Construction of conditional knockdown mutants in mycobacteria

Abstract

By definition, essential genes are fundamental to bacterial growth, yet the functions of many such genes remain unknown. Essential genes furthermore are central to the activity of most antibacterial drugs and among the most attractive targets for the development of new therapeutics. This chapter describes how synthetic genetic switches that utilize transcriptional repression, controlled proteolysis, or both to silence gene activity can be applied to construct and characterize conditional knockdown (cKD) mutants for essential genes in Mycobacterium smegmatis and Mycobacterium tuberculosis.