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

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.