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
由昭红;陈冲;
摘要(Abstract):
[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。
关键词(KeyWords): F18菌毛;分离株;免疫原性
基金项目(Foundation):
作者(Author): 由昭红;陈冲;
Email:
DOI: 10.13989/j.cnki.0517-6611.2009.24.136
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