期刊
BIOSENSORS & BIOELECTRONICS
卷 86, 期 -, 页码 927-932出版社
ELSEVIER ADVANCED TECHNOLOGY
DOI: 10.1016/j.bios.2016.07.103
关键词
DNA methylation; Methyltransferase; Electroanalysis; DNA superstructure; Restriction endonuclease
类别
资金
- NSFC [21273117, 21375063, 21335004]
- Program for Outstanding Innovation Research Team of Universities in Jiangsu Province
- Priority Academic Program Development of Jiangsu Higher Education Institutions
- Jiangsu Government
DNA methyltransferase (MTase) activity is highly correlated with the occurrence and development of cancer. This work reports a superstructure-based electrochemical assay for signal-amplified detection of DNA MTase activity using M.SssI as an example. First, low-density coverage of DNA duplexes. on the surface of the gold electrode was achieved by immobilized mercaptohexanol, followed by immobilization of DNA duplexes. The duplex can be cleaved by BstUI endonuclease in the absence of DNA superstructures. However, the cleavage is blocked after the DNA is methylated by M.SssI. The DNA superstructures are formed with the addition of helper DNA. By using an electroactive complex, RuHex, which can bind to DNA double strands, the activity of M.SssI can be quantitatively detected by differential pulse voltammetry. Due to the high site-specific cleavage by BstUI and signal amplification by the DNA superstructure, the biosensor can achieve ultrasensitive detection of DNA MTase activity down to 0.025 U/mL. The method can be used for evaluation and screening of the inhibitors of MTase, and thus has potential in the discovery of methylation-related anticancer drugs. (C) 2016 Elsevier B.V. All rights reserved.
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