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• W2016149021 abstract "• DNA demethylase activity was assayed by electrochemical method. • Double enzyme system of DNA demethylase and BstUI endonuclease was applied. • The detection is based on alkaline phosphatase catalytic signal amplification. • This method showed high detection selectivity and good sensitivity. A novel electrochemical method is developed for detection of DNA demethylation and assay of DNA demethylase activity. This method is constructed by hybridizing the probe with biotin tagged hemi-methylated complementary DNA and further capturing streptavidin tagged alkaline phosphatase (SA-ALP) to catalyze the hydrolysis reaction of p -nitrophenyl phosphate. The hydrolysate of p -nitrophenol (PNP) is then used as electrochemical probe for detecting DNA demethylation and assaying the activity of DNA demethylase. Demethylation of target DNA initiates a degradation reaction of the double-stranded DNA (dsDNA) by restriction endonuclease of BstUI. It makes the failed immobilization of ALP, resulting in a decreased electrochemical oxidation signal of PNP. Through the change of this electrochemical signal, the DNA demethylation is identified and the activity of DNA demethylase is analyzed with low detection limit of 1.3 ng mL −1 . This method shows the advantages of simple operation, cheap and miniaturized instrument, high selectivity. Thus, it provides a useful platform for detecting DNA demethylation, analyzing demethylase activity and screening inhibited drug." @default.
• W2016149021 created "2016-06-24" @default.
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• W2016149021 date "2014-08-01" @default.
• W2016149021 modified "2023-10-18" @default.
• W2016149021 title "Enzyme-based electrochemical biosensor for sensitive detection of DNA demethylation and the activity of DNA demethylase" @default.
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• W2016149021 doi "https://doi.org/10.1016/j.aca.2014.06.020" @default.
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