DICP OpenIR
Rapid and sensitive detection of DNA polymerase fidelity by singly labeled smart fluorescent probes
Song, Chen; Zhang, Chen; Zhao, Meiping
KeywordDna Polymerase Fidelity Singly Labeled Smart Fluorescent Probe Real-time Monitoring Mispair Extension Proofreading Activity
Source PublicationBIOSENSORS & BIOELECTRONICS
2011-01-15
DOI10.1016/j.bios.2010.08.073
Volume26Issue:5Pages:2699-2702
Indexed BySCI
SubtypeArticle
WOS HeadingsScience & Technology ; Life Sciences & Biomedicine ; Physical Sciences
WOS SubjectBiophysics ; Biotechnology & Applied Microbiology ; Chemistry, Analytical ; Electrochemistry ; Nanoscience & Nanotechnology
WOS Research AreaBiophysics ; Biotechnology & Applied Microbiology ; Chemistry ; Electrochemistry ; Science & Technology - Other Topics
WOS KeywordREPLICATION FIDELITY ; MISPAIR EXTENSION ; BIOCHEMICAL BASIS ; ESCHERICHIA-COLI ; KLENOW FRAGMENT ; WILD-TYPE ; BASE ; 2-AMINOPURINE ; BACTERIOPHAGE-T4 ; NUCLEOTIDES
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.
Language英语
WOS IDWOS:000286904400149
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Document Type期刊论文
Identifierhttp://cas-ir.dicp.ac.cn/handle/321008/142479
Collection中国科学院大连化学物理研究所
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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