Related Experiment Video
Updated: Aug 6, 2026

A Protocol for Analyzing Hepatitis C Virus Replication
Published on: June 26, 2014
Preparation of hepatitis C virus structural and non-structural protein fragments and studies of their immunogenicity
Marija Mihailova1, Melanie Fiedler, Mechthild Boos
1Institute of Virology, Essen University Hospital, Hufelandstr. 55, 45122 Essen, Germany. mary@biomed.lu.lv
Abstract:
Plasmids pQE-60 and pQE-30 containing 6 x His-tag sequence were used for expression of fragments of HCV structural and non-structural proteins in Escherichia coli (E. coli). The following fragments were used: core (1-98 aa), NS3 (202-482 aa), and tetramer of hypervariable region 1 (HVR1) of E2 protein. The constructed plasmids directed high levels of expression of HCV proteins in E. coli JM109. After purification by the metal-affinity chromatography on nickel-nitrilotriacetic acid (Ni-NTA) agarose, the His-tagged HCV proteins were used for immunization of BALB/c mice. All three proteins were able to induce high levels of specific antibodies and, in the case of the NS3 and HVR1 tetramer, also to mount vigorous cell-proliferating responses. High immunogenicity of the tested fragments of HCV proteins shows them as good candidates for inclusion into the future HCV vaccine preparations.

