猪流行性腹泻病毒中和抗原表位基因的原核表达、纯化及免疫原性鉴定  

Prokaryotic expression,purification and identification of immunogenicity of the core neutralizing epitope gene of Porcine epidemic diarrhea virus

作　　者：满坤[1,2] 杨兵[2] 覃湘婕[2] 周宏专[2] 张晓东[2] 徐福洲[2] 王金洛[2] 

机构地区：[1]北京农学院,北京102206 [2]北京市农林科学院,北京100097

出　　处：《黑龙江畜牧兽医》2013年第11期26-29,共4页Heilongjiang Animal Science And veterinary Medicine

基　　金：北京市农林科学院一般项目(2010A000)

摘　　要：为了获得具有免疫原性的猪流行性腹泻病毒(PEDV)主要中和抗原表位(COE)蛋白及其多克隆抗体,试验对已发表的PEDV基因序列进行比对,并设计、合成COE编码基因(420 bp),将其克隆入原核表达载体pET-30a,转化到大肠杆菌BL21(DE3)感受态细胞中,IPTG诱导表达,利用胶体金和Western-blot方法检测表达产物;纯化后免疫兔制备高免血清,并利用ELISA方法检测血清抗体效价。结果表明:表达的重组蛋白可与PEDV特异性抗体结合;免疫兔后获得的血清抗体效价达1∶6 400,具有免疫原性。To obtain the immunogenic core neutralizing epitope （COE） protein of Porcine epidemic diarrhea virus （PEDV） that has immunoge- nicity and its polyclonal antibody. According to the published PEDV gene sequences, the COE encoding gene （420 bp） was designed and syn- thesized, and then was cloned into the prokaryotic expression vector pET - 30a. The recombinant plasmid was transformed into E. coil BI21 （ DE3 ） to induce the protein expression by IPTG, and then the expressed protein was detected by Western - blot and immune colloidal gold as- say. After purified,the protein was used to immunize rabbit to prepare anti - serum, and then the titer of the anti - serum was detected by ELISA. The ~sults showed that the expressed recombinant protein could combine with the specific antibody of PEDV ,and the titer of the anti - serum was 1 ： 6 400. The results indicate that the COE protein of PEDV has immunogenicity.

关 键 词：猪流行性腹泻病毒 中和抗原表位 原核表达 纯化 免疫原性 

分 类 号：S852.43[农业科学—基础兽医学]