title: Experimental Evolution of Escherichia coli Persister Levels Using Cyclic Antibiotic Treatments.
abstract: Persister cells are difficult to study owing to their transient nature and their usually small number in bacterial populations. In the past, numerous attempts have been made to elucidate persistence mechanisms. However, because of the challenges involved in studying persisters and the clear redundancy in mechanisms underlying their generation, our knowledge of molecular pathways to persistence remains incomplete. Here, we describe how to use experimental evolution with cyclic antibiotic treatments to generate mutants with an increased persister level in stationary phase, ranging from the initial ancestral level up to 100 %. This method will help to unravel molecular pathways to persistence, and opens up a myriad of new possibilities in persister research, such as the convenient study of nearly pure persister cultures and the possibility to investigate the role of time and environmental aspects in the evolution of persistence.
authors.affiliation: Centre of Microbial and Plant Genetics (CMPG), KU Leuven - University of Leuven, Leuven, 3001, Belgium.
authors.forename: Bram
authors.initials: B
authors.lastname: Van den Bergh
authors.affiliation: Centre of Microbial and Plant Genetics (CMPG), KU Leuven - University of Leuven, Leuven, 3001, Belgium.
authors.forename: Joran E
authors.initials: JE
authors.lastname: Michiels
authors.affiliation: Centre of Microbial and Plant Genetics (CMPG), KU Leuven - University of Leuven, Leuven, 3001, Belgium. jan.michiels@biw.kuleuven.be.
authors.forename: Jan
authors.initials: J
authors.lastname: Michiels
mesh_terms: D000222:Adaptation, Physiological / Q000235:genetics; D000617:Aminoglycosides / Q000235:genetics / Q000378:metabolism; D000900:Anti-Bacterial Agents / Q000494:pharmacology*; D019020:Directed Molecular Evolution / Q000379:methods*; D004926:Escherichia coli / Q000187:drug effects* / Q000235:genetics* / Q000254:growth & development; D053858:Metabolic Networks and Pathways / Q000187:drug effects; D008826:Microbial Sensitivity Tests / Q000379:methods; D009154:Mutation
chemical_list: D000617:Aminoglycosides; D000900:Anti-Bacterial Agents
keywords: Adaptation; Aminoglycosides; Bet–hedging; Cyclic treatments; Escherichia coli; Experimental evolution; Persister mutants