title: In Vitro Assays to Identify Antibiotics Targeting DNA Metabolism.
abstract: 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.
authors.affiliation: Department of Pharmaceutical Sciences, University of Kentucky, 789 S. Limestone Street, Lexington, KY, 40536, USA.
authors.forename: Allan H
authors.initials: AH
authors.lastname: Pang
authors.affiliation: Department of Pharmaceutical Sciences, University of Kentucky, 789 S. Limestone Street, Lexington, KY, 40536, USA.
authors.forename: Sylvie
authors.initials: S
authors.lastname: Garneau-Tsodikova
authors.affiliation: Department of Pharmaceutical Sciences, University of Kentucky, 789 S. Limestone Street, Lexington, KY, 40536, USA. ovts222@uky.edu.
authors.forename: Oleg V
authors.initials: OV
authors.lastname: Tsodikov
mesh_terms: D000900:Anti-Bacterial Agents / Q000494:pharmacology*; D001681:Biological Assay / Q000379:methods*; D004269:DNA, Bacterial / Q000378:metabolism*; D057166:High-Throughput Screening Assays; D007202:Indicators and Reagents; D043564:Inorganic Pyrophosphatase / Q000378:metabolism; D009169:Mycobacterium tuberculosis / Q000201:enzymology; D012394:Rosaniline Dyes / Q000737:chemistry
chemical_list: D000900:Anti-Bacterial Agents; D004269:DNA, Bacterial; D007202:Indicators and Reagents; D012394:Rosaniline Dyes; C005095:malachite green; D043564:Inorganic Pyrophosphatase
keywords: DNA recombination; DNA replication; High-throughput assay; Inhibitor discovery; Resistance