Fusion Expression of Genes IFNα2b and Ag85B in Pichia pastoris

Ag85 complex is the mycolic acid transferase of Mycobacterium tuberculosis,and it is important in the process of forming the cell wall.Ag85B has good immunity and it is one part of Ag85 complex.Genes human IFNα2b and Ag85B were amplified by PCR,and joined by the DNA linker which encodes polypeptide (G4S)4.The recombinant fusion gene was inserted into Pichia pastoris expression vector pPIC9.The recombinant plasmid was transformed into Pichia pastoris SMD1168 and the transformants SMD1168/pPIC9-α2bAg85B were obtained.The transformants were fermented in flasks and induced by methanol.The fusion protein was identified by Western blot and was further purified.The secreted fusion protein was also identified by activity of IFNα and proved to have high activity of antivirus.