军事医学2015，Vol.39Issue(12)：919-922,4.DOI:10.7644/j.issn.1674-9960.2015.12.008
MERS冠状病毒核衣壳蛋白原核表达和纯化
Expression and purification of nucleocapsid protein of MERS coronavirus in E.coli
摘要
Abstract
Objective To construct a prokaryotic expression vector pET-22b+with Middle East respiratory syndrome ( MERS) coronavirus nuclocapsid protein( NP) gene and to express and purify N protein.Methods N gene amplified by PCR was inserted into the prokaryotic expression vector pET-22b+.Recombinant plasmid was confirmed using DNA elec-trophoresis and sequencing.NP was expressed in E.coli BL21(DE3) by IPTG induced and purified by cation exchange chromatography using the AKTA purification system.Results The NP gene sequence was proved to be correct by sequen-cing and the protein was expressed in both soluble and insoluble forms in E.coli BL21 ( DE3 ) after IPTG induction.The purity and concentration of recombinant protein was improved obviously by cation exchange chromatography and enrich-ment.Conclusion Recombiant NP of high purity and concentration is purified and will facilitate NP functional research.关键词
冠状病毒属/MERS-CoVN/原核表达纯化/阳离子交换层析Key words
coronavirus/MERS-CoV N/prokaryotic expression and purification/cation exchange chromatography分类
医药卫生引用本文复制引用
付洋波,胡勇,黄成成,白圆圆,邱立红,曹诚,高婷..MERS冠状病毒核衣壳蛋白原核表达和纯化[J].军事医学,2015,39(12):919-922,4.基金项目
国家973计划资助项目(2012C13518902) （2012C13518902）
