Functional Analysis of the Role of Toxin–Antitoxin (TA) Loci in Bacterial Persistence

Part of the Methods in Molecular Biology book series (MIMB, volume 1333)


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.


Toxin–antitoxin Persistence Inducible expression E. coli Antibiotic 


  1. 1.
    Yamaguchi Y, Inouye M (2011) Regulation of growth and death in Escherichia coli by toxin-antitoxin systems. Nat Rev Microbiol 9:779–790CrossRefPubMedGoogle Scholar
  2. 2.
    Gerdes K, Maisonneuve E (2012) Bacterial persistence and toxin-antitoxin Loci. Annu Rev Microbiol 66:103–123CrossRefPubMedGoogle Scholar
  3. 3.
    Lewis K (2010) Persister cells. Annu Rev Microbiol 64:357–372CrossRefPubMedGoogle Scholar
  4. 4.
    Butt A, Higman VA, Williams C et al (2014) The HicA toxin from Burkholderia pseudomallei has a role in persister cell formation. Biochem J 459:333–334PubMedCentralCrossRefPubMedGoogle Scholar
  5. 5.
    Dörr T, Vulić M, Lewis K (2010) Ciprofloxacin causes persister formation by inducing the TisB toxin in Escherichia coli. PLoS Biol 8, e1000317PubMedCentralCrossRefPubMedGoogle Scholar
  6. 6.
    Korch S, Hill T (2006) Ectopic overexpression of wild-type and mutant hipA genes in Escherichia coli: effects on macromolecular synthesis and persister formation. J Bacteriol 188:3826–3836PubMedCentralCrossRefPubMedGoogle Scholar
  7. 7.
    Keren I, Kaldalu N, Spoering A, Lewis K (2004) Persister cells and tolerance to antimicrobials. FEMS Microbiol Lett 230:13–18CrossRefPubMedGoogle Scholar
  8. 8.
    Butt A, Muller C, Harmer N, Titball RW (2013) Identification of type II toxin-antitoxin modules in Burkholderia pseudomallei. FEMS Microbiol Lett 338:86–94CrossRefPubMedGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2016

Authors and Affiliations

  1. 1.Department of Infection and Immunity, Medical SchoolUniversity of SheffieldSheffieldUK
  2. 2.Biosciences, College of Life and Environmental SciencesUniversity of ExeterExeterUK

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