鼠伤寒沙门氏菌鞭毛蛋白提取及多抗制备  

Extraction of flagellin from Salmonella typhimurium and preparation of its polyclonal antibody

作　　者：颜成英[1,2] 汤晓艳[1] 王敏[1] 康大成 李朋颖 龚艳[1] 

机构地区：[1]中国农业科学院农业质量标准与检测技术研究所,农业部农产品质量安全重点实验室,北京100081 [2]南京农业大学,教育部肉品加工与质量控制重点实验室,江苏南京210095

出　　处：《食品工业科技》2014年第22期176-178,183,共4页Science and Technology of Food Industry

基　　金："十二五"国家科技支撑计划项目(2012BAD28B03);国家自然青年科学基金项目(31000799);国家现代农业产业技术体系专项(CARS-42)

摘　　要：鼠伤寒沙门氏菌是沙门氏菌属常见致病菌,其鞭毛蛋白具有抗原性,可诱导免疫系统产生针对该蛋白的抗体。实验采用直接提取法获得鼠伤寒沙门氏菌鞭毛蛋白,并以该蛋白为抗原免疫小鼠,制备鞭毛蛋白多抗,使用间接ELISA方法测定多抗的效价和特异性。SDS-PAGE实验结果表明,蛋白条带单一,分子量大小为57ku,说明提取的鞭毛蛋白是目标蛋白且纯度较高;间接ELISA检测结果表明,制备多抗的效价为1∶102400,除与猪霍乱沙门氏菌发生交叉反应外,与其他6种沙门氏菌和6种常见非沙门氏菌属的致病菌均不发生反应,说明该抗体的效价较高,特异性较好,可以用于与磁珠的偶联,为免疫磁珠的制备打下基础。Salmonella typhimurium is the most common pathogenic bacterium in salmonella and had flagellin of antigenicity which could induce immune system to produce antibody. Our research adopted direct extract method to get flagellin of salmonella,and used this protein as antigen to immune mice for flagellin polyclonal antibody. In this research,valence and specificity of antibody were tested through indirect ELISA testing. SDSPAGE experiment showed protein stripe was single and molecular weight was 57 ku,which means extracted flagellin was the target protein and it＇s of high purity. ELISA testing indicated that the valence of antibody was1 ∶ 102400 and it had cross reaction with Salmonella choleraesuis only while it didn ＇ t react with other 6salmonella and 6 common pathogenic bacteria of non-salmonella. It stated this antibody had high valence and good specificity and could be used to couple with magnetic bead.

关 键 词：鼠伤寒沙门氏菌 鞭毛蛋白 多抗 

分 类 号：TS251.7[轻工技术与工程—农产品加工及贮藏工程]