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马尔堡病毒核蛋白的原核表达及纯化 被引量:1

Prokaryotic Expression and Purification of Nucleoprotein of Marburg Virus
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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。 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.
出处 《中国动物检疫》 CAS 2011年第10期35-37,共3页 China Animal Health Inspection
基金 公益性行业(农业)科研专项经费(200903037-06) 江苏检验检疫局科技计划项目(2011KJ03)
关键词 马尔堡病毒 NP蛋白 原核表达 蛋白纯化 Marburg vires NP protein prokaryotic expression protein purification
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