Expression, purification, and characterization of SARS coronavirus RNA polymerase
文献类型：期刊论文
| 作者 | Cheng, A; Zhang, W; Xie, YH; Jiang, WH; Arnold, E; Sarafianos, SG; Ding, JP |
| 刊名 | VIROLOGY
 |
| 出版日期 | 2005 |
| 卷号 | 335期号:2页码:165-176 |
| 关键词 | SARS coronavirus RNA-dependent RNA polymerase polymerization |
| 通讯作者 | Ding, JP (reprint author), Chinese Acad Sci, Shanghai Inst Biol Sci, Inst Biochem & Cell Biol, Key Lab Proteom, 320 Yue Yang Rd, Shanghai 200031, Peoples R China.,jpding@sibs.ac.cn |
| 英文摘要 | The RNA-dependent RNA polymerase (RdRp) of SARS coronavirus (SARS-CoV) is essential for viral replication and a potential target for anti-SARS drugs. We report here the cloning, expression, and purification of the N-terminal GST-fused SARS-CoV RdRp and its polymerase catalytic domain in Escherichia coli. During purification, the full-length GST-RdRp was found to cleave into three main fragments: an N-terminal p12 fragment, a middle p30 fragment, and a C-terminal p64 fragment comprising the catalytic domain, presumably due to bacterial proteases. Biochemical assays show that the full-length GST-RdRp has RdRp activity and the p64 and p12 fragments form a complex that exhibits comparable RdRp activity, whereas the GST-p64 protein has no activity, suggesting that the p 12 domain is required for polymerase activity possibly via involvement in template-primer binding. Nonnucleoside HIV-1 RT inhibitors are shown to have no evident inhibitory effect on SARS-CoV RdRp activity. This work provides a basis for biochemical and structural studies of SARS-CoV RdRp and for development of anti-SARS drugs. (c) 2005 Elsevier Inc. All rights reserved. |
| 学科主题 | Virology |
| 类目[WOS] | Virology |
| 关键词[WOS] | ACUTE RESPIRATORY SYNDROME ; MOUSE HEPATITIS-VIRUS ; HIV-1 REVERSE-TRANSCRIPTASE ; DOUBLE-MEMBRANE VESICLES ; REPLICATION COMPLEX ; CRYSTAL-STRUCTURES ; IDENTIFICATION ; INHIBITOR ; PROTEINS ; RESOLUTION |
| 收录类别 | SCI |
| 语种 | 英语 |
| WOS记录号 | WOS:000228761600003 |
| 版本 | 出版稿 |
| 源URL | [http://202.127.25.143/handle/331003/1859]  |
| 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 | Cheng, A,Zhang, W,Xie, YH,et al. Expression, purification, and characterization of SARS coronavirus RNA polymerase[J]. VIROLOGY,2005,335(2):165-176. |
| APA | Cheng, A.,Zhang, W.,Xie, YH.,Jiang, WH.,Arnold, E.,...&Ding, JP.(2005).Expression, purification, and characterization of SARS coronavirus RNA polymerase.VIROLOGY,335(2),165-176. |
| MLA | Cheng, A,et al."Expression, purification, and characterization of SARS coronavirus RNA polymerase".VIROLOGY 335.2(2005):165-176. |
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