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目的通过原核表达获得埃博拉病毒科特迪瓦型的重组核蛋白,并将蛋白纯化。方法将合成的埃博拉病毒科特迪瓦型核蛋白基因亚克隆入原核表达质粒pET-28(a)中,构建重组表达质粒pET-28(a)-C-NP,并将重组质粒转化BL21(DE3)感受态细菌,以IPTG诱导蛋白表达,SDS-PAGE分析表达蛋白及表达量,并用利用His-Band Ni+柱进行亲和层析纯化,Western blot和蛋白序列分析鉴定表达蛋白。结果所构建的重组表达质粒pET-28(a)-C-NP序列正确,SDS-PAGE检测到目的蛋白表达,表达蛋白正确。结论成功表达并纯化了埃博拉病毒科特迪瓦型核蛋白,为后续研究奠定了基础。
OBJECTIVE: To obtain the Ebola-type recombinant nucleoprotein of Ebola virus by prokaryotic expression and to purify the protein. Methods The synthetic Ebola virus Cote d’Ivoire nucleoprotein gene was subcloned into the prokaryotic expression plasmid pET-28 (a) to construct the recombinant expression plasmid pET-28 (a) -C-NP. The recombinant plasmid was transformed into BL21 ) Competent cells. IPTG was used to induce the expression of the protein. SDS-PAGE was used to analyze the expressed protein and its expression. The recombinant protein was purified by affinity chromatography using His-Band Ni + column. The expressed protein was identified by Western blot and protein sequence analysis. Results The constructed recombinant plasmid pET-28 (a) -C-NP had the correct sequence. The target protein was detected by SDS-PAGE and the expressed protein was correct. Conclusion The successful expression and purification of Ebola Côte d’Ivoire nucleoprotein laid the foundation for further research.