Team:Goettingen/Team/Reporter

<p>The activity of transcriptional regulators can be easily detected by monitoring the activity of a promoter that is fused to a reporter gene such as the green fluorescent protein (GFP). Recently, in <i>Mycobacterium smegmatis</i>, the transcriptional repressor DarR was identified (Zhang <i>et al.</i> 2013). DarR is capable of binding to a specific DNA sequence upon association with c-di-AMP. This led us to the idea of developing a powerful screening system to facilitate the identification of potential drugs interfering with c-di-AMP biofunction. While many Gram-positive bacteria (also pathogenic bacteria that cause severe human diseases) require c-di-AMP for growth, c-di-AMP is not synthesized by the Gram-negative bacterium <i>Escherichia coli</i>. Thus, c-di-AMP is not needed for growth of <i>E. coli</i>. Therefore, we intend to build a reporter system that is composed of existing and of novel biobricks that will be developed during the project (Figure 1). As <i>E. coli</i> does not require c-di-AMP for growth the screening system can be used to sort out those molecules from a drug library that interfere with general cellular processes (i.e. replication). By contrast, the system is suitable to identify compounds that specifically interfere with the signaling function of c-di-AMP. </p>