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 Weiyu; Yang Xinning; Jin Daozhong; Zhu Xingzu |
| 刊名 | Acta Pharmacologica Sinica
 |
| 出版日期 | 2004 |
| 卷号 | 25期号:8页码:1000 |
| 关键词 | SELECTIVE-INHIBITION ANTIINFLAMMATORY DRUGS SYNTHASE-1 BIOSYNTHESIS COX-2 baculoviruses prostaglandin-endoperoxide synthase non-steroidal anti inflammatory agents prostaglandins drug inhibition |
| ISSN号 | 1671-4083 |
| 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. |
| 语种 | 英语 |
| 源URL | [http://119.78.100.183/handle/2S10ELR8/283752]  |
| 专题 | 中国科学院上海药物研究所 |
| 作者单位 | 中国科学院上海药物研究所 |
| 推荐引用方式 GB/T 7714 | Zhang Weiyu,Yang Xinning,Jin Daozhong,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. |
| APA | Zhang Weiyu,Yang Xinning,Jin Daozhong,&Zhu Xingzu.(2004).Expression and enzyme activity determination of human cyclooxygenase-1 and -2 in a baculovirus-insect cell system.Acta Pharmacologica Sinica,25(8),1000. |
| MLA | Zhang Weiyu,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. |
入库方式： OAI收割
来源：上海药物研究所
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