Objective: Malaria is the mosquito-borne infectious disease and Pfs25 protein is the antigen for transmission-blocking vaccine. The aim of this study is to express the Pfs25 protein in Pichia pastoris. Methods: According to the codon-like principle for P.pastoris, the target gene was synthesized and inserted into a yeast constitutive vector pGAPZaA to construct recombinant plasmid pfs25/pGAPZaA, and then transformed the host P.pastoris GS115 genome by electroporation. The expressed product was identified by SDS-PAGE and Western blot. The recombinant P.pastoris was analyzed for genetic stability. Results: The Pfs25 protein was expressed successfully in P. Pastoris, and the yeast transformants still had the foreign gene after eight passages in YPD medium. Conclusion: This study laid a foundation of future research in malaria transmission-blocking vaccine.%目的:Pfs25蛋白是传播阻断型恶性疟疾疫苗的侯选抗原,在毕赤酵母中表达Pfs25蛋白,并对表达产物进行鉴定.方法:参照GenBank中公布的pfs25基因序列,通过毕赤酵母喜好密码子分析人工合成目的基因；采取定向克隆策略构建重组表达质粒pfs25/pGAPZαA,经BstX I线性化,电转染法转化酵母菌株GS115,在Zeocin抗性的筛选培养基上获得表达目的基因的pfs25/pGAPZαA/GS 115重组酵母菌,SDS-PAGE和Western印迹检测表达产物；通过在YPD培养基上传代培养和目的基因表达,验证重组菌株的遗传稳定性.结果:在毕赤酵母中表达了Pfs25蛋白,且重组菌株遗传性质稳定.结论:为研制基于Pfs25蛋白的传播阻断型恶性疟疾疫苗奠定了基础.
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