牛病毒性腹泻病毒E2蛋白的多克隆抗体制备及鉴定  

Preparation and characterization of antisera against recombinant E2 protein of bovine viral diarrhea virus

作　　者：高欲燃[1,2] 朱远茂[1] 康健[3] 史鸿飞[1] 李娇[1] 任宪刚[1] 冯军科[1] 于作[1] 薛飞[1] 

机构地区：[1]中国农业科学院哈尔滨兽医研究所兽医生物技术国家重点实验室/大动物传染病研究室,黑龙江哈尔滨150001 [2]东北农业大学动物医学学院,黑龙江哈尔滨150030 [3]南京农业大学动物医学院,江苏南京210095

出　　处：《中国预防兽医学报》2011年第3期227-231,共5页Chinese Journal of Preventive Veterinary Medicine

基　　金：黑龙江省十一五科技攻关计划项目(GA06B202-3);兽医生物技术国家重点实验室基本科研业务费(NKLVBP200816)

摘　　要：为制备牛病毒性腹泻病毒(BVDV)重组E2蛋白的兔源多克隆抗体,本研究利用表达BVDV E2蛋白的重组质粒pET30a-E2转化E.coli BL21(DE3),经诱导表达获得重组E2蛋白。Western blot检测显示纯化蛋白能够与BVDV参考阳性血清反应。以纯化的重组E2蛋白免疫新西兰白兔制备多克隆抗体,病毒中和试验测定其中和效价为1:2048,间接免疫荧光和western blot试验表明其具有良好的反应性和特异性。本研究制备的BVDV重组E2蛋白兔源多克隆抗体可应用于BVDV的检测,同时为进一步建立检测BVDV E2蛋白的ELISA方法奠定基础。To prepare the polyclonal antibody against recombinant E2 protein of bovine viral diarrhea virus（BVDV） in rabbits,E.coli BL21（DE3） was transformed with the recombinant plasmid pET30a-E2.The recombinant E2 protein was expressed in E.coli after cultivation and induction.The purified recombinant E2 protein could be recognized by specific BVDV antisera in western blot.Then the purified recombinant E2 protein was used as antigen for immunizing rabbit to prepare polyclonal antibody against the recombinant E2 protein.The result of virus neutralization test showed that the titer of the polyclonal antibody to neutralize BVDV was 1：2,048.The polyclonal antibody against the recombinant E2 protein of BVDV also had highly reactivity and specialty in immunofluorescence analysis and western blot.The polyclonal antibody against recombinant E2 protein of BVDV developed in rabbits could be used in detection of BVDV in China and provided a good basis for establishing an ELISA for detecting of E2 protein of BVDV.

关 键 词：牛病毒性腹泻病毒 E2蛋白 多克隆抗体 

分 类 号：S852.5[农业科学—基础兽医学]