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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。
The full-length nucleocapsid (N) protein of the SARS coronavirus and the N-terminal or / and C-terminal deletion mutants were expressed by using the E. coli expression system and 39 recombinant proteins were expressed in 15% -30% between. Recombinant proteins were purified by electroelution or metal chelating agent respectively. Western blotting was used to detect the reactivity of the purified protein to convalescent sera of patients with SARS. The results showed that the full-length N protein had the best activity and the rest of the deleted proteins could not reach the same Activity level. This suggests that the integrity of the N protein is necessary for adequate exposure of its predominant epitopes.