甲基转移酶
DNA甲基转移酶
DNA
化学
荧光
CpG站点
分子生物学
检出限
DNA甲基化
甲基化
滚动圆复制
核酸内切酶
生物物理学
生物化学
生物
DNA复制
基因
色谱法
物理
基因表达
量子力学
作者
Qilin Wen,Dandan Li,Guidan Huang,Huai Xi,Hongcheng Pan,Lianming Zhang,Ziyuan Li,Xiaofen Xiao,Wenyuan Zhu
出处
期刊:Analyst
[The Royal Society of Chemistry]
日期:2022-01-01
卷期号:147 (22): 4980-4985
被引量:3
摘要
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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