• 文献标题:   A facile graphene oxide-based DNA polymerase assay
  • 文献类型:   Article
  • 作  者:   XU FZ, SHI H, HE XX, WANG KM, YE XS, YAN LA, WEI SY
  • 作者关键词:  
  • 出版物名称:   ANALYST
  • ISSN:   0003-2654 EI 1364-5528
  • 通讯作者地址:   Hunan Univ
  • 被引频次:   15
  • DOI:   10.1039/c2an35585j
  • 出版年:   2012

▎ 摘  要

The DNA polymerase assay is fundamental for related molecular biology investigations and drug screenings, however, the commonly used radioactive method is laborious and restricted. Herein, we report a novel, simple and cost-effective fluorometric DNA polymerase detection method by utilizing graphene oxide (GO) as a signal switch. In this strategy, in the absence of DNA polymerase, the fluorophore-labeled template ssDNA could be strongly adsorbed and almost entirely quenched by GO. However, as DNA polymerase exists, the polymerized dsDNA product might lead to a much lower quenching efficiency after addition of GO due to the much weaker interaction of dsDNA with GO than ssDNA, thus resulting in a much higher fluorescence signal detected. As proof of concept, the quantitative DNA polymerase activity assay was performed using the Klenow fragment exo(-) (KF-) as a model. It was confirmed that, after optimization of detection conditions, KF- activity could be sensitively detected through facile fluorescence measurements, with a detection limit of 0.05 U mL(-1) and a good linear correlation between 0.05-2.5 U mL(-1) (R-2 - 0.9928). In addition, this GO-based method was further inspected to evaluate the inhibitive behaviors of several drugs toward KF- activity, the result of which firmly demonstrated its potential application in polymerization-targeted drug screening.