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Analytical and Bioanalytical Chemistry

, Volume 406, Issue 15, pp 3755–3762 | Cite as

Specific enrichment of prokaryotic DNA using a recombinant DNA-binding protein

  • Natalia SandetskayaEmail author
  • Andreas Naumann
  • Katharina Hennig
  • Dirk Kuhlmeier
Research Paper

Abstract

Targeted enrichment of DNA is often necessary for its detection and characterization in complex samples. We describe the development and application of the novel molecular tool for the specific enrichment of prokaryotic DNA. A fused protein comprising the DNA-binding subunit of the bacterial topoisomerase II, gyrase, was expressed, purified, and immobilized on magnetic particles. We demonstrated the specific affinity of the immobilized protein towards bacterial DNA and investigated its efficiency in the samples with high background of eukaryotic DNA. The reported approach allowed for the selective isolation and further detection of as few as 5 pg Staphylococcus aureus DNA from the sample with 4 × 106-fold surplus of human DNA. This method is a promising approach for the preparation of such type of samples, for example in molecular diagnostics of sepsis.

Keywords

Gyrase Bacterial DNA Target enrichment Bacteria detection 

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Copyright information

© Springer-Verlag Berlin Heidelberg 2014

Authors and Affiliations

  • Natalia Sandetskaya
    • 1
    Email author
  • Andreas Naumann
    • 1
  • Katharina Hennig
    • 2
  • Dirk Kuhlmeier
    • 1
  1. 1.Nanotechnology UnitFraunhofer Institute for Cell Therapy and Immunology IZILeipzigGermany
  2. 2.University of Applied Sciences JenaJenaGermany

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