仙台病毒核蛋白的原核表达及鉴定  

PROKARYOTIC EXPRESSION AND DETECTION OF NUCLEOPROTEIN OF SENTAI VIRUS

作　　者：魏晓锋 胡建华 高诚 

机构地区：[1]上海实验动物研究中心上海市实验动物质量监督检验站,上海201203

出　　处：《中国动物传染病学报》2010年第4期32-36,共5页Chinese Journal of Animal Infectious Diseases

摘　　要：本研究从一株鼠仙台病毒(Sendai virus,SeV)中扩增获得核蛋白(nucleocapsid protein,NP)基因,将其克隆入pET32a(+)中,转化大肠杆菌BL21(DE3)中诱导表达。将原核表达产物作为免疫原,免疫BALB/C小鼠,制备获得抗NP的特异性多抗,经间接免疫荧光法和Western blot法检测,该表达产物原具有良好的免疫原性。将该产物纯化作为包被抗原,建立了一种检测SeV感染的间接ELISA方法。通过H1N1流感病毒、H9N2流感病毒、新城疫病毒阳性血清进行交叉试验表明:该间接ELISA方法有很好的特异性。这为实验动物仙台病毒抗体的检测提供了一种快速、简便的血清学诊断方法。In the present study,nucleoprotein（NP） gene of a murine Sendai virus（SeV） was amplified and subcloned into pET32a（＋） vector for prokaryotic expression.The recombinant NP proteins were purified and used to immunize mice.The mouse antiserum samples reacted in immunofluorescence assay（IFA） and Western blot analysis,confirming the immunogenicity of the recombinant proteins.Then,the purified proteins were used to coating ELISA plates for detection of mouse antibody level.The coating antigen did not react with antibodies against H1N1 influenza virus,H9N2 Influenza virus and Newcastle disease virus（NDV）,indicating that the ELISA method can be used to detect antibodies to Sendai virus in mice.

关 键 词：仙台病毒 核蛋白 原核表达 ELISA 

分 类 号：S852.659.5[农业科学—基础兽医学] R446.11[农业科学—兽医学]