High-level expression and single-step purification of human tryptophanyl-tRNA synthetase

	High-level expression and single-step purification of human tryptophanyl-tRNA synthetase
	Xu, F; Jia, J; Jin, YX; Wang, DTP
刊名	PROTEIN EXPRESSION AND PURIFICATION
	2001
卷号	23 期号:2 页码:296-300
通讯作者	Jin, YX (reprint author), Chinese Acad Sci, Shanghai Inst Life Sci, Inst Biochem & Cell Biol, State Key Lab Mol Biol, 320 Yue Yang Rd, Shanghai 200031, Peoples R China.,
英文摘要	Human trpS gene was cloned into the expression vector pET-24a(+) to yield pET-24a(+)-HTrpRS, which could direct the synthesis of a mammalian derived protein in Escherichia coli BL21-CodonPlus(DE3)-RIL. The vector allows overproduction and single-step purification of His(6)-tagged human tryptophanyl-tRNA synthetase by the facilitation of metal (Ni2+) chelate affinity chromatography. The expression level of human TrpRS was about 40% of total cell proteins after isopropyl beta -D-thiogalactoside induction. The overproduced human TrpRS-His(6) could be purified to homogeneity within 2 h and about 24 mg purified enzyme could be obtained from 400 ml cell culture. The His(6) tag at C terminus had little effect on the binding ability of its substrates. (C) 2001 Academic Press.
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]	TRANSFER-RNA-SYNTHETASE ; IDENTITY ELEMENTS ; BACILLUS-SUBTILIS ; ESCHERICHIA-COLI ; RECOGNITION ; PROTEINS
收录类别	SCI
语种	英语
WOS记录号	WOS:000171949600011
内容类型	期刊论文
版本	出版稿
源URL	[http://202.127.25.143/handle/331003/2592]
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	Xu, F,Jia, J,Jin, YX,et al. High-level expression and single-step purification of human tryptophanyl-tRNA synthetase[J]. PROTEIN EXPRESSION AND PURIFICATION,2001,23(2):296-300.
APA	Xu, F,Jia, J,Jin, YX,&Wang, DTP.(2001).High-level expression and single-step purification of human tryptophanyl-tRNA synthetase.PROTEIN EXPRESSION AND PURIFICATION,23(2),296-300.
MLA	Xu, F,et al."High-level expression and single-step purification of human tryptophanyl-tRNA synthetase".PROTEIN EXPRESSION AND PURIFICATION 23.2(2001):296-300.