Figure 2

Novel GlnR binding sites identified upstream of differentially expressed genes, with corresponding EMSA to confirm specific GlnR binding. EMSA were performed by incubating increasing amounts of His-GlnR recombinant protein with labelled DNA corresponding to the promoter regions of the genes downstream of the GlnR binding site. GlnR binding was visualised in IGV. The top track represents GlnR binding in nitrogen excess, the second track represents GlnR binding in nitrogen limiting conditions , and the third track represents input control DNA. Bar height is representative of fold change in gene expression in the WT compared to the ΔglnR mutant in nitrogen limitation. Levels of gene expression are indicated in the bottom track. Vertical lines through the peak indicate GlnR binding sites. (A). Peak 9, MSMEG2425 (amtB), (B). Peak 22, MSMEG2526, (C). Peak 17, MSMEG2184, (D). Peak 42, MSMEG5358, (E). Peak 21, MSMEG2522 and (F). Negative control, MSMEG3224.