ICCAS OpenIR
Rapid and sensitive detection of DNA polymerase fidelity by singly labeled smart fluorescent probes
Song, Chen; Zhang, Chen; Zhao, Meiping
2011-01-15
Source PublicationBIOSENSORS & BIOELECTRONICS
ISSN0956-5663
Volume26Issue:5Pages:2699-2702
AbstractWe 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.
KeywordDna Polymerase Fidelity Singly Labeled Smart Fluorescent Probe Real-time Monitoring Mispair Extension Proofreading Activity
DOI10.1016/j.bios.2010.08.073
Indexed BySCI
Language英语
WOS IDWOS:000286904400149
PublisherELSEVIER ADVANCED TECHNOLOGY
Citation statistics
Cited Times:17[WOS]   [WOS Record]     [Related Records in WOS]
Document Type期刊论文
Identifierhttp://ir.iccas.ac.cn/handle/121111/72462
Collection中国科学院化学研究所
Corresponding AuthorZhao, Meiping
AffiliationPeking Univ, Coll Chem & Mol Engn, Key Lab Bioorgan Chem & Mol Engn, Beijing Natl Lab Mol Sci,Minist Educ, Beijing 100871, Peoples R China
Recommended Citation
GB/T 7714
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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