Antibiotic Combinations That Enable One-Step, Targeted Mutagenesis of Chromosomal Genes.

ABSTRACT: Targeted modification of bacterial chromosomes is necessary to understand new drug targets, investigate virulence factors, elucidate cell physiology, and validate results of -omics-based approaches. For some bacteria, reverse genetics remains a major bottleneck to progress in research. Here, we describe a compound-centric strategy that combines new negative selection markers with known positive selection markers to achieve simple, efficient one-step genome engineering of bacterial chromosomes. The method was inspired by the observation that certain nonessential metabolic pathways contain essential late steps, suggesting that antibiotics targeting a late step can be used to select for the absence of genes that control flux into the pathway. Guided by this hypothesis, we have identified anti

SUBMITTER: Lee W 

PROVIDER: S-EPMC5993608 | biostudies-literature | 2018 Jun

REPOSITORIES: biostudies-literature