期刊
BIOSENSORS & BIOELECTRONICS
卷 32, 期 1, 页码 213-218出版社
ELSEVIER ADVANCED TECHNOLOGY
DOI: 10.1016/j.bios.2011.12.012
关键词
Electrochemical; T4 polynucleotide kinase; Single wall carbon nanotubes; Titanium ion
类别
资金
- Natural Science Foundation of China [90606003, 21175039, 21190044]
- International Science & Technology Cooperation Program of China [2010DFB30300]
- Hunan Provincial Natural Science Foundation
- Hunan Provincial Science and Technology Plan of China [10JJ7002, 2011FJ2001]
- Hunan Provincial Innovation Foundation For Postgraduate
A novel electrochemical strategy for monitoring the activity and inhibition of 14 polynucleotide kinase (PNK) is developed by use of titanium ion (Ti4+) mediated signal transition coupled with signal amplification of single wall carbon nanotubes (SWCNTs). In this method, a DNA containing 5'-hydroxyl group is self-assembled onto the gold electrode and used as substrate for PNK. The biofunctionalized SWCNTs with anchor DNA and ferrocene are chosen as the signal indicator by virtue of the intrinsic 5'-phosphate end of anchor DNA and the high loading of ferrocene for electrochemical signal generation and amplification. The 5'-hydroxyl group of the substrate DNA on the electrode is phosphorylated by T4 PNK in the presence of ATP, and the resulting 5'-phosphoryl end product can be linked with the signal indicator by Ti4+. The redox ferrocene group on the SWCNTs is grafted to the electrode and generates the electrochemical signal, the intensity of which is proportional to the activity of 14 PNK. This assay can measure activity of T4 PNK down to 0.01 U mL(-1). The developed method is a potentially useful tool in researching the interactions between proteins and nucleic acids and provides a diversified platform for a kinase activity assay. (C) 2011 Elsevier B.V. All rights reserved.
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