摘要
为研究迟缓爱德华菌鞭毛蛋白FlgJ的免疫保护性,本研究利用PCR方法扩增迟缓爱德华菌flgJ基因,构建重组载体pET-32a-flgJ,将其转化大肠杆菌BL21后进行诱导表达,表达产物经SDS-PAGE和Western blot分析显示,重组蛋白大小约54 000;将纯化的重组蛋白免疫小鼠后,以迟缓爱德华菌分离株ET-13进行攻毒,结果显示该重组蛋白对免疫组小鼠具有保护力,保护率为70%。本研究克隆了迟缓爱德华菌外膜蛋白flgJ基因并表达了相应重组蛋白,免疫小鼠后能够提供一定保护,为重组FlgJ蛋白亚单位疫苗的研制奠定基础。
To study the immunoprotection of FlgJ against Edwardsiella tardain mice,the encoding gene,flgJ,was amplified by PCR fromE.tardaisolate ET-13,and furthered cloned into the pET-32 avector.The recombinant plasmid was transformed into E.coli BL21(DE3)cells,in which a recombinant FlgJ(rFlgJ)protein was expressed by inducing with IPTG.SDS-PAGE and Western blot analysis showed that the expressed rFlgJ was about 54 000,which was recognized by the positive serum against E.tarda.The mice were immunized intramuscularly with purified rFlgJ,and challenged with the E.tardaisolate ET-13.Result showed that the rFlgJ provided a 70% protection for immunized mice.The data demonstrated that the FlgJ could be used as the antigen for developing the subunit vaccine
作者
张志强
杨楠
李永慧
苏硕青
冯东青
吴同垒
高桂生
钱爱东
史秋梅
ZHANG Zhi-qiang;YANG Nan;LI Yong-hui;SU Shuo-qing;FENG Dong-qing;WU Tong-lei;GAO Gui-sheng;QIAN Ai-dong;SHI Qiu-mei(Hebei Normal University of Science Technology,Qinhuangdao,Hebei 066000,China;The Second Hospital of Qinhuangdao ,Qin-huangdao,Hebei066600,China;College of Animal Science and Technology,J ilin Agricultural University,Changchun 130000,China)
出处
《中国兽医学报》
CAS
CSCD
北大核心
2018年第12期2348-2351,2359,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基金资助项目(15926620H)
秦皇岛市科技局科技支撑计划资助项目(201602A341)
河北科技师范学院博士启动基金资助项目(2015YB002)
关键词
迟缓爱德华菌
外膜蛋白
FlgJ
蛋白表达
免疫保护力
Edwardsiella tarda
outer membrane protein
FlgJ
protein expression
immunological protection
