马尔堡病毒核蛋白的原核表达及纯化

Prokaryotic Expression and Purification of Nucleoprotein of Marburg Virus

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.