Protocol

Mycobacteria Protocols

Volume 101 of the series Methods in Molecular Biology™ pp 235-243

The Application of Bacterial Luciferase as a Reporter Gene in Mycobacteria

  • Stephen GordonAffiliated withUnité de Génétique Moléculaire Bactérienne, Institut Pasteur
  • , Gavin ChungAffiliated withChemotherapy Department, Glaxo-Wellcome Medical Research Centre
  • , Peter AndrewAffiliated withDepartment of Microbiology and Immunology, University of Leicester

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Abstracts

Bioluminescence is the product of two distinct enzymes, firefly luciferase and bacterial luciferase. The application of the firefly enzyme in the study of mycobacteria has been described by other groups (1,2) (see Chapter 31). The bacterial-luciferase enzyme is a drmer of approx 80 kDa, consisting of an α- and β-subunit (3,4). It catalyzes the oxidation of long-chain fatty acids (>7 carbons) and reduced riboflavin, generating blue-green light (λ = 490 nm) in the process. The reaction can be written as follows: FMNH2 + RCHO + O2→ FMN + H2O + RCOOH + Light