Related Experiment Video
Updated: Jan 5, 2026

Using Mycobacterium smegmatis as a Bioindicator for Zinc-Limited Growth Conditions in Mycobacteria
Published on: September 20, 2024
The use of temperature-sensitive plasmids in mycobacteria
Damien Portevin1, Wladimir Malaga, Christophe Guilhot
1Mycobacterial Division Research, National Institute for Medical Research, London, UK.
Abstract:
The construction of allelic exchange mutants is one of the major strategies to decipher the function of a defined gene. In this chapter, protocols are described to perform allelic exchange in the mycobacterial species Mycobacterium smegmatis and Mycobacterium tuberculosis using thermosensitive counterselectable vectors. The antibiotic resistance cassette used makes it possible to efficiently rescue the marker, leaving an unmarked mutation on the chromosome. In addition, a method is provided to construct conditional M. smegmatis mutants and to test whether or not a gene is essential.
Related Concept Videos
Factors Influencing Microbial Growth: Temperature
Hyperthermophilic Bacteria
Antibiotic Selection

