微小隐孢子虫cgd8-2500的原核表达及单克隆抗体的制备  

Prokaryotic Expression of Cryptosporidium parvum cgd8-2500 and Preparation of Its Monoclonal Antibody

作　　者：刘雪芹 于志海 孙秋艳 李秀 何孟莲 程光民 王光锋 张风荣 LIU Xue-qin;YU Zhi-hai;SUN Qiu-yan;LI Xiu;HE Meng-lian;CHENG Guang-min;WANG Guang-feng;ZHANG Feng-rong(Shandong Vocational College of Animal Husbandry and Veterinary Medicine,Weifang,Shandong,261061,China)

机构地区：[1]山东畜牧兽医职业学院,山东潍坊261061

出　　处：《动物医学进展》2025年第3期87-92,共6页Progress In Veterinary Medicine

基　　金：中央引导地方科技发展基金项目(YDZX2022137);农牧交错区岗巴羊半舍饲养殖技术的研究与集成示范项目(QYXTZX-RKZ2020-01);山东现代农业家禽产业技术体系疫病防控岗(SDAIT-11-05);无角霍尔巴羊选育与生产性能的测定项目(QYXTZX-RKZ2022-05)。

摘　　要：为研究微小隐孢子虫cgd8-2500蛋白质相关生物学特征,通过生物信息学分析,从微小隐孢子虫基因组中筛选I型跨膜蛋白并选取功能未知、大小适中的基因cgd8-2500;对该基因的氨基酸序列预测分析,选取免疫原性且亲水性好的区域进行原核表达重组蛋白质;利用重组蛋白免疫Balb/c小鼠,制备针对该蛋白的单克隆抗体。结果表明,PCR成功扩增423 bp目的片段,连接T载体,酶切后连至表达载体pET-28a,并获得高纯度蛋白质,免疫Balb/c小鼠后,获得针对cgd8-2500的单克隆抗体。该研究获得了免疫用重组蛋白,进而获得针对cgd8-2500蛋白单克隆抗体,为后期研究其相关生物学特征及其功能提供了工具。To study the biological characteristics of the protein cgd8-2500 related to Cryptosporidium parvum,I-type transmembrane proteins were screened from the Cryptosporidium parvum genome,and the gene cgd8-2500 with unknown function and moderate size was selected.The amino acid sequence of the gene was analyzed by prediction,and the immunogenic and hydrophilic regions were selected for prokaryotic expression recombinant protein.The recombinant protein was used to immunize Balb/c mice,a monoclonal antibody against the protein was prepared.The result showed that,a 423 bp target fragment was successfully amplified by PCR,connected to T vector,and then connected to the expression vector pET-28a after enzyme digestion.High-purity protein was successfully obtained.After immunizing Balb/c mice,a monoclonal antibody against cgd8-2500 was obtained.The successful acquisition of recombinant protein cgd8-2500 and its monoclonal antibody(MAb)provided a reliable tool for elucidating its related biological characteristics and functions in the future.

关 键 词：微小隐孢子虫 重组蛋白cgd8-2500 单克隆抗体 

分 类 号：S855.9[农业科学—临床兽医学]