4.7 Article

A colorimetric assay of DNA methyltransferase activity based on the keypad lock of duplex DNA modified meso-SiO2@Fe3O4

期刊

ANALYTICA CHIMICA ACTA
卷 920, 期 -, 页码 80-85

出版社

ELSEVIER SCIENCE BV
DOI: 10.1016/j.aca.2016.03.028

关键词

DNA methyltransferase; meso-SiO2@Fe3O4; Paclitaxel; Colorimetric assay

资金

  1. National Natural Science Foundation of China [21375079, 21105056]
  2. Project of Development of Science and Technology of Shandong Province, China [2013GZX20109]
  3. Natural Science Foundation of Shandong province China [ZR2014BQ029]

向作者/读者索取更多资源

Abnormal level of DNA methyltransferase (MTase) - mediated DNA methylation is closely related with cancer and bacterial diseases. Herein, a novel strategy based on the keypad lock of duplex DNA modified meso-SiO2@Fe3O4 was developed for colorimetric assay of Dam MTase activity. When the Dam MTase was introduced, the duplex DNA can be methylated at a palindrome sequence of 5'-GATC-3' and cleaved by the methylation-sensitive restriction endonuclease Dpn I. Due to the instability of the newly formed DNA fragment, the hybrid will separated into a single-stranded DNA. Then the keypad lock will open, and the catalytic reaction of TMB and H2O2 can be initiated through the pores of meso-SiO2@Fe3O4, and a high color signal can be clearly observed by the naked eye. Contrarily, without Dam MTase, the catalytic reaction will not be initiated, and result no color signal. The proposed method exhibited a wide dynamic range with a low detection limit of 0.73 U/mL. Additionally, this way can be performed in human serum with satisfying recovery. And the inhibition of Dam MTase can also be well demonstrated by using paclitaxel as a model. Therefore, the designed way not only provides a platform for monitoring Dam MTase activity, but also useful for further application in disease diagnosis and drug discovery. (C) 2016 Elsevier B.V. All rights reserved.

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