Journal
JOURNAL OF PHARMACEUTICAL AND BIOMEDICAL ANALYSIS
Volume 219, Issue -, Pages -Publisher
ELSEVIER
DOI: 10.1016/j.jpba.2022.114935
Keywords
Methylation; Strand displacement amplification; Fluorescence detection; Methyltransferase activity
Categories
Funding
- National Natural Science Foundations of China [21565011, 22064008]
- Guangxi Natural Science Foundation [2017GXNSFAA198340]
- China Post-doctoral Science Foundation [2017M612873, GLUTQD2002047-1]
- Guangxi Key Laboratory of Environmental Pollution Control Theory and Technology
- Guangxi Key Laboratory of Electrochemicaland Magnetochemical Functional Materials
- Guangxi Colleges and Universities Key Laboratory of Food Safety and Detection
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A rapid and sensitive fluorescence sensing strategy for the detection of Dam MTase activity was developed based on methylation-blocked enzymatic recycling amplification. This method has a lower detection limit and shorter assay time compared with previously reported similar methodologies.
DNA methylation catalyzed by DNA adenine methylation methyltransferase (Dam MTase) is strongly connected with a variety of biological processes, hence, monitoring Dam MTase activity is of great importance. Here, we developed a rapid and sensitive fluorescence sensing strategy for the detection of Dam MTase activity based on methylation-blocked enzymatic recycling amplification. In this fluorescence sensing system, Dam MTase-induced methylation blocked the subsequent reactions. In contrast, in the absence of Dam MTase, the unmethylated probe initiated the cascade strand displacement amplification for significant signal amplification. Under optimized conditions, this method has a lower detection limit of 0.67 U/mL and a shorter assay time (90 min) compared with previously reported similar methodologies.
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