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| 題名: | Establishment of a cell-free bioassay for detecting dioxin-like compounds |
| 作者: | 吳沛翊;Wang, Bo-Jeng;Wu, Pei-Yi;Lu, Yi-Chien;Chang, Chi-Hao;Lin, Yueh-Chien;Tsai, Tzu-Ching;Hsu, Ming-Ching;Lee, Hsinyu |
| 貢獻者: | 生醫理工學院生命科學系 |
| 關鍵詞: | AhR;Arnt;Aryl Hydrocarbon Receptor Nuclear Translocator - genetics;Aryl Hydrocarbon Receptor Nuclear Translocator - metabolism;bioassay;Biological Assay - methods;Blotting, Western;dioxin;Dioxins - analysis;Environmental Pollutants - analysis;Fluorescent Dyes - metabolism;HEK293 Cells;Humans;Limit of Detection;Luciferases, Renilla - genetics;Luciferases, Renilla - metabolism;Receptors, Aryl Hydrocarbon - genetics;Receptors, Aryl Hydrocarbon - metabolism;Recombinant Fusion Proteins - genetics;Recombinant Fusion Proteins - metabolism;Renilla;Transfection |
| 日期: | 2013-07-01 |
| 上傳時間: | 2026-04-23 11:12:37 (UTC+8) |
| 出版者: | Informa Healthcare;England: Informa Healthcare USA, Inc |
| 摘要: | 摘要： Abstract Dioxins are byproducts from incomplete combustion processes and belong to a group of mostly toxic chemicals known as persistent organic pollutants, and 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) is considered to be the most toxic species of all dioxin-like compounds. Analytical chemical processes are employed to determine the specific dioxin content in environmental samples. However, cost-ineffectiveness and excess time consumption limit their routine utilization. The aryl hydrocarbon receptor (AhR) is the major TCDD receptor. Upon binding to dioxin, the AhR dissociates from Hsp90 and other cofactors. TCDD-bound AhR subsequently translocates to the nucleus and interacts with the AhR nuclear translocator (Arnt) to induce signal transduction. Here, we describe a highly sensitive and cost-effective alternative assay based on detecting stability of bioluminescence signals. We generated cells that stably co-express Renilla luciferase tagged-AhR (AhR-RL), Ah receptor-interacting protein (AIP), p23 and yellow fluorescent protein-tagged Arnt (Arnt-YFP) (AAPA cells) for detection of dioxin-like compounds. Treatment with 3-methylcholanthrene (3MC), AhR agonist, enhanced the interaction between AhR and Arnt and avoided proteosomal degradation. In addition, treatment with 3MC or TCDD stabilized Renilla luminescence from AhR-RL of AAPA cell-free extracts in a concentration-dependent manner. The TCDD detection limit in this cell-free system was as low as 10−18 M. These results highlight the potential of AAPAA cell-free extracts to detect dioxin-like pollutants.
其他題名： Toxicol Mech Methods
出版者： England: Informa Healthcare USA, Inc
出版日期： 2013-07-01
出處： Toxicology Mechanisms and Methods, 2013-07, Vol.23 (6), p.464-470
資源來源： EBSCOhost Academic Search Premier
版權： 2013 Informa Healthcare USA, Inc. All rights reserved: reproduction in whole or part not permitted 2013
識別號： ISSN: 1537-6516
識別號： ISSN: 1537-6524
識別號： EISSN: 1537-6524
識別號： DOI: 10.3109/15376516.2013.781254
識別號： PMID: 23477502 |
| 顯示於類別: | [生命科學系] 期刊論文
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