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A Novel Fluorescence Based Screen for Inhibitors of the Initiation of DNA Replication in Bacteria.
Current Drug Discovery Technologies 2018 April 23
BACKGROUND: One of many strategies to overcome antibiotic resistance is the discovery of compounds targeting cellular processes, which have not yet been exploited.
METHODS AND MATERIALS: Using various genetic tools, we constructed a novel high throughput, cell based, fluorescence screen for inhibitors of chromosome replication initiation in bacteria.
RESULTS: The screen was validated by expression of an intra-cellular cyclic peptide interfering with the initiator protein DnaA and by over-expression of the negative initiation regulator SeqA. We also demonstrated that neither tetracycline nor ciprofloxacin triggers a false positive result. Finally, 400 extracts isolated mainly from filamentous actinomycetes were subjected to the screen.
CONCLUSION: We concluded that the presented screen is applicable for identifying putative inhibitors of DNA replication initiation in a high throughput setup.
METHODS AND MATERIALS: Using various genetic tools, we constructed a novel high throughput, cell based, fluorescence screen for inhibitors of chromosome replication initiation in bacteria.
RESULTS: The screen was validated by expression of an intra-cellular cyclic peptide interfering with the initiator protein DnaA and by over-expression of the negative initiation regulator SeqA. We also demonstrated that neither tetracycline nor ciprofloxacin triggers a false positive result. Finally, 400 extracts isolated mainly from filamentous actinomycetes were subjected to the screen.
CONCLUSION: We concluded that the presented screen is applicable for identifying putative inhibitors of DNA replication initiation in a high throughput setup.
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