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目的构建恶性疟原虫多表位融合抗原基因(PfCP2E9),并在毕氏酵母中进行高效分泌表达。方法选取本实验室前期构建的融合蛋白PfCP-2.9与疟原虫红内期疫苗候选抗原EBA175IIF2,按一定的次序拼接成该融合基因,并克隆至酵母表达载体,用电转化方法将拼接基因导入毕氏酵母中进行分泌表达。结果拼接的基因在毕氏酵母中高水平分泌表达。结论构建的恶性疟原虫多表位融合抗原基因能在毕氏酵母中高水平分泌表达,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。
Objective To construct Plasmodium falciparum multi-epitope fusion antigen gene (PfCP2E9) and express it in Pichia pastoris efficiently. Methods The fusion protein PfCP-2.9 constructed previously in our laboratory and the candidate antigen EBA175IIF2 of erythrocytic stage of Plasmodium were selected. The fusion gene was cloned into yeast expression vector in a certain order and introduced into yeast by electrotransformation Secretion of yeast expression. Results The spliced genes were secreted at high levels in Pichia pastoris. CONCLUSION: The constructed multi-epitope antigen of Plasmodium falciparum can be secreted in Pichia pastoris at a high level, providing the basis for exploring its immunological function and as a multi-valent vaccine.