| 乙型脑炎病毒SA14-14-2株NS1和E蛋白的原核表达及其免疫原性 |
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| 引用本文： | 徐宏山,俞永新,贾丽丽,黄莺,董关木,俞炜源.乙型脑炎病毒SA14-14-2株NS1和E蛋白的原核表达及其免疫原性[J].粉末涂料与涂装,2010,23(2). |
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| 作者姓名： | 徐宏山 俞永新 贾丽丽 黄莺 董关木 俞炜源 |
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| 作者单位： | 徐宏山,俞永新,贾丽丽,董关木(中国药品生物制品检定所疫苗一室,北京,100050);黄莺,俞炜源(军事医学科学院生物工程研究所蛋白质工程室,北京,100071) |
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| 摘 要： | 目的原核表达乙型脑炎病毒(JEV)NS1和E蛋白,并初步探讨其免疫原性。方法采用RT-PCR法扩增JEVSA14-14-2株NS1和E蛋白基因片段,分别克隆入原核表达载体pET-30a(+)和pET-32a(+),构建重组表达质粒pET-30NS1和pET-32Et,转化大肠杆菌BL21(DE3),IPTG诱导表达,SDS-PAGE分析目的蛋白的表达形式及表达水平。两种蛋白经亲和层析纯化后,Western blot鉴定其反应原性。通过小鼠主动和被动保护力试验及豚鼠病毒血症试验,检测两种蛋白的免疫原性。结果酶切及测序证明重组表达质粒构建正确;表达的重组NS1和E蛋白的相对分子质量分别约为40000和47000,均以包涵体形式表达;纯化的重组NS1和E蛋白的浓度分别为160.7和380μg/ml,均具有良好的反应原性;小鼠主动和被动保护力试验表明两种蛋白均有保护活性;豚鼠病毒血症试验显示,NS1蛋白免疫豚鼠后能明显抑制病毒血症的产生。结论已成功表达并纯化了JEVSA14-14-2株NS1和E蛋白,两种蛋白均有较好的免疫保护活性。
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| 关 键 词： | 乙型脑炎病毒 SA14-14-2株 NS1蛋白 E蛋白 原核表达 免疫原性 |
Prokaryotic Expression and Immunogenicity of NS1 and E Proteins of Japanese Encephalitis Virus SA14-14-2 Strain |
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| XU Hong-shan,YU Yong-xin,JIA Li-li,et al.Prokaryotic Expression and Immunogenicity of NS1 and E Proteins of Japanese Encephalitis Virus SA14-14-2 Strain[J].Chinese Journal of Biologicals,2010,23(2). |
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| Authors: | XU Hong-shan YU Yong-xin JIA Li-li |
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| Affiliation: | XU Hong-shan,YU Yong-xin,JIA Li-li,et al(National Institute for the Control of Pharmaceutical , Biological Products,Beijing 100050,China) |
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| Abstract: | Objective To express the NS1 and E proteins of Japanese encephalitis virus(JEV)SA14-14-2 strain in prokaryotic cells and study its immunogenicity. Methods The gene fragments encoding NS1 and E proteins of JEV SA14-14-2 strain were amplified by RT-PCR and cloned into prokaryotic expression vectors pET-30a(+)and pET-32a(+)respectively. The constructed recombinant plasmids pET-30NS1 and pET-32Et were transformed to E. coli BL21(DE3)respectively for expression under induction of IPTG. The expressed product was ... |
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| Keywords: | Japanese encephalitis virus (JEV) SA14-14-2 strain NS1 protein E protein Prokaryotic expression Immunogenicity |
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