Journal
ANALYST
Volume 147, Issue 22, Pages 4980-4985Publisher
ROYAL SOC CHEMISTRY
DOI: 10.1039/d2an01302a
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Funding
- National Natural Science Foundations of China [21565011]
- Guangxi Natural Science Foundation [2015GXNSFAA139020]
- China Postdoctoral Science Foundation [2017M612873, GLUTQD2002047-1]
- Guangxi Key Laboratory of Environmental Pollution Control Theory and Technology
- Guangxi Colleges and Universities Key Laboratory of Food Safety and Detection
- Guangxi Key Laboratory of Electrochemical and Magnetochemical Functional Materials
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In this study, a dual amplification fluorescence strategy was proposed for the ultrasensitive detection of DNA adenine methylation methyltransferase (Dam MTase) activity. The method showed a low detection limit, good selectivity, and can be extended to the detection of other types of DNA MTases.
DNA methyltransferase (MTase) is an important regulatory enzyme in various biological processes. However, current methods for investigating MTase activity are still limited in terms of sensitivity and/or generality. Herein, we proposed a dual amplification fluorescence strategy for the ultrasensitive detection of DNA adenine methylation methyltransferase (Dam MTase) activity based on strand displacement amplification (SDA) coupled with rolling circle amplification (RCA). In this study, the hairpin probe could not be cleaved by Nt.AlwI nicking endonuclease (Nt.AlwI) in the presence of Dam MTase, and the subsequent SDA-RCA reaction was blocked, resulting in a weak fluorescence signal. Moreover, the blocking effect was more pronounced at a higher concentration of Dam MTase. This assay provides a very low detection limit (down to 0.0067 U ml(-1)), as well as good selectivity against other types of MTases (e.g., CpG methyltransferase (M.SssI MTase)). In addition, the analytical mode improves the generality and can be extended to the detection of other types of DNA MTases.
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