Research on the human pathogen Mycobacterium tuberculosis (Mtb) suffers from a dearth of genetic tools because systems from distantly related bacteria such as E. coli often do not work in Mtb.  We have created a new inducible mycobacterial expression system based on a synthetic riboswitch.  Unlike other systems, the riboswitch does not require repressor protein coexpression:  all the necessary machinery is on a single short stretch of DNA, making it easier to apply in a variety of contexts.  We have demonstrated in Mtb and related mycobacteria that this system can be used to (1) induce recombinant protein overexpression, (2) create a conditional gene knockout, and (3) control expression in Mtb in a macrophage infection model.    Finally, the small-molecule inducer is an approved drug, a feature that will facilitate its use in animal infection models.  This riboswitch-based system will thus be an important new tool for mycobacterial research.

Tools for manipulating gene expression in the human pathogen <i>Mycobacterium tuberculosis</i>

239th National Meeting (2010)

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Tools for manipulating gene expression in the human pathogen Mycobacterium tuberculosis