A RADAR-Based Assay to Isolate Covalent DNA Complexes in Bacteria.

ABSTRACT: Quinolone antibacterials target the type II topoisomerases gyrase and topoisomerase IV and kill bacterial cells by converting these essential enzymes into cellular poisons. Although much is known regarding the interactions between these drugs and enzymes in purified systems, much less is known regarding their interactions in the cellular context due to the lack of a widely accessible assay that does not require expensive, specialized equipment. Thus, we developed an assay, based on the "rapid approach to DNA adduct recovery," or RADAR, assay that is used with cultured human cells, to measure cleavage complex levels induced by treating bacterial cultures with the quinolone ciprofloxacin. Many chemical and mechanical lysis conditions and DNA precipitation conditions were tested, and the meth

SUBMITTER: Aldred KJ 

PROVIDER: S-EPMC6466838 | biostudies-literature | 2019 Feb

REPOSITORIES: biostudies-literature