| PRRSV Nsp2高变区基因的表达及免疫学特性鉴定 |
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| 引用本文： | 马利宏,肖一红,吕军华,王伟伟,董施伟,周恩民.PRRSV Nsp2高变区基因的表达及免疫学特性鉴定[J].西南农业学报,2011,24(1). |
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| 作者姓名： | 马利宏 肖一红 吕军华 王伟伟 董施伟 周恩民 |
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| 作者单位： | 山东农业大学动物科技学院,山东,泰安,271018 |
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| 基金项目： | 国家自然科学基金，山东省自主创新成果转化重大专项，山东农业大学青年创新基金，山东省博士后创新项目 |
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| 摘 要： | 本研究分别扩增从"高热病"猪体内分离株(TA-12)及美洲经典株ATTC-VR2332 PRRSV Nsp2基因的高变区,亚克隆至表达载体pET-28a中,转化至E.coli BL21中,在IPTG的诱导下进行表达.用SDS-PAGE、Western-blot对表达产物进行鉴定.表达产物纯化后免疫BALB/c小鼠,对其免疫原性进行研究.建立了以纯化的重组蛋白作为包被抗原的间接ELISA,确定了临界值,用建立的ELISA方法并对400份猪血清进行了检测和应用.结果表明,成功的表达了分离株TA-12及ATTC-VR2332 Nsp2基因的高变区.表达蛋白纯化分别免疫小鼠后,获得多克隆抗体,其抗体效价均为1∶100000.优化的ELISA条件为:包被缓冲液为磷酸盐缓冲液,800 ng/孔,血清样品稀释浓度为1∶100.临界值为0.4.血清检测结果表明以两种蛋白为包被抗原均为阳性的血清最多,占到总数的58.5%,前者阳性后者阴性的血清阳性率28%,前者阴性后者阳性的血清阳性率8.75%,二者均为阴性的占到4.75%.为建立区别两种毒株的鉴别诊断间接ELISA试剂盒奠定基础.
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| 关 键 词： | 表达 免疫学 |
Expression of Hypervariable Region in PRRSV Nsp2 Gene and Identification of Its Immunogenicity Characteristics |
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| MA Li-hong,XIAO Yi-hong,LV Jun-hua,WANG Wei-wei,DONG Shi-wei,ZHOU En-min.Expression of Hypervariable Region in PRRSV Nsp2 Gene and Identification of Its Immunogenicity Characteristics[J].Southwest China Journal of Agricultural Sciences,2011,24(1). |
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| Authors: | MA Li-hong XIAO Yi-hong LV Jun-hua WANG Wei-wei DONG Shi-wei ZHOU En-min |
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| Abstract: | Hypervariable region in Nsp2 gene of isolate(TA-12) and ATTC-VR2332 were cloned and subcloned into pET-28a prokaryotic expression system to get recombinant protein after induction of IPTG.The recombinant proteins were identified by SDS-PAGE and Western-blot.BALB/c mice were immunized to analyze their immunogenicity.An indirect ELISA was developed coating with recombinant protein.400 sera were detected by the developed ELISA.The results showed hypervariable region in Nsp2 gene of isolate and ATTC-VR2332 were... |
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| Keywords: | PRRSV Nsp2 ELISA |
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