F18^+ ETEC分离株的鉴定·菌毛蛋白提取及生物活性验证  

Identification of F18^+ ETEC Isolates,Extraction of Their Fimbriae Proteins and Preliminary Verification of Their Biological Activity

作　　者：由昭红[1] 陈冲 

机构地区：[1]枣庄学院,山东枣庄277160 [2]山东省枣庄市兴牧生物科技有限公司,山东枣庄277160

出　　处：《安徽农业科学》2009年第24期11441-11443,共3页Journal of Anhui Agricultural Sciences

摘　　要：[目的]探讨F18+ETEC分离株的鉴定、菌毛蛋白的提取及菌毛蛋白生物活性的初步验证。[方法]分别以分离株A20、D20为模板进行PCR扩增,用1%琼脂糖凝胶电泳检测PCR产物。采用热抽提法分别提取F18标准株和分离株A20、D20的菌毛蛋白,并分别制备弗氏佐剂苗,免疫产蛋鸡,采用间接ELISA法定期检测母鸡的IgY和血清抗体效价。[结果]采用TSB培养基,37℃摇床培养24 h,分离株和标准株菌毛均获得了良好的表达。分离株A20、D20经PCR扩增、琼脂糖凝胶电泳、SDS-PAGE方法鉴定均为F18+ETEC。F18ab分离株和标准株的菌毛对产蛋鸡均具有良好的免疫原性,卵黄中IgY效价最高可达1∶25 600,血清中抗体效价高于卵黄中IgY效价,最高可达1∶51 200。[结论]分离株A20、D20均为F18+ETEC,且对产蛋鸡均具有良好的免疫原性。[Objective] The purpose was to discuss the identification of F18^＋ ETEC isolates,extraction of their imbriae proteins from these strains and preliminary identification of their biological activities. [ Method] With isolates A20 and D20 as the templates, the PCR amplification was carried out and the PCR products were detected by 1% agarose gel electrophoresis. The fimbriae proteins were extracted from standard strains F18 and isolates A20 and D20 by thermal extraction method to prepare Freund＇s adjuvant vaccine and immunize laying hens,resp. The antibody titers of IgY and serum in hens were detected periodically with the indirect ELISA method. [ Result ] The fimbria of isolates and standard strains had a good expression by shaking culture at 37 22 for 24 h on TSB medium. The isolates A20 and D20 were identified as F18^＋ ETEC by PCR amplification, agarose gel electrophoresis as well as SDS-PAGE. All the fimbriaes of isolated and standard strains of F18ab had good immunogenicities on laying hens. The highest IgY titer of yolk was up to 1：25 600,and the antibody titer in serum was higher than that in yolk,which was up to 1：51 200. [ Conclusion] Both of the isolates A20 and D20 were identified as F18^＋ ETEC and had good immunogenicities on laying hens.

关 键 词：F18菌毛 分离株 免疫原性 

分 类 号：S188[农业科学—农业基础科学]