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题名: Development of a High-Throughput Screening Platform for DNA 3 '-Phosphatases and Their Inhibitors Based on a Universal Molecular Beacon and Quantitative Real-time PCR
作者: Song, Chen1;  Zhang, Chen1;  Zhao, Mei-Ping1
关键词: DNA ;  fluorescent probes ;  high-throughput screening ;  inhibitors ;  polymerase chain reaction
刊名: CHEMISTRY-AN ASIAN JOURNAL
发表日期: 2010
DOI: 10.1002/asia.200900661
卷: 5, 期:5, 页:1146-1151
收录类别: SCI
文章类型: Article
WOS标题词: Science & Technology ;  Physical Sciences
类目[WOS]: Chemistry, Multidisciplinary
研究领域[WOS]: Chemistry
英文摘要: DNA 3'-phosphatases play a unique role in the repair of strand breaks induced by DNA damaging agents, such as ionizing radiation or oxidative stress. In this paper, we present an efficient detection system for rapid screening of DNA 3'-phosphatases and their inhibitors. A unique template substrate has been designed to hybridize with the universal molecular beacon (U-MB), and the detection process is carried out in a quantitative real-time PCR. The method is successfully applied to monitor the activity and kinetics of two typical 3'-phosphatases, that is, T4 polynucleotide kinase phosphatase (PNKP) and calf intestinal alkaline phosphatase (CIP). The inhibition effect of heparin on T4 PNKP and theophylline on CIP is also quantitatively characterized. The proposed method is demonstrated to be very useful for sensitive, high-throughput, and precise measurement of various 3'-phosphatases and their inhibitors.
关键词[WOS]: T4 POLYNUCLEOTIDE KINASE ;  POLYMERASE-CHAIN-REACTION ;  SINGLE-STRAND BREAKS ;  ALKALINE-PHOSPHATASE ;  3-PHOSPHATASE ACTIVITY ;  SUBSTRATE-SPECIFICITY ;  ACID-PHOSPHATASE ;  RAT-LIVER ;  REPAIR ;  ENZYME
语种: 英语
WOS记录号: WOS:000277724800016
Citation statistics: 
内容类型: 期刊论文
URI标识: http://cas-ir.dicp.ac.cn/handle/321008/141926
Appears in Collections:中国科学院大连化学物理研究所_期刊论文

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作者单位: 1.Peking Univ, Beijing Natl Lab Mol Sci, Key Lab Bioorgan Chem & Mol Engn, Minist Educ,Coll Chem & Mol Engn, Beijing 100871, Peoples R China

Recommended Citation:
Song, Chen,Zhang, Chen,Zhao, Mei-Ping. Development of a High-Throughput Screening Platform for DNA 3 '-Phosphatases and Their Inhibitors Based on a Universal Molecular Beacon and Quantitative Real-time PCR[J]. CHEMISTRY-AN ASIAN JOURNAL,2010,5(5):1146-1151.
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