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Rapid and sensitive detection of DNA polymerase fidelity by singly labeled smart fluorescent probes
Song, Chen; Zhang, Chen; Zhao, Meiping
关键词Dna Polymerase Fidelity Singly Labeled Smart Fluorescent Probe Real-time Monitoring Mispair Extension Proofreading Activity
刊名BIOSENSORS & BIOELECTRONICS
2011-01-15
DOI10.1016/j.bios.2010.08.073
26期:5页:2699-2702
收录类别SCI
文章类型Article
WOS标题词Science & Technology ; Life Sciences & Biomedicine ; Physical Sciences
类目[WOS]Biophysics ; Biotechnology & Applied Microbiology ; Chemistry, Analytical ; Electrochemistry ; Nanoscience & Nanotechnology
研究领域[WOS]Biophysics ; Biotechnology & Applied Microbiology ; Chemistry ; Electrochemistry ; Science & Technology - Other Topics
关键词[WOS]REPLICATION FIDELITY ; MISPAIR EXTENSION ; BIOCHEMICAL BASIS ; ESCHERICHIA-COLI ; KLENOW FRAGMENT ; WILD-TYPE ; BASE ; 2-AMINOPURINE ; BACTERIOPHAGE-T4 ; NUCLEOTIDES
英文摘要We report here a novel approach to monitor the DNA polymerase fidelity in detailed steps, including mispair extension, mispair formation and 3' -> 5' proofreading. The method is based on the photo-induced electron transfer between the natural base guanine and the labeled fluorophore. The G:T mispair extension catalyzed by the exonuclease-deficient Klenow fragment DNA polymerase (KF exo(-)) was easily detected and the effect of the nearest neighbor base pair on the mispair extension rate was clearly observed. More importantly, kinetics of the G:T, G:A and G:G mispair formation and extension under single turnover conditions were measured by continuous fluorescence-based assay for the first time. The probes also showed their applicability to discriminate the 3' -> 5' proofreading activity of different exonuclease-proficient DNA polymerases. The presented method may greatly simplify the screening and characterization procedures of the increasing number of polymerases that are thought to be potential targets for drug design and cancer treatment. It will also provide important information for deep understanding of the polymerase fidelity mechanism. (C) 2010 Elsevier B.V. All rights reserved.
语种英语
WOS记录号WOS:000286904400149
引用统计
文献类型期刊论文
条目标识符http://cas-ir.dicp.ac.cn/handle/321008/142479
专题中国科学院大连化学物理研究所
作者单位Peking Univ, Coll Chem & Mol Engn, Key Lab Bioorgan Chem & Mol Engn, Beijing Natl Lab Mol Sci,Minist Educ, Beijing 100871, Peoples R China
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Song, Chen,Zhang, Chen,Zhao, Meiping. Rapid and sensitive detection of DNA polymerase fidelity by singly labeled smart fluorescent probes[J]. BIOSENSORS & BIOELECTRONICS,2011,26(5):2699-2702.
APA Song, Chen,Zhang, Chen,&Zhao, Meiping.(2011).Rapid and sensitive detection of DNA polymerase fidelity by singly labeled smart fluorescent probes.BIOSENSORS & BIOELECTRONICS,26(5),2699-2702.
MLA Song, Chen,et al."Rapid and sensitive detection of DNA polymerase fidelity by singly labeled smart fluorescent probes".BIOSENSORS & BIOELECTRONICS 26.5(2011):2699-2702.
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