Development and Application of Synthetic Riboswitches as Tools to Study Bacterial Pathogenesis

In the laboratory, we require tools that allow conditional regulation of gene expression to facilitate the study of genes with unknown or little understood function. The ideal tool would provide translation of external stimuli to an internal alteration in gene expression for any gene to be studied. Furthermore, these tools should be transferrable between different species of bacteria to allow manipulation of expression in less understood prokaryotic species. Nature has the advantage of billions of years of evolution to determine efficient mechanisms for gene regulation. We have looked to nature for inspiration and adapted natural systems to serve alternative purposes in the laboratory. In this thesis we describe the development of synthetic riboswitches as orthogonal tools to conditionally express bacterial genes of interest. We will explore the concept of isolating functional riboswitches from a genetically tractable species of bacterial and transporting these tools with limited alteration to less genetically tractable species. Each chapter focuses on a different bacterial species with relevance to different areas of microbiology and biotechnology and presents what we have learned regarding the portability of synthetic riboswitches.