Summary
Cloning and Expression of Therapeutic Recombinant Interferon Gamma
Interferon gamma (IFN-γ), the unique member of class II interferon, plays a very important role in treatment of viral infection and liver fibrosis. In this study, the expression vector pNanogenIFN-γ anchoring gen encoding for IFN-γ was cloned and transformed onto E.coli BL21 (DE3) bacterial strain to make the E.coli BL21 (DE3) pNanogenIFN-γ strain. This strain can proliferate and be induced to produce IFN-γ in LBkan after 5.5h cultivation at 37oC with the induction condition of 0.5mM IPTG, induction time of 6h. Purified protein using FPLC purification process including re-folding, buffer exchange, cation exchange chromatography and gel filtration was > 95% purity and the recovery rate of the purification was 24.5%.
Keywords: Cloning, interferon gamma, inclusion body, over expression, re-folding, protein purification.
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