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Microchimica Acta

, 186:100 | Cite as

A hairpin-type DNA probe for direct colorimetric detection of endonuclease activity and inhibition based on the deaggregation of gold nanoparticles

  • Fuming Sang
  • Guoliang Li
  • Jun Li
  • Jianxin Pan
  • Zhizhou Zhang
  • Xue Zhang
Original Paper
  • 28 Downloads

Abstract

A method is described for the determination of the activity of endonuclease. It based on the deaggregation of gold nanoparticles (AuNPs) aggregated by the action of poly(diallyldimethylammonium chloride) (PDDA). A single-stranded DNA (ssDNA) is released after enzymatic cleavage catalyzed by endonuclease. The released fragments bind electrostatically to PDDA and inhibit the PDDA-induced aggregation of AuNPs. This is accompanied by a color change from blue to red and a decrease in the absorption ratio (A630/A520). Under the optimal conditions, this ratio increases linearly in the 0.001 to 1 U·μL−1 EcoRI endonuclease activity range. The detection limit is of 2 × 10−4 U·μL−1 which is much better or at least comparable to previous reports. The method is deemed to have wide scope in that it may be used to study other endonuclease activity (such as BamHI) by simply changing the specific recognition site of the hairpin-like DNA probe. The assay may also be employed to screening for inhibitors of EcoRI endonuclease.

Graphical abstract

Schematic presentation of the colorimetric assay based on the deaggregation of AuNPs for the detection of endonuclease activity. A single-stranded sequence (ssDNA) is released by the EcoRI cleavage, which electrostatically binds to PDDA and inhibits the PDDA-induced aggregation of AuNPs accompanying with a color change from blue to red.

Keywords

EcoRI Gold nanoparticle Visual test Deaggregation Endonuclease inhibitor 

Notes

Acknowledgments

This work was funded by the Natural Science Foundation of China (NSFC) (No. 21407035), Shandong Provincial Natural Science Foundation (ZR2014BM021), Technology and Development Program of Weihai (2014DXGJ15), HIT-NSRIF (2011101).

Compliance with ethical standards

All procedures were in compliance with ethical strandards.

Supplementary material

604_2018_3164_MOESM1_ESM.doc (2.2 mb)
ESM 1 (DOC 2299 kb)

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Copyright information

© Springer-Verlag GmbH Austria, part of Springer Nature 2019

Authors and Affiliations

  • Fuming Sang
    • 1
  • Guoliang Li
    • 1
  • Jun Li
    • 1
  • Jianxin Pan
    • 1
  • Zhizhou Zhang
    • 1
  • Xue Zhang
    • 1
  1. 1.School of Marine Science and TechnologyHarbin Institute of TechnologyWeihaiPeople’s Republic of China

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