title: Functional Analysis of the Role of Toxin-Antitoxin (TA) Loci in Bacterial Persistence.
abstract: We have developed a method to analyze the functionality of putative TA loci by expressing them in Escherichia coli. Here, we describe the procedure for cloning recombinant TA genes into inducible plasmids and expressing these in E. coli. Following expression, toxicity, resuscitation of growth, and changes in persister cell formation are assayed. This can confirm whether predicted TA loci are active in E. coli and whether expression can affect persister cell formation.
authors.affiliation: Department of Infection and Immunity, Medical School, University of Sheffield, Beech Hill Road, Sheffield, S10 2RX, UK.
authors.forename: Aaron T
authors.initials: AT
authors.lastname: Butt
authors.affiliation: Biosciences, College of Life and Environmental Sciences, University of Exeter, Geoffrey Pope Building, Stocker Road, Exeter, EX4 4QD, UK. r.w.titball@exeter.ac.uk.
authors.forename: Richard W
authors.initials: RW
authors.lastname: Titball
mesh_terms: D000992:Antitoxins / Q000096:biosynthesis / Q000235:genetics*; D001427:Bacterial Toxins / Q000096:biosynthesis / Q000235:genetics*; D003001:Cloning, Molecular / Q000379:methods*; D004926:Escherichia coli / Q000187:drug effects / Q000235:genetics*; D015964:Gene Expression Regulation, Bacterial / Q000187:drug effects
chemical_list: D000992:Antitoxins; D001427:Bacterial Toxins
keywords: Antibiotic; E. coli; Inducible expression; Persistence; Toxin–antitoxin