Molecular cloning, expression, purification, and mass spectrometric characterization of 3C-like protease of SARS coronavirus

	Molecular cloning, expression, purification, and mass spectrometric characterization of 3C-like protease of SARS coronavirus
	Sun, HF; Luo, HB; Yu, CY; Sun, T; Chen, J; Peng, SY; Qin, J; Shen, JH; Yang, YM; Xie, YH
刊名	PROTEIN EXPRESSION AND PURIFICATION
	2003
卷号	32 期号:2 页码:302-308
关键词	Severe Acute Respiratory Syndrome (SARS) SARS coronavirus 3C like protease molecular cloning expression purification mass spectrometric characterization
通讯作者	Jiang, HL (reprint author), Chinese Acad Sci, Shanghai Inst Biol Sci, Shanghai Inst Mat Med, State Key Lab Drug Res,Drug Discovery & Design Ct, Shanghai 201203, Peoples R China.,
英文摘要	Severe acute respiratory syndrome (SARS) is an acute respiratory illness, which has broken out in China. It has been known that SARS coronavirus (SARS_CoV) is a novel human coronavirus and is responsible for SARS infection. Belonging to one of the major proteins associated with SARS_CoV, SARS 3C-like protease (SARS_3CL(pro)) functions as a cysteine protease engaging in the proteolytic cleavage of the viral precursor polyprotein to a series of functional proteins required for coronavirus replication and is considered as an appealing target for designing anti-SARS agents. To facilitate the studies regarding the functions and structures of SARS_3CL(pro), in this report the synthetic genes encoding 3CL(pro), of SARS_CoV were assembled, and the plasmid was constructed using pQE30 as vector and expressed in Escherichia coli M15 cells. The highly yielded (similar to15 mg/L) expressed protease was purified by use of NTA-Ni2+ affinity chromatography and FPLC system, and its sequence was determined by LC/MS with the residue coverage of 46.4%. (C) 2003 Elsevier Inc. All rights reserved.
学科主题	Biochemistry & Molecular Biology; Biotechnology & Applied Microbiology
类目[WOS]	Biochemical Research Methods ; Biochemistry & Molecular Biology ; Biotechnology & Applied Microbiology
关键词[WOS]	ACUTE RESPIRATORY SYNDROME ; VIRUS-ENCODED PROTEINASES ; RNA-POLYMERASE ; SEQUENCE ; IDENTIFICATION ; POLYPROTEIN ; KILOBASES ; DESIGN ; GENE-1 ; DOMAIN
收录类别	SCI
语种	英语
WOS记录号	WOS:000187393600017
内容类型	期刊论文
版本	出版稿
源URL	[http://202.127.25.143/handle/331003/2242]
专题	上海生化细胞研究所_上海生科院生化细胞研究所
推荐引用方式 GB/T 7714	Sun, HF,Luo, HB,Yu, CY,et al. Molecular cloning, expression, purification, and mass spectrometric characterization of 3C-like protease of SARS coronavirus[J]. PROTEIN EXPRESSION AND PURIFICATION,2003,32(2):302-308.
APA	Sun, HF.,Luo, HB.,Yu, CY.,Sun, T.,Chen, J.,...&Jiang, HL.(2003).Molecular cloning, expression, purification, and mass spectrometric characterization of 3C-like protease of SARS coronavirus.PROTEIN EXPRESSION AND PURIFICATION,32(2),302-308.
MLA	Sun, HF,et al."Molecular cloning, expression, purification, and mass spectrometric characterization of 3C-like protease of SARS coronavirus".PROTEIN EXPRESSION AND PURIFICATION 32.2(2003):302-308.