Rapid and sensitive detection of DNA polymerase fidelity by singly   labeled smart fluorescent probes

doi:10.1016/j.bios.2010.08.073

CORC > 北京大学 > 化学与分子工程学院

	Rapid and sensitive detection of DNA polymerase fidelity by singly labeled smart fluorescent probes
	Song, Chen ; Zhang, Chen ; Zhao, Meiping
刊名	生物传感器与生物电子学
	2011
关键词	DNA polymerase fidelity Singly labeled smart fluorescent probe Real-time monitoring Mispair extension Proofreading activity REPLICATION FIDELITY MISPAIR EXTENSION BIOCHEMICAL BASIS ESCHERICHIA-COLI KLENOW FRAGMENT WILD-TYPE BASE 2-AMINOPURINE BACTERIOPHAGE-T4 NUCLEOTIDES
DOI	10.1016/j.bios.2010.08.073
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.; Biophysics; Biotechnology & Applied Microbiology; Chemistry, Analytical; Electrochemistry; Nanoscience & Nanotechnology; SCI(E); PubMed; 7; ARTICLE; 5; 2699-2702; 26
语种	英语
内容类型	期刊论文
源URL	[http://ir.pku.edu.cn/handle/20.500.11897/150000]
专题	化学与分子工程学院
推荐引用方式 GB/T 7714	Song, Chen,Zhang, Chen,Zhao, Meiping. Rapid and sensitive detection of DNA polymerase fidelity by singly labeled smart fluorescent probes[J]. 生物传感器与生物电子学,2011.
APA	Song, Chen,Zhang, Chen,&Zhao, Meiping.(2011).Rapid and sensitive detection of DNA polymerase fidelity by singly labeled smart fluorescent probes.生物传感器与生物电子学.
MLA	Song, Chen,et al."Rapid and sensitive detection of DNA polymerase fidelity by singly labeled smart fluorescent probes".生物传感器与生物电子学 (2011).