In Vitro Assays to Identify Antibiotics Targeting DNA Metabolism

Methods Mol Biol. 2017:1520:175-200. doi: 10.1007/978-1-4939-6634-9_11.


DNA metabolism embodies a number of biochemical pathways, which include targets of clinically used antibiotics as well as those that are only being explored as potential targets for inhibitory compounds. We give an overview of representative cell-based and enzymatic assays suitable for high-throughput-driven search for novel DNA metabolism inhibitors of established and novel DNA metabolism targets in bacteria. The protocol for a colorimetric coupled primase-inorganic pyrophosphatase assay developed by our group is described in detail.

Keywords: DNA recombination; DNA replication; High-throughput assay; Inhibitor discovery; Resistance.

MeSH terms

  • Anti-Bacterial Agents / pharmacology*
  • Biological Assay / methods*
  • DNA, Bacterial / metabolism*
  • High-Throughput Screening Assays
  • Indicators and Reagents
  • Inorganic Pyrophosphatase / metabolism
  • Mycobacterium tuberculosis / enzymology
  • Rosaniline Dyes / chemistry


  • Anti-Bacterial Agents
  • DNA, Bacterial
  • Indicators and Reagents
  • Rosaniline Dyes
  • malachite green
  • Inorganic Pyrophosphatase