High-level expression and single-step purification of human tryptophanyl-tRNA synthetase
文献类型：期刊论文
| 作者 | Xu, F; Jia, J; Jin, YX; Wang, DTP |
| 刊名 | PROTEIN EXPRESSION AND PURIFICATION
 |
| 出版日期 | 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 |
| 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. |
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