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• W2078634573 abstract "DNA methyltransferase (MTase) is a kind of important regulatory factor in various biological processes. Current methods to investigate DNA MTase activity are still limited in the sensitivity and/or generality. Therefore, developing methods with high sensitivity and improved generality is needed. Here, we develop a new bioluminescence strategy based on methylation-resistant cleavage and protein expression in vitro to detect DNA MTase activity. In the strategy, Dam MTase was used as a model enzyme and MboI as the methylation-resistant endonuclease, and luciferase reporter DNA (LR–DNA) was used as their action target. Because the completely methylated LR–DNA could be expressed as detectable luciferase, Dam MTase activity was quantified by measuring the luminescence intensity of the expressed luciferase. The assay provides a very low detection limit (0.08 U/ml) as well as a wide linear range (0.2–100 U/ml). Besides, the analysis mode has improved generality and could be extended to the detection of other DNA MTases and the corresponding inhibitor screening." @default.
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• W2078634573 date "2012-04-01" @default.
• W2078634573 modified "2023-10-18" @default.
• W2078634573 title "A bioluminescence assay for DNA methyltransferase activity based on methylation-resistant cleavage" @default.
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• W2078634573 doi "https://doi.org/10.1016/j.ab.2012.01.013" @default.
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