恶性疟原虫红前期多表位融合抗原基因的构建及其高效表达

目的:构建一个恶性疟原虫红前期多表位融合抗原基因(PfCP-TCL),并在毕氏酵母中进行高效分泌表达.方法:选取了疟原虫红外期疫苗候选抗原CSP、TRAP和LSA-1中的一些重要T、B细胞表位,按一定的次序加以串联连接,并全合成该融合基因.在毕氏酵母中进行分泌表达,并纯化表达产物.结果:合成的基因在毕氏酵母中高水平分泌表达,产量达1 g/L以上.经离子交换和凝胶过滤2步纯化过程,获得纯度在95%以上的重组蛋白.结论:全合成的多表位融合抗原基因能在毕氏酵母中高水平分泌表达,并产生一种工艺简易的纯化方法.大量获得该重组蛋白为探讨其免疫学功能及作为多价联合疫苗的成分提供了基础.

Constructing a pre-erythrocytic multi-epitope chimeric antigen of Plasmodium falciparum and its expression in Pichia pastoris

Objective:To construct a pre erythrocytic mult i epitope antigen of Plasmodium falciparum (designated as PfCP TCL ) and express it in Pichia pastoris in secreting form. Methods: Some importa nt T and B epitopes from pre erythrocytic antigen candidates such as CSP, TRAP a nd LSA 1 were selected and joined in tandem to generate the chimeric gene. The gene was expressed in Pichia pastoris and its product was purified by chroma togr aphy. Results: PfCP TCL gene was synthesized and expressed at h igh level of more than 1 g/L. The product was purified by ion exchange and gel filter chromatography with a purity of >95%. Conclusi on : The synthetic PfCP TCL gene is expressed in Pichia pastoris and a method of easy purification is developed. The recombinant PfCP TCL protein provides a base for investigating its immune function and potential as a component of combined malaria vaccine.