日本血吸虫PROHIBITIN 2克隆和表达及免疫保护效果评估  

Cloning and Expression of Schistosoma japonicum Prohibitin 2 and Evaluation of Immunoprotective Effect

作　　者：杨珊珊 杨静云 张旻[2] 张倩[1] 何静 黄明月 窦记晨 洪炀[1] 林矫矫[1] 傅志强[1] YANG Shanshan;YANG Jingyun;ZHANG Min;ZHANG Qian;HE Jing;HUNG Mingyue;DOU Jichen;HONG Yang;LIN Jiaojiao;FU Zhiqiang(Key Laboratory of Animal Parasitology,Ministry of Agriculture and Rural Affairs,Shanghai Veterinary Research Institute,CAAS,Shanghai 200241,China;College of Animal Science and Technology,Henan University of Science and Technology,Luoyang 471000,China)

机构地区：[1]中国农业科学院上海兽医研究所农业农村部动物寄生虫学重点实验室,上海200241 [2]河南科技大学动物科技学院,洛阳471000

出　　处：《中国动物传染病学报》2022年第6期83-90,共8页Chinese Journal of Animal Infectious Diseases

基　　金：国家自然基金(31872256);国家重点研发计划2017专项(2017YFD0501306);河南科技大学博士科研启动基金(13480069)。

摘　　要：为研究日本血吸虫PROHIBITIN的生物特性,利用PCR和3’RACE扩增到日本血吸虫抗增殖蛋白2(Schistosoma japonicum Prohibitin 2,Sj PHB2)基因,构建重组表达质粒pGEX-4T-1-SjPHB2,并在大肠杆菌(E.coli)中诱导表达,用Western blot检测其免疫原性。将重组蛋白免疫小鼠评估其免疫保护效果,ELISA检测其特异性抗体水平。结果显示:日本血吸虫Sj PHB2基因编码299个氨基酸,含PHB结构域;含GST标签的r Sj PHB2融合蛋白的分子量为59 kDa;r Sj PHB2能被GST单克隆抗体抗和免疫血清识别,表明其具有较好的免疫原性;将r Sj PHB2结合ISA206佐剂免疫BALB/c小鼠可诱导较高水平的特异性IgG抗体,但未能诱导显著的减虫率和减卵率。本文结果为开展日本血吸虫Sj PHB2的表达特性和生物学功能提供了基础。In order to investigate the biological characteristics of Schistosoma japonicum prohibitin 2(PHB2),the gene SjPHB2 was amplifi ed by PCR and 3’RACE and the recombinant plasmid pGEX-4T-1-SjPHB2 was constructed and expressed in E.coli.The BALB/c mice were immunized with the fusion protein r SjPHB2 to evaluate its immunoprotective effect against S chistosomi asis.The immunogenicity of the obtained r SjPHB2 was analyzed in Western blotting and the level of specifi c antibodies was tested in ELISA.The results showed that the SjPHB2 gene encodes 299 amino acids and contained the PHB2 domain.The molecular weight of r SjPHB2 fused with GST tag was 59kDa.The r SjPHB2 was probed by GST-specifi c monoclonal antibodies and immune serum in Western blotting,indicating that it had good immunogenicity.Immunization of BALB/c mice with r SjPHB2 combined with ISA206 adjuvant produced high level of specifi c IgG antibodies but failed to induce signifi cant worm and egg reduction.The results of this study provided a basis for further research of biological functions of SjPHB2.

关 键 词：日本血吸虫 抗增殖蛋白2 克隆 表达 

分 类 号：S852.735[农业科学—基础兽医学]