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Benzoic Acid-Inducible Gene Expression in Mycobacteria.


ABSTRACT: 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.

SUBMITTER: Dragset MS 

PROVIDER: S-EPMC4562662 | biostudies-literature | 2015

REPOSITORIES: biostudies-literature

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Benzoic Acid-Inducible Gene Expression in Mycobacteria.

Dragset Marte S MS   Barczak Amy K AK   Kannan Nisha N   Mærk Mali M   Flo Trude H TH   Valla Svein S   Rubin Eric J EJ   Steigedal Magnus M  

PloS one 20150908 9


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 no  ...[more]

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