猪流行性腹泻病毒N基因的克隆及原核表达  

Cloning and expression of porcine epidemic diarrhea virus N gene

作　　者：杨巍[1] 李广兴[1] 孟凡丹[1] 王鑫[1] 任晓峰[1] 

机构地区：[1]东北农业大学动物医学学院,哈尔滨150030

出　　处：《东北农业大学学报》2011年第6期81-85,I0001,共6页Journal of Northeast Agricultural University

基　　金：黑龙江省教育厅"黑龙江省普通高等学校新世纪优秀人才培养计划"项目(1155--NCET--005)

摘　　要：从患猪流行性腹泻的病猪粪便中分离了猪流行性腹泻病毒(PEDV)命名为HLJBY,将病毒在Vero细胞上传代,使之适应生长。将扩增的猪流行性腹泻病毒的N基因克隆到原核表达载体PET30a上,构建了重组表达质粒PET-30a-PEDV-N,并在37℃的条件下诱导表达,通过不同时间取样,经SDS-PAGE分析结果表明,该蛋白获得了高效的表达。将PEDV在Vero细胞上大量繁殖后经超离纯化,作为免疫原制备兔抗PEDV的多克隆抗体血清。Western-blot结果表明蛋白与多抗血清具有良好的免疫反应性。The porcine epidemic diarrhea virus（PEDV） that was isolated from the feces of pigs with diarrhea was named HLJBY and was passaged in Vero cell line.The amplified PEDV N gene was cloned into prokaryotic expression vector PET30a,constructing a recombinant expression vector PET-30a-PEDV-N.High level expression of the N protein was analyzed by SDS-PAGE at different time points at 37 ℃ after IPTG induction.The cultured viruses were purified by ultracentrifugation and injected to a rabbit to generate polyclonal antibody.Western-blotting result showed the polyclonal antibody could specifically bind to PEDV N protein,which indicated that the recombinant fusion protein had excellent immunogenicity.

关 键 词：PEDV N基因 原核表达 N蛋白 

分 类 号：S826[农业科学—畜牧学]