摘要
Bfr(Bacterioferritin)是细菌主要的储铁蛋白和重要的免疫相关蛋白。为探索哈维氏弧菌Vibrio harveyi Bfr蛋白的免疫特性,本研究利用大肠杆菌Escherichia coli表达系统表达哈维氏弧菌Bfr重组蛋白(rBfr),并进行纯化,进一步通过动物模型评估rBfr的免疫特性。结果显示,成功表达了哈维氏弧菌Bfr蛋白,Bfr蛋白分子量大小约为25 ku;表达的重组蛋白主要以可溶性形式存在;免疫小鼠结果显示,rBfr蛋白能激发小鼠产生高水平的特异性抗体,rBfr具有较好的免疫原性;利用斑马鱼Danio rerio建立哈维氏弧菌感染模型,表明哈维氏弧菌对斑马鱼无致病性。本研究证实了哈维氏弧菌Bfr具有免疫原性,为进一步研制哈维氏弧菌亚单位疫苗和核酸疫苗提供了参考和借鉴。
The recombinant protein rBfr(bacterioferritin)as an immune-related protein and major iron storage protein was expressed in Vibrio harveyi strain VHCL-1 by using Escherichia coli expression system,and rBfr immunological characteristics were evaluated by animal model to investigate the immunological characteristics of Bfr.The recombinant protein rBfr was successfully expressed as soluble form in E.coli BL21 and purified with the molecular weight of 25 ku.The purified rBfr as an immunogen had excellent immunogenicity with a high-level specific antibody determined in the vaccine mice.In addition,a zebrafish(Danio rerio)model was established by Vibrio harveyi infection test,indicating that the expressed Vibrio harveyi strain VHCL-1 is not pathogenic to zebrafish,which provide guidance and foundation for development of subunit vaccine and nucleic acid vaccine of Vibrio harveyi.
作者
张志强
王苗
杜万年
刘勃兴
康元环
单晓枫
吴同垒
朱国强
史秋梅
ZHANG Zhiqiang;WANG Miao;DU Wannian;LIU Boxing;KANG Yuanhuan;SHAN Xiaofeng;WU Tonglei;ZHU Guoqiang;SHI Qiumei(Hebei Key Laboratory of Preventive Veterinary Medicine,Hebei Normal University of Science&Technology,Qinhuangdao 066000,China;College of Animal Science and Technology,Jilin Agriculture University,Changchun 130000,China;Yangzhou University,Yangzhou 225009,China)
出处
《水产学杂志》
CAS
北大核心
2021年第2期23-27,共5页
Chinese Journal of Fisheries
基金
农业部淡水渔业健康养殖重点实验室、浙江省鱼类健康与营养重点实验室或浙江省淡水水产遗传育种重点实验室开放基金课题(ZJK201812)
中国博士后科学基金(2017M611935)
河北科技师范学院海洋专项(2018HY007).
关键词
哈维氏弧菌
BFR
免疫特性
Vibrio harveyi
bacterioferritin(Bfr)
immunological characteristics
