猪流行性腹泻病毒重组核蛋白作为检测抗原的初步应用  

Preliminary application of the recombinant nucleocapsid protein of porcine epidemic diarrhea virus LJB/03 as detecting antigen in diagnostic assays

作　　者：姜艳平[1] 葛俊伟[1] 汪淼[1] 刘兆磊[1] 裴敦国[1] 韩梓峰[1] 李一经[1] 

机构地区：[1]东北农业大学动物医学院,黑龙江哈尔滨150030

出　　处：《中国预防兽医学报》2008年第8期587-591,共5页Chinese Journal of Preventive Veterinary Medicine

基　　金：黑龙江十一五科技攻关项目(GA06B202-4)

摘　　要：为确定猪流行性腹泻病毒(PEDV)重组N蛋白作为检测抗原的可应用性,将N基因克隆至pProHTa载体,构建重组表达载体,而后转化大肠杆菌BL21感受态细胞并诱导表达。将表达的可溶性融合蛋白经Ni+亲和层析柱纯化,并对其进行Western blot、dot-ELISA、间接ELISA等免疫学实验分析。结果表明,重组N蛋白具有良好的抗原性和较高的敏感性,与其他腹泻病病毒的抗血清无交叉反应。因此,猪流行性腹泻病毒重组N蛋白作为检测抗原在监测病毒感染及评价疫苗免疫效果方面具有较高的应用价值,是代替全病毒作为检测抗原的良好选择。The N gene of porcine epidemic diarrhea virus （PEDV） was cloned into the plasmid pProHTa to construct the recombinant expression vector, Recombinant N protein was expressed in E.coli BL21 by IPTG induction. The soluble His-tagfused N protein was purified through Ni^＋ chromatography column and its antigenicity was validated by Western blot, dot-ELISA and indirect ELISA. The results showed that recombinant protein retained strong antigenicity of the native N protein and was highly specific. It is a valuable antigen alternative for diagnosis of virus infection and evaluation to vaccine efficiency.

关 键 词：猪流行性腹泻病毒 N蛋白 诊断抗原 

分 类 号：S852.659[农业科学—基础兽医学]