PCR Incorporation of Polyoxometalate Modified Deoxynucleotide Triphosphates and Their Application in Molecular Electrochemical Sensing of Yersinia pestis

Chemistry. 2017 Aug 4;23(44):10597-10603. doi: 10.1002/chem.201701295. Epub 2017 Jul 18.

Abstract

Redox-labeled nucleotides are of increasing interest for the fabrication of next generation molecular tools and should meet requirements of being thermally stable, sensitive, and compatible with polymerase-mediated incorporation while also being electrochemically discriminable. The synthesis and characterization of Keggin and Dawson polyoxometalate-deoxynucleotide (POM-dNTP) bioconjugates linked through 7-deaza-modified purines is described. The modified POM-dNTPs were used for polymerase-based amplification of a DNA sequence specific for Yersinia pestis and the amplified DNA detected using an electrochemical DNA sensor. This highlights the potential of polyoxometalates as thermally stable, sensitive and polymerase-compatible redox labels for exploitation in bioanalytical applications.

Keywords: biochemistry; electrochemistry; oligonucleotides; polymerase chain reaction; polyoxometalates.

MeSH terms

  • DNA, Bacterial / chemistry*
  • DNA, Bacterial / metabolism
  • Electrochemical Techniques*
  • Electrodes
  • Electrophoresis, Gel, Pulsed-Field
  • Gold / chemistry
  • Nucleotides / chemistry*
  • Polymerase Chain Reaction
  • Tungsten Compounds / chemistry*
  • Yersinia pestis / genetics*
  • Yersinia pestis / isolation & purification

Substances

  • DNA, Bacterial
  • Nucleotides
  • Tungsten Compounds
  • polyoxometalate I
  • Gold