Production of the Major Merozoite Surface Protein 1(MSP1) of Plasmodium falciparum in Pichia pastoris
恶性疟裂殖子表面蛋白1合成基因在毕赤酵母中的表达

The major Merozoite Surface Protein 1(MSP1) of Plasmodium falciparum is an important candidate for malaria vaccine.Highly instability of msp1 gene due to its unusual high AT content (74%),however,render cloning of the full\|length of this gene impossible.Synthesis of the entire gene using other codon usage has provided possibility to produce the entire MSP1 in heterogeneous system.In this investigation,the entire MSP1 recombinant protein has been expressed in Pichia pastoris. Insertion of the synthetic msp1 gene into the pichia expression vector pPIC3 5 generated the recombinant plasmid pPIC3 5/msp1 and transformation of the Pichia pastoris SMD1168 by electroporation produced recombinant transformants which were verified by PCR amplification of msp1\|derived fragment from the chromosome.The recombinant MSP1 derived from Pichia was interacted strongly with monoclonal antibody mAb5 2 recognizing the disulfide\|bond\|depended conformational epitope at the C\|terminus of MSP1 molecule,indicating that the protein generated in this system resembled most likely to native protein at least at this conformational epitope.Availability of isolating the recombinant protein from the yeast provides a possibility to examine the protective potential of the molecule as a vaccine candidate.