| 猪瘟病毒E2蛋白A/D区单抗的制备及其抗原表位的初步研究 |
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| 引用本文： | 张春玲,宗先伟,杨文超,张敬梅,彭明义,程宝艳,余为一.猪瘟病毒E2蛋白A/D区单抗的制备及其抗原表位的初步研究[J].中国畜牧兽医,2009,36(9):50-54. |
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| 作者姓名： | 张春玲 宗先伟 杨文超 张敬梅 彭明义 程宝艳 余为一 |
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| 作者单位： | （1.安徽农业大学动物科技学院， 合肥 230036； 2.山东六和集团有限公司， 德州 253000；3.安徽省农业科学院畜牧兽医研究所， 合肥 230031） |
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| 基金项目： | 安徽省"十五"科技攻关项目 |
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| 摘 要： | 应用分子克隆技术将猪瘟病毒E2蛋白部分基因插入到载体pGEX-4T-1中构建重组质粒pGEX-4T-E（A），在大肠杆菌Rosetta中表达融合蛋白GST-E（A），以纯化的融合蛋白免疫BALB/c小鼠，取其脾细胞与骨髓瘤细胞SP2/0进行融合，经克隆化和间接ELISA筛选，获得了C3、A1两株稳定分泌抗猪瘟病毒E2蛋白单克隆抗体的杂交瘤细胞株，ELISA结果显示，其腹水效价为1∶128000和1∶256000。Western blotting结果证实，单克隆抗体C3和A1能与猪瘟病毒（classical swine fever virus，CSFV）发生特异性的反应，表明该单抗是针对猪瘟病毒E2蛋白的保守线性表位。
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| 关 键 词： | 猪瘟病毒 E2蛋白 单克隆抗体 鉴定 |
Preparation and Identification of Monoclonal Antibodies against the Truncated E2 Protein of Classical Swine Fever Virus |
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| ZHANG Chun-ling,ZONG Xian-wei,YANG Wen-chao,ZHANG Jing-mei,PENG Ming-yi,CHENG Bao-yan,YU Wei-yi.Preparation and Identification of Monoclonal Antibodies against the Truncated E2 Protein of Classical Swine Fever Virus[J].China Animal Husbandry & Veterinary Medicine,2009,36(9):50-54. |
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| Authors: | ZHANG Chun-ling ZONG Xian-wei YANG Wen-chao ZHANG Jing-mei PENG Ming-yi CHENG Bao-yan YU Wei-yi |
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| Institution: | （1.College of Animal Science and Technology, Anhui Agricultural University, Hefei 230036, China; 2.Shandong Liuhe Poultry Company， Dezhou 253000, China; 3.Institute of Animal and Veterinary Science, Anhui Academy of Agricultural Science, Hefei 230031, China） |
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| Abstract: | A truncated gene encoding the major antigenic domains of E2 protein of classical swine fever virus(CSFV) was amplified and cloned into pGEX-4T-1 expression vector to obtain recombinant pGEX-4T-E(A).The recombinant GST-E(A)protein expressed in E.coli Rosetta.BALB/c mice was immunized with purified GST-E(A)protein as antigen and mouse splenic cells were fused with SP2/0 cells.Hybridoma cells were screened by ELISA and subcloned.Two hybridoma cells C3 and A1 secreting anti-CSFV monoclonal antibodies were obtai... |
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| Keywords: | classical swine fever virus E2 protein monoclonal antibody identification |
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