Genetic dissection of mycobacterial biofilms

Methods in Molecular Biology
Anil OjhaGraham F Hatfull

Abstract

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.

Citations

Apr 8, 2016·Expert Opinion on Pharmacotherapy·Concepcion Perez-JorgeJaime Esteban
Jun 20, 2018·Applied and Environmental Microbiology·Christopher EalandBavesh Kana
May 23, 2019·The Journal of Biological Chemistry·Prabhakar TiwariRamandeep Singh
Sep 11, 2019·Antimicrobial Agents and Chemotherapy·Jacob P RichardsAnil K Ojha
Feb 7, 2018·Frontiers in Microbiology·Jaime Esteban, Marta García-Coca
Jul 18, 2018·Indian Journal of Microbiology·Richa VirmaniYogendra Singh
Oct 28, 2019·The Journal of Biological Chemistry·Srijon Kaushik BanerjeeManikuntala Kundu
May 28, 2019·Antimicrobial Resistance and Infection Control·Divakar SharmaAsad U Khan
Apr 3, 2020·Journal of Microbiology and Biotechnology·Miguel Angel De la Cruz-VillegasJorge Bravo-Madrigal
Mar 4, 2021·MBio·J A JuddK M Derbyshire

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