Virtual Footprint program identified a putative CRP binding site in the promoter regions of luxR and ainS genes. luxR-gfp and ainS-gfp reporter assays showed that a 15 and 10 fold decrease in fluorescence in the crp mutant compared to the wild type strain. Quantitative real-time PCR showed that luxR and ainS transcript level was decreased by 250- and 7- fold, repctively, in the crp mutant strain compared to the wild type V. fischeri ES114. Fluorescence polarization DNA binding assays confirmed that CRP binds upstream of luxR and ainS.
Regulated genes for each binding site are displayed below. Gene regulation diagrams show binding sites, positively-regulated genes, negatively-regulated genes, both positively and negatively regulated genes, genes with unspecified type of regulation. For each indvidual site, experimental techniques used to determine the site are also given.
|Site sequence||Regulated genes||Gene diagram||Experimental techniques||TF function||TF type|