In Situ, Real-Time Monitoring of the 3 ' to 5 ' Exonucleases Secreted by Living Cells
文献类型：期刊论文
| 作者 | Su, Xin; Zhu, Xiaocui; Zhang, Chen; Xiao, Xianjin; Zhao, Meiping |
| 刊名 | ANALYTICAL CHEMISTRY
 |
| 出版日期 | 2012-06-05 |
| 卷号 | 84期号:11页码:5059-5065 |
| ISSN号 | 0003-2700 |
| DOI | 10.1021/ac300745f |
| 英文摘要 | Enzymes containing 3'-5' exonuclease activities play vital roles in maintaining genome stability. Though a wide variety of methods have been developed for detection of these enzymes, few of them can be directly applied for in situ and real-time monitoring of the secretion of these active substances by living cells. Taking advantages of the free 3'-end of stacked guanine-quenched photoinduced electron transfer fluorescent probes, here we demonstrate a novel assay capable of in situ and real-time monitoring of the 3'-5' exonucleases secreted by living cells. The detection limit of the new method achieved as low as 0.04 U/mL, allowing direct monitoring of the target enzymes in an extracellular environment without preconcentration steps. False positive signals caused by other nonspecific enzymes were easily ruled out by the use of a control probe with the 3'-end modified with exonuclease-resistant phosphorothioate guanines. Using Alexa Fluor 488 as the fluorophore, the probe is adaptable to a wide range of pH conditions. The approach was successfully applied for in situ, real-time monitoring of the 3'-5' exonucleases secreted by suspension cells of Arabidopsis thaliana. It also holds great potential for in situ and real-time detection of many other DNA end-processing enzymes produced by other types of cells. |
| 语种 | 英语 |
| WOS记录号 | WOS:000304783100060 |
| 出版者 | AMER CHEMICAL SOC |
| 源URL | [http://ir.iccas.ac.cn/handle/121111/49629]  |
| 专题 | 中国科学院化学研究所 |
| 通讯作者 | Zhao, Meiping |
| 作者单位 | Peking Univ, Beijing Natl Lab Mol Sci, MOE Key Lab Bioorgan Chem & Mol Engn, Coll Chem & Mol Engn, Beijing 100871, Peoples R China |
| 推荐引用方式 GB/T 7714 | Su, Xin,Zhu, Xiaocui,Zhang, Chen,et al. In Situ, Real-Time Monitoring of the 3 ' to 5 ' Exonucleases Secreted by Living Cells[J]. ANALYTICAL CHEMISTRY,2012,84(11):5059-5065. |
| APA | Su, Xin,Zhu, Xiaocui,Zhang, Chen,Xiao, Xianjin,&Zhao, Meiping.(2012).In Situ, Real-Time Monitoring of the 3 ' to 5 ' Exonucleases Secreted by Living Cells.ANALYTICAL CHEMISTRY,84(11),5059-5065. |
| MLA | Su, Xin,et al."In Situ, Real-Time Monitoring of the 3 ' to 5 ' Exonucleases Secreted by Living Cells".ANALYTICAL CHEMISTRY 84.11(2012):5059-5065. |
入库方式： OAI收割
来源：化学研究所
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