Overexpression of an epitope of HCV displayed in a new foreign epitope presenting system / 中华传染病杂志

ABSTRACT

Objective An epitope in E1 region of hepatitis C virus (HCV) was constructed and highly expressed in a foreign epitope presenting vector based on an insect virus, flock house virus capsule protein encoding gene as a vector. Methods Using genetic engineering techniques, an epitope in E1 region of HCV was constructed in a foreign epitope presenting vector based on an insect virus, flock house virus capsule protein encoding gene as a vector and lastly constructed a chimeric gene. The chimeric gene can be put into an expressing plasmid, and expressed in either eukaryotic or prokaryotic system. The foreign epitope can be expressed in this system on the surface of the vector stereotic structure with its native conformation, and which can improve its immunogenecity. Results An epitope in E1 region of HCV was successfully cloned in three positions (aa106、aa153、aa305) of a foreign epitope presenting system based on an insect virus, flock house virus capsule protein encoding gene as a vector.The recombinant epitope in this system could be highly expressed more than 40% of total cell protein in E.coli.BL21 cell in somewhat strict conditions without IPTG inducing. Conclusions We have made this chimeric gene highly expressed in E.coli.BL21, by the way different from routine method. The results suggest that the expression method and this epitope presenting system can be used in studying immunological and biological properties of HCV and developing diagnostic reagents as well as epitope-based subunit vaccines for controlling HCV infection.