Expression and immunogenicity of recombinant Annexin32 in methylotrophic yeast Pichia pastoris

Objective:To express and identify Annexin32 in meythylotrophic yeast Pichia pastoris .Methods:Nine high copy number transformants of Pichia pastoris ,whose phenotypes were screened by culture and PCR,were induced by methanol in BMMY,BMM and MM for the expression of Annexin32.Supernatants after induction were analyzed by SDS PAGE and Western blot.The Annexin32 were separated and purified by (NH 4) 2SO 4+PI and Superdex75.The pure Annexin32 were then immunized to mice.Results:Two Mut s transformants gave high level expression in BMMY.The expression product had 2 bands (MW 38 000 and 40 000) and the concentration was 200 350 mg/L,accounting for over 80% of the total secreted protein.Our data showed higher immunogenicity in Annexin32 from yeast than from E.coli .Conclusion:Annexin32 is expressed successfully in mythylotrophic yeast Pichia pastoris ,providing a foundation for further research.