Electrochemical biosensing method for the detection of DNA methylation and assay of the methyltransferase activity
Electrochemical biosensing method for the detection of DNA methylation and assay of the methyltransferase activity
复制标题
电化学生物传感法检测DNA甲基化和甲基转移酶活性
DOI：
10.1016/j.snb.2012.12.124
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发表时间：
2013-03-01
影响因子：
8.4
通讯作者：
Ai, Shiyun
中科院分区：
文献类型：
--
作者：
Xu, Zhenning;Wang, Mo;Ai, Shiyun
本文介绍了一种以亚甲基蓝（MB）为电化学指示剂检测DNA甲基化和测定DNA甲基转移酶（MTase）活性的电化学方法。DNA杂交体被T.aqI MTase甲基化后，不能被HincII内切酶切割。相反，没有甲基化的双链DNA可以被切割，这将减少嵌入MB的量。因此，基于MB的伏安信号变化，可以确定DNA甲基化状态和MTase活性。利用该方法可以找到甲基化位点。MB的电化学信号随着T. aq I MTase浓度的增加而线性增加，从0.1到100 U/mL。抑制实验表明，非瑟酮对T.aqIMTase有抑制作用，IC_（50）为280 μ M。因此，该方法可用于MTase抑制剂的筛选。(C)2013 Elsevier B.V.保留所有权利。
In this article, we presented an electrochemical method for detection of DNA methylation and assay of DNA methyltransferase (MTase) activity using methylene blue (MB) as electrochemical indicator. After the DNA hybrid was methylated by T.aqI MTase, it could not be cleaved by HincII endonuclease. On the contrary, the double DNA without methylation could be cleaved, which would decrease the amount of intercalated MB. Thus, the DNA methylation status and MTase activity could be determinated based on the voltammetric signal change of MB. The methylation site could be found based on this method. The electrochemical signal of MB increased linearly with increasing T.aqI MTase concentration from 0.1 to 100 U/mL. Moreover, the inhibition investigation demonstrated that fisetin could inhibit the T.aqI MTase activity with the IC50 value of 280 mu M. Therefore, the screening of the inhibitors of MTase could be accomplished using the novel method. (C) 2013 Elsevier B.V. All rights reserved.