• 文献标题:   Graphene oxide-hairpin probe nanocomposite as a homogeneous assay platform for DNA base excision repair screening
  • 文献类型:   Article
  • 作  者:   ZHOU DM, XI Q, LIANG MF, CHEN CH, TANG LJ, JIANG JH
  • 作者关键词:   graphene oxide, uracildna glycosylase, hairpin probe
  • 出版物名称:   BIOSENSORS BIOELECTRONICS
  • ISSN:   0956-5663
  • 通讯作者地址:   Hunan Univ
  • 被引频次:   48
  • DOI:   10.1016/j.bios.2012.08.053
  • 出版年:   2013

▎ 摘  要

Uracil-DNA glycosylase (UDG) as one of the most important base excision repair enzymes plays a crucial role in protecting the genome from endogenous DNA damage and sustaining the genome integrity. Quantitative activity analysis of UDG is a central challenge and of fundamental importance in bioanalysis. Here, we proposed a novel biosensor constituted by adsorbing a fluorophore-labeled hairpin probe onto the surface of graphene oxide (GO) as a homogeneous assay platform for sensitive UDG activity assay. Active UDG could excise the uracil base in the hairpin probe, and further hydrolysis of the leaving abasic site gave rise to high fluorescence. Thus, it provided a convenient approach for UDG activity quantification. Because of the unique ability of GO in universal fluorescence quenching, a low background fluorescence signal can be obtained for the efficient fluorescence resonant energy transfer from the fluorophore-labeled on the hairpin probe to GO sheet. A quite wide dynamic range from 0.0017 U/mL to 0.8 U/mL was achieved for UDG assay and the detection limit was estimated to be 0.0008 U/mL. The results indicated that this strategy offers a simple, cost-effective, highly sensitive and selective homogeneous detection platform for UDG activity assay related biochemical studies. (C) 2012 Elsevier B.V. All rights reserved.