SARS冠状病毒核衣壳(N)蛋白不同区域的原核表达

Prokaryotic Expression of Different Regions of SARS-CoV Nucleocapsid Protein

利用大肠杆菌表达系统对SARS冠状病毒的核衣壳(N)蛋白全长及N末端或/和C末端缺失突变体进行了表达,共表达了39个重组蛋白,表达量在15%～30%之间。分别利用电洗脱或金属鳌合介质纯化重组蛋白,用蛋白印迹实验检测纯化蛋白对SARS病人恢复期血清的反应性,结果发现全长N蛋白活性最好,其余的末端缺失蛋白均无法达到同一活性水平。由此说明N蛋白的完整性对于其优势表位的充分暴露是必要的。

Nucleocapsid(N)protein of the severe acute respiratory syndrome associated coronavirus(SARS-CoV) , and its N or/and C terminal truncated mutants were expressed in E. coli .The yield levels of these thirty-nine recombinant proteins were from 15% to 30% .Recombinant proteins were purified by metal chelated affinity chromatography or by electro-elution. Western blot was used to detect the reactivity of these proteins with the convalescent sera of SARS patients. The results showed that the reactivity of the full length N protein is the best,which suggests that the integrity of N protein is important for exposure of the prominent epitopes.