【摘要】 通过家蚕杆状病毒表面展示技术获得约氏疟原虫有性阶段候选抗原Pys25(217AA)和Pys48(455AA),以期建立一种新的鼠疟模型。首先利用大肠杆菌原核表达系统表达这两种抗原,免疫Balb/c小鼠制备多克隆抗体,同时利用家蚕杆状病毒表面展示技术,并改造pFastBac Dual载体,在其Ph启动子前端加入哺乳动物启动子CMV,并将目的蛋白基因与杆状病毒囊膜蛋白gp64基因片段融合表达,融合蛋白展示在病毒粒子表面。用其免疫Balb/c小鼠,在小鼠体内,CMV启动子启动融合蛋白表达,共同刺激小鼠产生多克隆抗体。通过间接ELISA检测,两种抗原抗体效价均能达到1∶12 800。本实验为后期免疫阻断效应研究、多时期多价疫苗的构建及鼠疫模型的建立提供了实验基础。更多还原

【Abstract】 This study obtains candidate antigens Pys25(217AA)andPys48(455AA)of plasmodium yoelii in perfect stage through silkworm baculovirus surface display technology so as to establish a new rodent model.It first uses escherichia coli pronucleus expression system to express both antigens and uses immune Balb/c mice to prepare polyclonal antibody,meanwhile uses silkworm baculovirus surface display technology and transforms pFastBac Dual carrier,adds mammal promoter CMV at the front end of its Ph promoter and conducts fusion expression of target protein gene and baculovirus envelope protein gp64 gene segment.Fusion protein is displayed on the surface of virus particle and used to immunize Balb/c mice.In the body of mice,CMV promoter activates fusion protein expression and jointly stimulates mice to produce polyclonal antibody.According to indirect ELISA detection,the valence of antibody of both antigens can reach 1:12800.This experiment provides an experimental basis for later study on immune blocking effect and the establishment of multi-period polyvalent vaccine and plague model.更多还原