A sensitive electrochemical assay for T4 polynucleotide kinase activity based on titanium dioxide nanotubes and a rolling circle amplification strategy

Zhang, Yanli, Fang, Xiang, Zhu, Zhenyu, Lai, Yanqiong, Xu, Chunli, Pang, Pengfei, Wang, Hongbin, Yang, Chun, Barrow, Colin J. and Yang, Wenrong 2018, A sensitive electrochemical assay for T4 polynucleotide kinase activity based on titanium dioxide nanotubes and a rolling circle amplification strategy, RSC advances, vol. 8, no. 67, pp. 38436-38444, doi: 10.1039/c8ra07745b.

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Title A sensitive electrochemical assay for T4 polynucleotide kinase activity based on titanium dioxide nanotubes and a rolling circle amplification strategy
Author(s) Zhang, Yanli
Fang, Xiang
Zhu, Zhenyu
Lai, Yanqiong
Xu, Chunli
Pang, Pengfei
Wang, Hongbin
Yang, Chun
Barrow, Colin J.ORCID iD for Barrow, Colin J. orcid.org/0000-0002-2153-7267
Yang, WenrongORCID iD for Yang, Wenrong orcid.org/0000-0001-8815-1951
Journal name RSC advances
Volume number 8
Issue number 67
Start page 38436
End page 38444
Total pages 9
Publisher Royal Society of Chemistry
Place of publication Cambridge, Eng.
Publication date 2018-11
ISSN 2046-2069
Keyword(s) Science & Technology
Physical Sciences
Chemistry, Multidisciplinary
Chemistry
LAMBDA EXONUCLEASE
KINASE/PHOSPHATASE ACTIVITY
ENZYMATIC PHOSPHORYLATION
DEOXYRIBONUCLEIC-ACID
COLORIMETRIC ASSAY
RIBONUCLEIC-ACID
BIOSENSOR
REPAIR
NANOPARTICLES
PLATFORM
Summary © 2018 The Royal Society of Chemistry. An ultrasensitive electrochemical biosensor was developed for detection of T4 polynucleotide kinase (T4 PNK) activity based on titanium dioxide nanotubes (TiO 2 NTs) and a rolling circle amplification (RCA) strategy. In this study, the immobilized T4 PNK substrate probe with a 5′ terminus hydroxyl was phosphorylated by T4 PNK in the presence of adenosine triphosphate (ATP), and the resulting 5-phosphoryl can be linked with the TiO 2 NTs and further conjugated with the phosphate-labeled primer. RCA was initiated by adding circular template, phi29 DNA polymerase and deoxyribonucleoside 5-triphosphate mixture (dNTPs). Biotin-labeled probes are chosen as a signal indicator by strong biotin-streptavidin interaction and the high loading of horseradish peroxidase-streptavidin (HRP-SA) for electrochemical signal generation and amplification. A dual-signaling amplification strategy has been established, which exhibited an excellent performance with a wide linear range from 0.0001-15 U mL -1 and a low detection limit of 0.00003 U mL -1 for T4 PNK detection. The inhibition effect of (NH 4 ) 2 SO 4 on the activity of T4 PNK is also evaluated. This new dual-signaling electrochemical biosensor can be used for the detection of the activity and inhibition of other nucleic acid enzymes.
Language eng
DOI 10.1039/c8ra07745b
HERDC Research category C1 Refereed article in a scholarly journal
ERA Research output type C Journal article
Copyright notice ©2018, The Royal Society of Chemistry
Persistent URL http://hdl.handle.net/10536/DRO/DU:30115996

Document type: Journal Article
Collections: School of Life and Environmental Sciences
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