Cloning and expression of ns5a gene of hepatitis C virus 1b strain DY in Escherichia coli / 西安交通大学学报(医学版)

ABSTRACT

Objective To clone and express the ns5a gene of hepatitis C virus(HCV) 1b strain DY.Methods By using the prokaryotic cell gene engineering,HCV ns5a gene was amplified with nested PCR from the plasmid HCV17 of HCV 1b strain DY full-length gene and inserted into the cloning pMD18-T vector.The cloned HCV ns5a gene was separated and subcloned into expression vector pET-28a and induced by IPTG in E.coli.BL21.The expressed product was identified by SDS-PAGE and Western-blot methods.Results Recombinant expression plasmid pet-28a-ns5a was constructed and expressed successfully.Conclusion HCV ns5a gene was cloned and expressed.This might be helpful for further studies on the nature and biological properties of the ns5a gene.