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

Abstract

Herein, a novel fluorescent method for detection of T4 polynucleotide kinase (T4 PNK) based on the FRET between dye labeled DNA and polydopamine nanotubes (PDANTs). PDANTs can bind and quench fluorescent dye-labeled single-stranded DNA (ssDNA) probes, while it has less affinity toward double-stranded DNA (dsDNA). With the T4 PNK and ATP, λ exonuclease initiated the hydrolysis of the 5-phosphoryl termini of dsDNA and release ssDNA, which can be strongly adsorbed on PDANTs and the fluorescence intensity decreased. On basis of this, we have successfully established a proposed method for detecting T4 PNK. The method displayed a detection limit of 0.01 U mL-1 with a linear range from 0.01 to 50 U mL-1. In addition, we investigate the effect of several inhibitors including (NH4)2SO4 and NaCl. The method provides a platform for T4 PNK detection and evaluate inhibitors.

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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, Accepted Manuscript

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

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

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