Genetic dissection of mycobacterial biofilms

Anil K. Ojha, William R. Jacobs, Graham F. Hatfull

Research output: Contribution to journalArticlepeer-review

19 Scopus citations


Our understanding of the biological principles of mycobacterial tolerance to antibiotics is crucial for developing shorter anti-tuberculosis regimens. Various in vitro approaches have been developed to identify the conditions that promote mycobacterial persistence against antibiotics. In our laboratories, we have developed a detergent-free in vitro growth model, in which mycobacteria spontaneously grow at the air–medium interface as self-organized multicellular structures, called biofilms. Mycobacterial biofilms harbor a subpopulation of drug tolerant persisters at a greater frequency than their planktonic counterpart. Importantly, development of these structures is genetically programmed, and defective biofilms of isogenic mutants harbor fewer persisters. Thus, genetic analysis of mycobacterial biofilms in vitro could potentially be a powerful tool to unravel the biology of drug tolerance in mycobacteria. In this chapter we describe a method for screening biofilm-defective mutants of mycobacteria in a 96-well format, which readily yields a clonally pure mutant for further studies.

Original languageEnglish (US)
Pages (from-to)215-226
Number of pages12
JournalMethods in Molecular Biology
StatePublished - 2015
Externally publishedYes


  • Biofilm
  • Drug tolerance
  • Genetic mutants
  • Mycobacteria

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics


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