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A novel fluorescence method for the highly sensitive detection of T4 polynucleotide kinase based on polydopamine nanotubes

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Abstract

Herein, a novel fluorescence method is reported for the detection of T4 polynucleotide kinase (T4 PNK) based on a FRET between dye-labeled DNA and polydopamine nanotubes (PDANTs). PDANTs can bind and quench the fluorescent dye-labeled single-stranded DNA (ssDNA) probes, whereas they have low affinity towards double-stranded DNA (dsDNA). In the presence of T4 PNK and ATP, λ exonuclease initiated the hydrolysis of the 5-phosphoryl termini of dsDNA and released ssDNA, which could strongly adsorb on the PDANTs and decrease the fluorescence intensity; on the basis of this concept, we successfully established a method for the detection of T4 PNK. The method displayed the detection limit of 0.01 U mL−1 with a linear range from 0.01 to 50 U mL−1. In addition, we investigated the effect of several inhibitors including Na2HPO4, (NH4)2SO4 and NaCl. The method provides a platform for the detection of T4 PNK and evaluation of inhibitors.

Graphical abstract: A novel fluorescence method for the highly sensitive detection of T4 polynucleotide kinase based on polydopamine nanotubes

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Publication details

The article was received on 24 Aug 2019, accepted on 29 Sep 2019 and first published on 30 Sep 2019


Article type: Paper
DOI: 10.1039/C9NJ04381K
New J. Chem., 2019, Advance Article

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    A novel fluorescence method for the highly sensitive detection of T4 polynucleotide kinase based on polydopamine nanotubes

    X. Shen, J. Ge, J. Chen, Y. Shen, H. Meng, Z. Li and L. Qu, New J. Chem., 2019, Advance Article , DOI: 10.1039/C9NJ04381K

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