Dongmei Zhang1, Weiqing Pan, Deru Lu, Liping Jiang. 1. Institute of Medical Biotechnology & Molecular Genetics of Second Military Medical University, Shanghai 200433 China.
Abstract
OBJECTIVE: Production of 3D7/MSP1 - 42 recombinant protein with correct conformation in Pichia pastoris for vaccine efficiency assay. METHODS: Asymmetric PCR-based method was utilized to synthesize the 1 202 bp 3D7/msp1 - 42 gene. The expressing plasmid containing the synthetic gene was introduced into Pichia pastoris by electroporation. The secreted product was detected by Western Blot. RESULTS: The redesigned entire 3D7/msp1 - 42 gene was generated with error-free, and expressed to produce 42 kD recombinant protein in secreted form. Conformational monoclonal antibody specific for MSP1 C-terminal can interact with the recombinant protein. CONCLUSION: The redesigned 3D7/msp1 - 42 gene was expressed in P. pastoris with full length of recombinant protein which resembled most likely to the native protein.
OBJECTIVE: Production of 3D7/MSP1 - 42 recombinant protein with correct conformation in Pichia pastoris for vaccine efficiency assay. METHODS: Asymmetric PCR-based method was utilized to synthesize the 1 202 bp 3D7/msp1 - 42 gene. The expressing plasmid containing the synthetic gene was introduced into Pichia pastoris by electroporation. The secreted product was detected by Western Blot. RESULTS: The redesigned entire 3D7/msp1 - 42 gene was generated with error-free, and expressed to produce 42 kD recombinant protein in secreted form. Conformational monoclonal antibody specific for MSP1 C-terminal can interact with the recombinant protein. CONCLUSION: The redesigned 3D7/msp1 - 42 gene was expressed in P. pastoris with full length of recombinant protein which resembled most likely to the native protein.