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Final compounds were cleaved from resign into 96-well plate and screened against S. aureus ATCC25293, E. coli ATCC25922 and E. coli MG1655. Hits identified from this screen were re-synthesized and their MIC's were determined against an expanded panel of organisms. E. coli MG1655 tolC is an efflux pump mutant.
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5 cfu/mL.
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36
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85031160573
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50 for negamycin 5.1 mg/kg.
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85031148006
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Inhibition of protein synthesis was measured with a cell-free transcription/translation coupled assay (E. coli S30 Extract System, Promega, Madison, WI, USA) using pGEMbgal as a DNA template. Compounds were pre-incubated with the reaction mixture for 10 min prior to the addition of template. After 1 h of incubation, ONPG was added and β-galactosidase activity was monitored at 420 nm.
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