中国现代医学杂志2016，Vol.26Issue(18)：18-21,4.DOI:10.3969/j.issn.1005-8982.2016.18.004
人致病性呼肠孤病毒M3编码蛋白纯化及多克隆抗体制备
Purification of human pathogenic reovirus M3 gene encoded protein and preparation of its polyclonal antibody
摘要
Abstract
Objective To prepare an anti-NBVs (Nelson Bay orthoreoviruses) M3 polyclonal antibody according to the human pathogenic NBVs M3 gene. Methods NBVs M3 plasmid-Pris His MB-M3 was built and transformed into E. coli BL21 (DE3) competent cells. Isopropyl-β-D-thiogalactoside (IPTG) was used to induce protein expression, the NBVs M3 fusion protein was obtained by the method of Ni-NTA agarose; at the same time, the purified fusion protein was utilized as an antigen to immunize rabbits to obtain anti-NBVs M3 polyclonal antibody. The accuracy of the protein and polyclonal antibody resistance were detected by Western blot. Results SDS-PAGE electrophoresis and Coomassie blue staining showed that when the induction was carried out with 0.3 mmol/L IPTG at 25℃for 12 h, the fusion protein expression was the highest. When the imidazole concentration was MCAC-20 and MCAC-40, the fusion protein could be obtained. Western blot showed anti-NBVS M3 polyclonal antibody was successfully prepared with strong specificity when the purified fusion protein was used to immune rabbits. Conclusions A highly sensitive and specific anti-NBVs M3 polyclonal antibody has been successfully obtained. It has a high application value for further study of the pathogenicity of the viruses.关键词
Pris His MB- M3质粒/纳尔逊海湾病毒(NBVs)/纯化蛋白/多克隆抗体Key words
Pris His MB-M3 plasmid/Nelson Bay orthoreoviruses (NBVs)/purified protein/polyclonal antibody分类
医药卫生引用本文复制引用
陈江曼,李永刚,王佳美,佟伟,任雨舒,李新..人致病性呼肠孤病毒M3编码蛋白纯化及多克隆抗体制备[J].中国现代医学杂志,2016,26(18):18-21,4.基金项目
辽宁医学院领军人物人畜共患病创新团队（No173615007）；锦州医科大学校长基金-奥鸿博泽研究生科技创新基金 （）
