迟缓爱德华菌菌毛蛋白原核表达及多抗血清制备  

Prokaryotic expression of PILI protein of Edwardsiella tarda and preparation of polyclonal antibody

作　　者：张志强[1] 杨楠[1] 李永慧 潘晓艺[3] 单晓枫[4] 康元环[4] 焦贺静 韩红升 吴同垒[1] 高桂生[1] 

机构地区：[1]河北科技师范学院河北省预防兽医学重点实验室,河北秦皇岛066004 [2]秦皇岛第二医院,河北昌黎066600 [3]浙江省淡水水产研究所,浙江湖州214000 [4]吉林农业大学动物科学技术学院,吉林长春130000 [5]秦皇岛粮丰海洋生态科技开发股份有限公司,河北秦皇岛066004

出　　处：《中国兽医学报》2017年第9期1705-1709,共5页Chinese Journal of Veterinary Science

基　　金：中国博士后科学基金资助项目(2017M611935);河北省科技厅奖励性后补助资助项目(15926620H);秦皇岛市科技局科技支撑计划资助项目(201401A067;201402B043);河北科技师范学院博士启动基金资助项目(2015YB002);浙江省重大科技专项重点农业资助项目(2012C12009-4)

摘　　要：为研究迟缓爱德华菌菌毛蛋白PILI的功能和免疫原性,本研究利用PCR方法从迟缓爱德华菌基因组中克隆出pili基因,并将其构建到pET-32a原核表达载体上,重组载体转化BL21大肠杆菌诱导重组蛋白表达,通过SDSPAGE和Western blot方法验证重组蛋白表达。用亲和层析方法纯化蛋白,纯化的蛋白免疫小鼠,所得的多抗血清能够特异性识别迟缓爱德华菌PILI蛋白,为研究PILI蛋白奠定基础。To investigate the roles of PILl protein in bacterial adhesion host cells and immunogenic- ity. pill gene was cloned from Edwardsiella tarda strain isolated from farmed turbots,double di- gested,and ligated with expression vector pET-32a. The recombination plasmid pET-32a-pili was transformed into E. coli BL21 strain for protein expression, and identified by SDS-PAGE and Western blot analysis. The collected cells were lyzed by ultraphonic, purified with Ni agrose and thereby used for mouse vaccine. The obtained polyclonal serum specifically bound to bacterial PILI protein in E. tarda as demonstrated by Western blot assay,and may help to analyze the biological roles of PILL in E. tarda.

关 键 词：迟缓爱德华菌 菌毛蛋白 PILI 蛋白表达 纯化 

分 类 号：S852.61[农业科学—基础兽医学] R535[农业科学—兽医学]