Benzoic Acid-Inducible Gene Expression in Mycobacteria.
Conditional expression is a powerful tool to investigate the role of bacterial genes. Here, we adapt the Pseudomonas putida-derived positively regulated XylS/Pm expression system to control inducible gene expression in Mycobacterium smegmatis and Mycobacterium tuberculosis, the causative agent of human tuberculosis. By making simple changes to a Gram-negative broad-host-range XylS/Pm-regulated gene expression vector, we prove that it is possible to adapt this well-studied expression system to non-Gram-negative species. With the benzoic acid-derived inducer m-toluate, we achieve a robust, time- and dose-dependent reversible induction of Pm-mediated expression in mycobacteria, with low background expression levels. XylS/Pm is thus an important addition to existing mycobacterial expression tools, especially when low basal expression is of particular importance.
Department of Immunology and Infectious Diseases, Harvard School of Public Health, Boston, Massachusetts, United States of America; Centre of Molecular Inflammation Research, Department of Cancer Research and Molecular Medicine, Norwegian University of Science and Technology, Trondheim, Norway; Department of Biotechnology, Norwegian University of Science and Technology, Trondheim, Norway.
Marte S
MS
Dragset
Massachusetts General Hospital, Department of Medicine, Boston, Massachusetts, United States of America.
Amy K
AK
Barczak
Centre of Molecular Inflammation Research, Department of Cancer Research and Molecular Medicine, Norwegian University of Science and Technology, Trondheim, Norway.
Nisha
N
Kannan
Centre of Molecular Inflammation Research, Department of Cancer Research and Molecular Medicine, Norwegian University of Science and Technology, Trondheim, Norway.
Mali
M
Mærk
Centre of Molecular Inflammation Research, Department of Cancer Research and Molecular Medicine, Norwegian University of Science and Technology, Trondheim, Norway.
Trude H
TH
Flo
Department of Biotechnology, Norwegian University of Science and Technology, Trondheim, Norway.
Svein
S
Valla
Department of Immunology and Infectious Diseases, Harvard School of Public Health, Boston, Massachusetts, United States of America.
Eric J
EJ
Rubin
Department of Immunology and Infectious Diseases, Harvard School of Public Health, Boston, Massachusetts, United States of America; Centre of Molecular Inflammation Research, Department of Cancer Research and Molecular Medicine, Norwegian University of Science and Technology, Trondheim, Norway; Central Norway Regional Health Authority, Stjørdal, Norway.
Magnus
M
Steigedal
D001426:Bacterial Proteins / Q000096:biosynthesis / Q000235:genetics; D019817:Benzoic Acid / Q000494:pharmacology; D015964:Gene Expression Regulation, Bacterial; D005822:Genetic Vectors; D006801:Humans; D020102:Mycobacterium smegmatis / Q000235:genetics* / Q000472:pathogenicity; D009169:Mycobacterium tuberculosis / Q000235:genetics* / Q000472:pathogenicity; D011401:Promoter Regions, Genetic; D016958:Pseudomonas putida / Q000235:genetics*; D015534:Trans-Activators / Q000096:biosynthesis / Q000235:genetics; D014376:Tuberculosis / Q000235:genetics* / Q000382:microbiology
D001426:Bacterial Proteins; D015534:Trans-Activators; C078720:XylS protein, Pseudomonas putida; D019817:Benzoic Acid