| 鲤细胞因子多克隆抗体的制备及检测 |
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| 引用本文： | 冯军厂,常绪路,朱振祥,郭祥瑞,赵燕静,刘慧芬,张建新,聂国兴.鲤细胞因子多克隆抗体的制备及检测[J].水产学报,2018,42(10):1615-1625. |
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| 作者姓名： | 冯军厂 常绪路 朱振祥 郭祥瑞 赵燕静 刘慧芬 张建新 聂国兴 |
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| 作者单位： | 河南师范大学水产学院河南省水产动物养殖工程技术研究中心水产动物疾病控制河南省工程实验室 |
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| 基金项目： | 河南省创新型科技团队支持计划（CXTD2016043）；河南师范大学博士启动基金（qd16155）；河南省重点科技攻关项目（172102110195，172102210343）；河南省高等学校重点科研项目（17B240002，16A240004）；河南师范大学校青年基金（2016QK23） |
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| 摘 要： | 为从蛋白质水平研究细胞因子在鲤体内免疫应答过程中的合成变化,本研究采用PCR技术克隆TNF-α、IL-1β、IL-6、IL-12、IL-10和TGF-β基因含有部分抗原决定簇的片段,引入双酶切位点Bam HⅠ和HindⅢ后连接至p ET-32a/21a,构建相应的表达载体,制备多克隆抗体。采用ELISA检测抗体效价,并以此作为实验工具,检测经嗜水气单胞菌感染后鲤血清中炎性细胞因子的合成变化。结果显示,基因TNF-α、IL-1β、IL-6、IL-12、IL-10和TGF-β融合蛋白分子量分别约为31.8、31.7、35.3、32.5、18.0和33.6 ku;抗体效价达到2.4×106;在病原菌感染后的不同阶段,促炎细胞因子TNF-α、IL-1β、IL-6、IL-12和抗炎细胞因子IL-10、TGF-β呈现出不同的合成变化。研究表明,制备的抗体具有较高的效价、亲和力和特异性,可用于鲤细胞因子的定量研究,该抗体的获得为鲤免疫应答与细胞因子合成的系统研究奠定了基础。同时,获取的鲤细胞因子TNF-α、IL-1β、IL-6、IL-12、IL-10和TGF-β的抗体亦可用于其他鱼类细胞因子蛋白质水平的定量研究。
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| 关 键 词： | 鲤 细胞因子 原核表达 抗体效价 病原菌 ELISA |
| 收稿时间： | 2017/10/13 0:00:00 |
| 修稿时间： | 2017/12/25 0:00:00 |
Preparation and detection of cytokines polyclonal antibodies of Cyprinus carpio |
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| FENG Junchang,CHANG Xulu,ZHU Zhenxiang,GUO Xiangrui,ZHAO Yanjing,LIU Huifen,ZHANG Jianxin and NIE Guoxing.Preparation and detection of cytokines polyclonal antibodies of Cyprinus carpio[J].Journal of Fisheries of China,2018,42(10):1615-1625. |
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| Authors: | FENG Junchang CHANG Xulu ZHU Zhenxiang GUO Xiangrui ZHAO Yanjing LIU Huifen ZHANG Jianxin and NIE Guoxing |
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| Institution: | Engineering Lab of Henan Province for Aquatic Animal Disease Control, Engineering Technology Research Center of Henan Province for Aquatic Animal Cultivation, College of Fisheries, Henan Normal University, Xinxiang 453007, China,Engineering Lab of Henan Province for Aquatic Animal Disease Control, Engineering Technology Research Center of Henan Province for Aquatic Animal Cultivation, College of Fisheries, Henan Normal University, Xinxiang 453007, China,Engineering Lab of Henan Province for Aquatic Animal Disease Control, Engineering Technology Research Center of Henan Province for Aquatic Animal Cultivation, College of Fisheries, Henan Normal University, Xinxiang 453007, China,Engineering Lab of Henan Province for Aquatic Animal Disease Control, Engineering Technology Research Center of Henan Province for Aquatic Animal Cultivation, College of Fisheries, Henan Normal University, Xinxiang 453007, China,Engineering Lab of Henan Province for Aquatic Animal Disease Control, Engineering Technology Research Center of Henan Province for Aquatic Animal Cultivation, College of Fisheries, Henan Normal University, Xinxiang 453007, China,Engineering Lab of Henan Province for Aquatic Animal Disease Control, Engineering Technology Research Center of Henan Province for Aquatic Animal Cultivation, College of Fisheries, Henan Normal University, Xinxiang 453007, China,Engineering Lab of Henan Province for Aquatic Animal Disease Control, Engineering Technology Research Center of Henan Province for Aquatic Animal Cultivation, College of Fisheries, Henan Normal University, Xinxiang 453007, China and Engineering Lab of Henan Province for Aquatic Animal Disease Control, Engineering Technology Research Center of Henan Province for Aquatic Animal Cultivation, College of Fisheries, Henan Normal University, Xinxiang 453007, China |
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| Abstract: | |
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| Keywords: | Cyprinus carpio cytokines prokaryotic expression antibody titer pathogenic bacteria ELISA |
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