| 猪瘟病毒E0蛋白的原核表达及多克隆抗体的制备 |
| |
| 引用本文： | 贾俊杰,张必凯,张莉,谢金鑫,郭焕成,龚文杰,涂长春.猪瘟病毒E0蛋白的原核表达及多克隆抗体的制备[J].动物医学进展,2016(5). |
| |
| 作者姓名： | 贾俊杰 张必凯 张莉 谢金鑫 郭焕成 龚文杰 涂长春 |
| |
| 作者单位： | 军事医学科学院 军事兽医研究所，吉林省人兽共患病预防与控制重点实验室，吉林长春 130122 |
| |
| 基金项目： | 中国博士后科学基金项目(2013M532129)；国家自然科学基金项目(31130052；31572528) |
| |
| 摘 要： | 为表达猪瘟病毒E0蛋白并制备其多克隆抗体,本研究构建E0蛋白的原核表达载体,转化至BL21(DE3)菌株,IPTG诱导表达,亲和层析及切胶回收纯化重组蛋白。SDS-PAGE和Western blot分析显示E0重组蛋白主要以包涵体形式表达,亲和层析纯化获得了E0重组蛋白,用其免疫Balb/c小鼠4次制备E0重组蛋白的多克隆抗体。间接ELISA显示,免疫小鼠血清中E0蛋白抗体效价为1∶50 000。获得的E0蛋白多抗能与病毒感染细胞及E0-EGFP融合表达细胞中天然结构的E0蛋白发生特异性反应。本研究制备的E0重组蛋白及其多克隆抗体为进一步研究E0蛋白的功能和免疫原性奠定了基础。
|
| 关 键 词： | 猪瘟病毒 E0蛋白 原核表达 多克隆抗体 |
Prokaryotic Expression of E0 Protein of Classical Swine Fever Virus and Preparation of Its Polyclonal Antibodies |
| |
| Abstract: | To express E0 protein of CSFV and prepare its polyclonal antibodies (pAb),the recombinant E0 protein of CSFV Shimen strain was expressed in E.coli BL21(DE3)with IPTG induction and successively purified by affinity chromatography and gel slice.Balb/c mice were immunized four times with the purified recombinant E0 protein.SDS-PAGE and Western blot analysis showed that recombinant protein was ex-pressed mainly in the inclusion body.The indirect ELISA showed that the antibody titers in the serum of immunized mice was at 1:50000.Moreover,the obtained anti-E0 pAb can specifically react with the E0 pro-tein expressed in virus-infected cells and E0-EGFP-expressing cells.The obtained E0 protein and its poly-clonal antibodies in this study will be useful for future studies on the function and immunogenicity of CSFV E0 protein. |
| |
| Keywords: | Classical swine fever virus prokaryotic expression E0 protein polyclonal antibody |
| 本文献已被 CNKI 万方数据 等数据库收录！ |
|
