| 乙型脑炎病毒NS1蛋白的可溶性表达与抗原性分析 |
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| 引用本文： | 刘立科,陈振师,任玉红,步志高,华荣虹.乙型脑炎病毒NS1蛋白的可溶性表达与抗原性分析[J].黑龙江畜牧兽医,2012(1):17-19,157. |
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| 作者姓名： | 刘立科 陈振师 任玉红 步志高 华荣虹 |
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| 作者单位： | 河北农业大学动物科技学院;中国农业科学院哈尔滨兽医研究所兽医生物技术国家重点实验室 |
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| 基金项目： | 国家自然科学基金项目(30700027);国家公益性行业科研专项项目(200803015) |
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| 摘 要： | 为获得可溶性表达的乙型脑炎病毒(JEV)NS1蛋白,将编码JEV SA14-14-2株NS1蛋白基因克隆至原核表达载体pMAL-c5X中,构建重组融合表达质粒pc5X-NS1;用重组质粒转化宿主菌ER2523后,经IPTG诱导得到可溶性的融合蛋白MBP-NS1;优化诱导表达条件,于28℃、0.3 mmol/L IPTG诱导4 h;最后融合蛋白经直链淀粉树脂柱纯化。结果表明:重组融合蛋白MBP-NS1具有良好的抗原性和特异性。
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| 关 键 词： | 乙型脑炎病毒 NS1蛋白 原核表达 |
Soluble expression and antigenic analysis of NS1 protein of Japanese encephalitis virus |
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| LIU Li-ke,CHEN Zhen-shi,REN Yu-hong,BU Zhi-gao,HUA Rong-hong.Soluble expression and antigenic analysis of NS1 protein of Japanese encephalitis virus[J].Heilongjiang Animal Science And veterinary Medicine,2012(1):17-19,157. |
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| Authors: | LIU Li-ke CHEN Zhen-shi REN Yu-hong BU Zhi-gao HUA Rong-hong |
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| Institution: | 1.College of Animal Science and Technology,Hebei Agricultural University,Baoding 071001,China; 2.State Key Laboratory of Veterinary Biotechnology,Harbin Veterinary Research Institute,Chinese Academy of Agricultural Sciences,Harbin 150001,China) |
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| Abstract: | To obtain the expression of NS1 protein of Japanese encephalitis virus in a soluble form,the NS1 gene of JEV strain SA14-14-2 was cloned into the expression vector pMAL-c5X,then the recombinant plasmid pc5X-NS1 was confirmed by sequencing.After the recombinant plasmid was transformed into host bacteria ER2523,the fusion protein MBP-NS1 was expressed in the soluble form and purified by induced by isopropyl beta-D-thiogalactopyranoside(IPTG),then the expression conditions were further optimized.Western-blot and indirect ELISA analysis showed that the recombinant fusion protein MBP-NS1 could be specific reactive with antibodies against JEV.This work can provide an important basis for further study on NS1 protein of Japanese encephalitis virus. |
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| Keywords: | Japanese encephalitis virus NS1 protein prokaryotic expression |
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