Cell-free bioassay for measurement of dioxins based on fluorescence enhancement of fluorescein isothiocyanate-labeled DNA probe
文献类型：期刊论文
| 作者 | You, Fan; Zhou, Ya-Feng; Zhang, Xian-En; Huang, Zhi; Bi, Li-Jun; Zhang, Zhi-Ping; Wen, Ji-Kai; Chen, Yuan-Yuan; Jiang, Gui-Bin; Zheng, Ming-Hui |
| 刊名 | ANALYTICAL CHEMISTRY
 |
| 出版日期 | 2006 |
| 卷号 | 78期号:20页码:7138-7144 |
| 英文摘要 | This study aims to develop a rapid and sensitive cell-free bioassay of dioxins. It is known that dioxin ligand can bind heterodimeric aryl hydrocarbon receptor (AhR) and triggers the formation of the complex of dioxin-AhR, AhR nuclear translocator (ARNT), and dioxin-responsive element (DRE) region of the DNA. The hypothesis of the proposed method is that if FITC were labeled at the DRE sequence, its fluorescence intensity would be enhanced when the complex forms because the interaction interface of the binding components (AhR, ARNT, and DRE) creates a rather hydrophobic condition that is in favor of FITC emission. Effects of modification site of FITC on the DNA probes on binding efficiency between the complex components and fluorescence emission enhancement were evaluated by surface plasmon resonance and fluorescence analysis, respectively. Results showed that the labeling site at the second base at the 5' end apart from the core region (5'-TNGCGTG-3') of DRE did not obviously interfere with the binding between the DNA probe and dioxin-AhR/ARNT hybrid but presented significant fluorescence emission enhancement. 2,3,7,8-Tetrachlorodibenzo-p-dioxin (TCDD) was used as the typical toxin in this study. The method had a linear range of 1- 100 pM, with detection limit of 0.1 pM (0.64 fg/assay) and coefficient of variation of 5.6% (n = 10, 50 pM TCDD in transformed cytosol). The whole detection cycle was similar to 4 h. The method was also used to estimate the toxic equivalents (TEQ) of 1,2,3,7,8-pentachlorodibenzo-p-dioxin (PeCDD) and 1,2,3,4,7,8-hexachlorodibenzo-p-dioxin ( HxCDD). Measurement of TEQs of the mixture of TCDD, PeCDD, and HxCDD were highly consistent with the predicted data. The average recovery using fly ash extract was similar to 93%. |
| WOS记录号 | WOS:000241219800009 |
| 源URL | [http://ir.rcees.ac.cn/handle/311016/22840]  |
| 专题 | 生态环境研究中心_环境化学与生态毒理学国家重点实验室 |
| 推荐引用方式 GB/T 7714 | You, Fan,Zhou, Ya-Feng,Zhang, Xian-En,et al. Cell-free bioassay for measurement of dioxins based on fluorescence enhancement of fluorescein isothiocyanate-labeled DNA probe[J]. ANALYTICAL CHEMISTRY,2006,78(20):7138-7144. |
| APA | You, Fan.,Zhou, Ya-Feng.,Zhang, Xian-En.,Huang, Zhi.,Bi, Li-Jun.,...&Zheng, Ming-Hui.(2006).Cell-free bioassay for measurement of dioxins based on fluorescence enhancement of fluorescein isothiocyanate-labeled DNA probe.ANALYTICAL CHEMISTRY,78(20),7138-7144. |
| MLA | You, Fan,et al."Cell-free bioassay for measurement of dioxins based on fluorescence enhancement of fluorescein isothiocyanate-labeled DNA probe".ANALYTICAL CHEMISTRY 78.20(2006):7138-7144. |
入库方式： OAI收割
来源：生态环境研究中心
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