An array of target-specific screening strains for antibacterial discovery.
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.
DeVito
Joseph A
JA
Department of Antimicrobial Research, Bristol-Myers Squibb Company, Wilmington, DE 19880, USA.
Mills
Jonathan A
JA
Liu
Veronica G
VG
Agarwal
Anjana
A
Sizemore
Christine F
CF
Yao
Zhongjie
Z
Stoughton
Daniel M
DM
Cappiello
Maria Grazia
MG
Barbosa
Maria D F S
MD
Foster
Lorie A
LA
Pompliano
David L
DL
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
D000900:Anti-Bacterial Agents; D019883:Alkyl and Aryl Transferases; C022566:UDP-N-acetylglucosamine 1-carboxyvinyltransferase