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| 马尔堡病毒核蛋白的原核表达及纯化 | |
| 引用本文： | 王皓婷,王水明,史子学,邵东华,魏建超,刘学辉,王志亮,王雪敏,马志永.马尔堡病毒核蛋白的原核表达及纯化[J].中国动物检疫,2011,28(10):35-37. |
| 作者姓名： | 王皓婷 王水明 史子学 邵东华 魏建超 刘学辉 王志亮 王雪敏 马志永 |
| 作者单位： | 1. 中国农业科学院上海兽医研究所兽医公共卫生研究室,上海,200241 2. 南京出入境检验检疫局,江苏南京,211106 3. 中国动物卫生与流行病学中心,山东青岛,266114 4. 河北工程大学农学院,河北邯郸,056001 |
| 基金项目： | 公益性行业(农业)科研专项经费，江苏检验检疫局科教计划项目 |
| 摘 要： | 目的以原核表达的方式获得了马尔堡病毒的核蛋白,并纯化。方法将马尔堡病毒核蛋白基因亚克隆到原核表达质粒pET-28(a)中,构建重组表达质粒pET-28(a)-M-NP,并将重组质粒用热休克方法转化BL21(DE3)感受态细菌,用IPTG诱导蛋白表达后,以SDS-PAGE分析表达形式及表达量,并利用His-Band Ni+柱纯化。结果成功构建重组表达质粒pET-28(a)-M-NP,以1.0mmol/L终浓度的IPTG在37℃诱导表达5h后,破菌沉淀中有目的蛋白表达。结论成功表达并纯化了马尔堡病毒的核蛋白,为后续研究奠定了基础。 |
| 关 键 词： | 马尔堡病毒 NP蛋白 原核表达 蛋白纯化 |
Prokaryotic Expression and Purification of Nucleoprotein of Marburg Virus |
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| Wang Haoting,Wang Shuiming,Shi Zixue,Shao Donghu,Wei Jianchao,Liu Xuehui,Wang Zhiliang,Wang Xuemin,Ma Zhiyong.Prokaryotic Expression and Purification of Nucleoprotein of Marburg Virus[J].China Journal Of Animal Quarantine,2011,28(10):35-37. | |
| Authors: | Wang Haoting Wang Shuiming Shi Zixue Shao Donghu Wei Jianchao Liu Xuehui Wang Zhiliang Wang Xuemin Ma Zhiyong |
| Institution: | Wang Haoting1,2,Wang Shuiming3,ShiZixue1,Shao Donghua1,Wei Jianchao1,Liu Xuehui3,Wang Zhiliang4,Wang Xuemin2,Ma Zhiyong1 (1.Department of Veterinary Public Health,Shanghai Veterinary Research Institute,Chinese Academy of Agricultural Sciences,Shanghai,200241,China,2.College of Agriculture,Hebei university of engineering,Handan,Hebei,056001,3.Nanjing Entry-Exit Inspection and Quarantine Bureau,Nanjing,Jiangsu,211106,4.China Animal Health and Epidemiology Center,Qingdao,266114,China) |
| Abstract: | Objective To obtain recombinant protein of the nucleoprotein （NP） of Marburg virus using prokaryotic expression system. Synthesized NP gene was sub-cloned to generate pET-28 （a） -M-NP recombinant plasmid, and then transformed into E.eoli BL21 （DE3） competent bacteria.The expression of NP recombinant protein was induced by IPTG and analyzed by SDA-PAGE. The expressed protein was purified using His-Band Ni ＋ affinity chromatography and identified by Western blot. Results The recombinant expression plasmid pET-28 （a） -M-NP was obtained and the recombinant NP protein was expressed and purified. Conclusion The NP recombinant protein was expressed and characterized. |
| Keywords: | Marburg vires NP protein prokaryotic expression protein purification |
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