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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
Song, Chen; Zhang, Chen; Zhao, Mei-Ping
关键词Dna Fluorescent Probes High-throughput Screening Inhibitors Polymerase Chain Reaction
刊名CHEMISTRY-AN ASIAN JOURNAL
2010
DOI10.1002/asia.200900661
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
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
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.
语种英语
WOS记录号WOS:000277724800016
引用统计
文献类型期刊论文
条目标识符http://cas-ir.dicp.ac.cn/handle/321008/141926
专题中国科学院大连化学物理研究所
作者单位Peking Univ, Beijing Natl Lab Mol Sci, Key Lab Bioorgan Chem & Mol Engn, Minist Educ,Coll Chem & Mol Engn, Beijing 100871, Peoples R China
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GB/T 7714
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.
APA Song, Chen,Zhang, Chen,&Zhao, Mei-Ping.(2010).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.CHEMISTRY-AN ASIAN JOURNAL,5(5),1146-1151.
MLA Song, Chen,et al."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".CHEMISTRY-AN ASIAN JOURNAL 5.5(2010):1146-1151.
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