摘要
为研究迟缓爱德华菌外膜蛋白OmpA免疫保护性,本研究利用PCR方法扩增迟缓爱德华菌ompA基因,构建重组载体pET-32a-ompA,将其转化大肠杆菌BL21后诱导表达,表达产物经SDS-PAGE和western blot分析显示,重组蛋白大小约58 ku;将纯化的重组蛋白免疫小鼠后,以迟缓爱德华菌强毒株ET-13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55%。本研究克隆了迟缓爱德华菌ompA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OmpA蛋白亚单位疫苗的研制奠定基础。
To study the immunoprotection of OmpA against Edwardsiella tarda in mice, the encoding gene, ompA, was amplified by PCR from E.tarda ET-13 and cloned into the pET-32a vector. The recombinant plasmid was transformed into E.coli BL21 （DE3） cells, in which a recombinant OmpA protein （rOmpA） was expressed by inducing with IPTG. SDS-PAGE and western blot analysis showed that the expressed rOmpA was about 60ku which was recognized by the positive serum against E.tarda. The mice were immunized intramuscularly with purified rOmpA, and challenged with the E.tarda ET-13. Result showed that the rOmpA provided a 55% protection for the immunized mice. The data demonstrated that the OmpA could be used as the antigen for developing the subunit vaccine.
作者
张志强
杨楠
李永慧
王洪彬
冯东青
吴同垒
史秋梅
朱国强
ZHANG Zhi-qiang;YANG Nan;LI Yong-hui;WANG Hong-bin;FENG Dong-qing;WU Tong-lei;SHI Qiu-mei;ZHU Guo-qiang(Hebei Key Laboratory of Preventive Veterinary Medicine, Hebei Normal University of Science ＆Technology, Qinhuangdao 066004, China;The Second Hospital of Qinhuangdao, Qinhuangdao 066604, China;Yangzhou university, Yangzhou 225009, China)
出处
《中国预防兽医学报》
CAS
CSCD
北大核心
2018年第6期534-537,共4页
Chinese Journal of Preventive Veterinary Medicine
基金
中国博士后科学基金(2017M611935)
河北省科技厅奖励性后补助项目(15926620H)
秦皇岛市科技局科技支撑计划(201602A341)
河北科技师范学院博士启动基金(2015YB002)
关键词
迟缓爱德华菌
外膜蛋白
OMPA
蛋白表达
免疫保护力
Edwardsiella tarda
outer membrane protein
OmpA
protein expression
immunological protection
