Expression and enzyme activity determination of human cyclooxygenase-1 and-2 in a baculovirus-insect cell system
文献类型：期刊论文
| 作者 | Zhang, WY; Yang, XN; Jin, DZ; Zhu, XZ |
| 刊名 | ACTA PHARMACOLOGICA SINICA
 |
| 出版日期 | 2004-08 |
| 卷号 | 25期号:8页码:1000-1006 |
| 关键词 | baculoviruses prostaglandin-endoperoxide synthase non-steroidal anti inflammatory agents prostaglandins drug inhibition |
| ISSN号 | 1671-4083 |
| 文献子类 | Article |
| 英文摘要 | AIM: To develop an in vitro intact cell-based assay for screening selective cyclooxygenase inhibitors. METHODS: Human cyclooxygenase-1 (hCOX-1) and cyclooxygenase-2 (hCOX-2) genes were cloned from human monocyte cell line THP-1 cells and expressed in Spodoptera frugiperda (sf9) insect cell line by Bac-to-Bac baculovirus expression systems. Infected sf9 cells were harvested 24 h post-infection (hpi), and distributed to a 24-well plate, preincubated with various nonsteroidal anti-inflammatory drugs, and challenged with 10 mmol/L arachidonic acid; the cyclooxygenase activity was assessed indirectly by prostaglandin E(2)-specific radioimmunoassay. RESULTS: Polymerase chain reaction detection demonstrated that hCOX-1 and hCOX-2 were transposed to the bacmid. Western blot analysis showed that infected sf9 cells could express hCOX-1 and hCOX-2 proteins. Radioimmunoassay demonstrated that both recombinant proteins functioned well in sf9 cells. CONCLUSION: Human cyclooxygenase-1 and cyclooxygenase-2 were successfully expressed in sf9 insect cell line. It can be utilized for the identification of potent and selective inhibitors of hCOX-1 and/or hCOX-2. |
| WOS关键词 | SELECTIVE-INHIBITION ; ANTIINFLAMMATORY DRUGS ; SYNTHASE-1 ; BIOSYNTHESIS ; COX-2 |
| WOS研究方向 | Chemistry ; Pharmacology & Pharmacy |
| 语种 | 英语 |
| CSCD记录号 | CSCD:1845122 |
| WOS记录号 | WOS:000223240900005 |
| 出版者 | SHANGHAI INST MATERIA MEDICA |
| 源URL | [http://119.78.100.183/handle/2S10ELR8/274038]  |
| 专题 | 药理学第二研究室 |
| 通讯作者 | Zhu, XZ |
| 作者单位 | Chinese Acad Sci, Shanghai Inst Biol Sci, Shanghai Inst Mat Med, Shanghai 201203, Peoples R China |
| 推荐引用方式 GB/T 7714 | Zhang, WY,Yang, XN,Jin, DZ,et al. Expression and enzyme activity determination of human cyclooxygenase-1 and-2 in a baculovirus-insect cell system[J]. ACTA PHARMACOLOGICA SINICA,2004,25(8):1000-1006. |
| APA | Zhang, WY,Yang, XN,Jin, DZ,&Zhu, XZ.(2004).Expression and enzyme activity determination of human cyclooxygenase-1 and-2 in a baculovirus-insect cell system.ACTA PHARMACOLOGICA SINICA,25(8),1000-1006. |
| MLA | Zhang, WY,et al."Expression and enzyme activity determination of human cyclooxygenase-1 and-2 in a baculovirus-insect cell system".ACTA PHARMACOLOGICA SINICA 25.8(2004):1000-1006. |
入库方式： OAI收割
来源：上海药物研究所
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