哈维氏弧菌Bfr蛋白的原核表达及免疫特性研究

Prokaryotic Expression and Immunological Characteristics of Bacterioferritin Protein of Vibrio harveyi

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.