期刊
BIOSENSORS & BIOELECTRONICS
卷 34, 期 1, 页码 100-105出版社
ELSEVIER ADVANCED TECHNOLOGY
DOI: 10.1016/j.bios.2012.01.024
关键词
DNAzyme; Molecular beacon; Exonuclease III; DNA detection; Endonuclease
类别
资金
- National Natural Science Foundation of China [20935003, 21075116]
- 973 project [2011CB911002, 2010CB933603]
A novel G-quadruplex DNAzyme molecular beacon (G-DNAzymeMB) strategy is developed for assays of target DNA and restriction endonuclease. The detection system consists of G-DNAzymeMB strand and a blocker DNA by using the fluorescence of 2',7'-dichlorodihydrofluorescein diacetate (H(2)DCFDA) catalyzed by G-DNAzymeMB as a signal reporter. G-DNAzymeMB exhibits peroxidase activity in its free hairpin structure, and forms a catalytically inactive hybrid when hybridized with blocker DNA. Upon displacement of blocker DNA by target DNA or cleavage by restriction endonuclease. G-DNAzymeMB is released and two lateral portions of G-DNAzymeMB form a G-quadruplex structure, resulting in the recovery of catalytic activity which acts as a cofactor to catalyze H2O2-mediated oxidation of H(2)DCFDA. For DNA detection system, exonuclease III (Exo III)-catalyzed amplification strategy is introduced to improve the sensitivity and target DNA could be detected as low as 0.1 pM. With respect to restriction endonuclease detection system, 0.1 U/mL EcoRI endonuclease could be detected and this method could be easily transported to other restriction endonuclease analysis by simply changing the recognition sequence. These results demonstrate that the proposed G-DNAzymeMB strategy could be used as a label-free, simple, sensitive and cost-effective approach in analysis of target DNA and restriction endonuclease. (C) 2012 Elsevier B.V. All rights reserved.
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