恶性疟原虫膜蛋白Pfs25在毕赤酵母中的组成型表达

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BstXⅠ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/GS115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。

Constitutive Expression of Plasmodium falciparum Transmission-Blocking Vaccine Antigen Pfs25 in Pichia pastoris

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 pGAPZαA to construct recombinant plasmid pfs25/pGAPZαA,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.