title: An array of target-specific screening strains for antibacterial discovery.
abstract: As the global threat of drug- and antibiotic-resistant bacteria continues to rise, new strategies are required to advance the drug discovery process. This work describes the construction of an array of Escherichia coli strains for use in whole-cell screens to identify new antimicrobial compounds. We used the recombination systems from bacteriophages lambda and P1 to engineer each strain in the array for low-level expression of a single, essential gene product, thus making each strain hypersusceptible to specific inhibitors of that gene target. Screening of nine strains from the array in parallel against a large chemical library permitted identification of new inhibitors of bacterial growth. As an example of the target specificity of the approach, compounds identified in the whole-cell screen for MurA inhibitors were also found to block the biochemical function of the target when tested in vitro.
authors: Department of Antimicrobial Research, Bristol-Myers Squibb Company, Wilmington, DE 19880, USA.
authors: Joseph A
authors: JA
authors: DeVito
authors: Jonathan A
authors: JA
authors: Mills
authors: Veronica G
authors: VG
authors: Liu
authors: Anjana
authors: A
authors: Agarwal
authors: Christine F
authors: CF
authors: Sizemore
authors: Zhongjie
authors: Z
authors: Yao
authors: Daniel M
authors: DM
authors: Stoughton
authors: Maria Grazia
authors: MG
authors: Cappiello
authors: Maria D F S
authors: MD
authors: Barbosa
authors: Lorie A
authors: LA
authors: Foster
authors: David L
authors: DL
authors: Pompliano
mesh_terms: D019883:Alkyl and Aryl Transferases / Q000378:metabolism; D000483:Alleles; D000900:Anti-Bacterial Agents / Q000494:pharmacology*; D004353:Drug Evaluation, Preclinical / Q000379:methods*; D004926:Escherichia coli / Q000378:metabolism; D020128:Inhibitory Concentration 50; D007700:Kinetics; D008957:Models, Genetic; D020411:Oligonucleotide Array Sequence Analysis; D010957:Plasmids / Q000378:metabolism; D011487:Protein Conformation; D011995:Recombination, Genetic; D013997:Time Factors
chemical_list: D000900:Anti-Bacterial Agents; D019883:Alkyl and Aryl Transferases; C022566:UDP-N-acetylglucosamine 1-carboxyvinyltransferase