Zhang Xiao, Zhong Xiao-Li, Jiang Wen-Wen, Zeng Si-Hao, Pi Ting, Zheng Xiang-Juan, Li Zhi-Mei
Department of Chemistry, Nanchang University.
School of Nursing of Jiangxi University of Technology.
Anal Sci. 2018;34(8):959-964. doi: 10.2116/analsci.18P080.
DNA methyltransferase (MTase) is related to transcriptional repressor activity in biological functions. It is an essential for cancer diagnosis and therapeutics to detect DNA MTase activity sensitively. Here, a fluorescent system based on polymerase amplification has been developed to detect DNA adenine MTase (Dam) activity sensitively. The amplification is triggered by the probe DNA regions a, which are the primes of a polymerase-induced replicated reaction. They come from methylation and a digestion reaction of DNA S1-S1, including a 5'-GATC-3' sequence recognized by Dam MTase and methylation sensitive restriction endonuclease Dpn I. The intensities of fluorescence are dependent on the Dam MTase activity. The method shows fine sensitivity with a detection limit of 3.2 × 10 U mL and specificity for Dam MTase. In human serum samples, the method has been successfully applied, and it has also been used to screen the inhibitors, which means that the developed method can be a powerful and potential tool for drug development and clinical diagnosis in the future.
DNA甲基转移酶（MTase）在生物学功能中与转录抑制活性相关。灵敏地检测DNA MTase活性对于癌症诊断和治疗至关重要。在此，已开发出一种基于聚合酶扩增的荧光系统，用于灵敏地检测DNA腺嘌呤MTase（Dam）活性。扩增由探针DNA区域a触发，区域a是聚合酶诱导的复制反应的引物。它们来自DNA S1 - S1的甲基化和消化反应，包括Dam MTase识别的5'-GATC-3'序列以及甲基化敏感限制性内切酶Dpn I。荧光强度取决于Dam MTase活性。该方法具有良好的灵敏度，检测限为3.2×10 U/mL，且对Dam MTase具有特异性。在人血清样本中，该方法已成功应用，并且还用于筛选抑制剂，这意味着所开发的方法未来可能成为药物开发和临床诊断的强大且有潜力的工具。
