DNA
巴米
化学
电化学
纳米团簇
纳米线
信号(编程语言)
铜
纳米技术
生物化学
分析化学(期刊)
材料科学
色谱法
电极
计算机科学
限制性酶
物理化学
有机化学
程序设计语言
作者
Yufang Hu,Qingqing Zhang,Lihua Xu,Jiao Wang,Jiajia Rao,Zhiyong Guo,Sui Wang
标识
DOI:10.1007/s00216-017-0623-0
摘要
Electrochemical methods allow fast and inexpensive analysis of enzymatic activity. Here, a simple and yet efficient signal-on electrochemical assay for sensitive, label-free detection of DNA-related enzyme activity was established on the basis of terminal deoxynucleotidyl transferase (TdT)-mediated extension strategy. TdT, which is a template-independent DNA polymerase, can catalyze the sequential addition of deoxythymidine triphosphate (dTTP) at the 3'-OH terminus of single-stranded DNA (ssDNA); then, the TdT-yield T-rich DNA nanowires can be employed as the synthetic template of copper nanoclusters (CuNCs). Grown DNA nanowires-templated CuNCs (noted as DNA-CuNCs) were attached onto graphene oxide (GO) surface and exhibited unique electrocatalytic activity to H2O2 reduction. Under optimal conditions, the proposed biosensor was utilized for quantitatively monitoring TdT activity, with the observed LOD of 0.1 U/mL. It also displayed high selectivity to TdT with excellent stability, and offered a facile, convenient electrochemical method for TdT-relevant inhibitors screening. Moreover, the proposed sensor was successfully used for BamHI activity detection, in which a new 3'-OH terminal was exposed by the digestion of a phosphate group. Ultimately, it has good prospects in DNA-related enzyme-based biochemical studies, disease diagnosis, and drug discovery. Graphical Abstract Extraordinary TdT-generated DNA-CuNCs are synthesized and act as a novel electrochemical sensing platform for sensitive detection of TdT and BamHI activity in biological environments.
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