleotides 345 and 591 are also disclosed.