| 埃博拉病毒苏丹型核蛋白的原核表达及纯化 |
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| 引用本文： | 王雪敏,王皓婷,邱亚峰,史子学,魏建超,邵东华,王志亮,马志永.埃博拉病毒苏丹型核蛋白的原核表达及纯化[J].中国兽医科学,2011(2). |
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| 作者姓名： | 王雪敏 王皓婷 邱亚峰 史子学 魏建超 邵东华 王志亮 马志永 |
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| 作者单位： | 河北工程大学农学院;中国农业科学院上海兽医研究所兽医公共卫生研究室;中国动物卫生与流行病学中心; |
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| 基金项目： | 公益性行业(农业)科研专项(200903037-06) |
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| 摘 要： | 将目的基因NP亚克隆入原核表达质粒pET-28(a)中,构建了重组表达载体pET-28(a)-S-NP,然后用重组质粒转化BL21(DE3)感受态细菌,并利用IPTG诱导蛋白表达。将表达的蛋白利用His-Band Ni+柱进行亲和层析纯化,Western-blot鉴定表达蛋白。结果显示,重组质粒经双酶切后可得到与目的片段长度相同的特异性条带;测序结果显示此特异性条带与参考序列完全匹配,没有突变。纯化后的蛋白经SDS-PAGE和Western-blot鉴定,表达正确。结果表明,成功构建了重组表达质粒,且表达产物正确。
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| 关 键 词： | 埃博拉病毒 核蛋白 原核表达 蛋白纯化 |
Prokaryotic expression and purification of nucleoprotein of Ebola virus Sudan type |
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| WANG Xue-min,WANG Hao-ting,QIU Ya-feng,SHI Zi-xue,WEI Jian-chao,SHAO Dong-hua,WANG Zhi-liang,MA Zhi-yong.Prokaryotic expression and purification of nucleoprotein of Ebola virus Sudan type[J].Veterinary Science in China,2011(2). |
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| Authors: | WANG Xue-min WANG Hao-ting QIU Ya-feng SHI Zi-xue WEI Jian-chao SHAO Dong-hua WANG Zhi-liang MA Zhi-yong |
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| Institution: | WANG Xue-min1,WANG Hao-ting1,2,QIU Ya-feng2,SHI Zi-xue2,WEI Jian-chao2,SHAO Dong-hua2,WANG Zhi-liang3,MA Zhi-yong2(1.College of Agriculture,Hebei University of Engineering,Handan 056021,China,2.Department of Veterinary Public Health,Shanghai Veterinary Research Institute,Chinese Academy of Agricultural Sciences,Shanghai 200241,3.China Animal Health and Epidemiology Center,Qingdao 266114,China) |
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| Abstract: | To express and purify the recombinant protein of the nucleoprotein(NP) of Ebola virus Sudan type,the synthesized NP gene was sub-cloned into plasmid pET-28(a),and the constructed recombinant plasmid pET-28(a)-S-NP was subsequently transformed into Escherichia coli BL21(DE3).The expression of the recombinant plasmid was induced by IPTG and analyzed by SDS-PAGE and Western-blot.The recombinant protein was purified using affinity chromatography.In result,the recombinant protein was expressed from the recombina... |
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| Keywords: | Ebola virus nucleoprotein prokaryotic expression protein purification |
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