SARS病毒核衣壳蛋白在大肠杆菌中的表达及纯化  

Expression of SARS-CoV Neucleocapsid Protein in E.coli and Purification of Expressed Product

作　　者：李洋[1] 王宣军[2] 张秀霞[2] 吉海滨[2] 王志武[2] 方勇[1] 盛军[2] 

机构地区：[1]吉林大学生命科学学院,长春130021 [2]长春生物制品研究所,长春130062

出　　处：《中国生物制品学杂志》2005年第6期465-466,469,共3页Chinese Journal of Biologicals

摘　　要：目的在大肠杆菌中高效表达SARS核衣壳蛋白。方法将已合成的SARS-CoV N蛋白基因片段,克隆至pUC-18载体,与pET28质粒连接,转化大肠杆菌,进行SARS-CoV核衣壳融合蛋白表达。用SARS患者恢复期血清进行表达产物鉴定。用色谱方法进行表达产物的层析纯化。结果SARS病毒N基因在大肠杆菌中获得了高效表达,表达量占总蛋白的30%以上;经三步纯化后纯度达90%以上。结论已获得了SARS核衣壳蛋白样品。Objective To highly express SARS-CoV neucleocapsid protein in E. coll. Methods Insert the synthetic SARS-CoV N protein gene into vector pUC-18,ligate to plasmid pET28 and transform to E. coli for expression. Identify the expressed product by the sera of patients with SARS during recovery phase. Purify the expressed product by chromatography. Results SARS-CoV N gene was highly expressed in E. coll. The expressed product contained more than 30% of total somatic protein. The purity of expressed product after three steps of chromatography reached more than 90%. Conclusion SARS neucleocapsid protein for further study was obtained.

关 键 词：SARS病毒 非典型性肺炎 核衣壳蛋白 大肠杆菌 基因表达 纯化方式 

分 类 号：R563.1[医药卫生—呼吸系统] R378.21[医药卫生—内科学]