Primer extension was used to determine the gene start-site and show that there was promoter activity between +50 and -150. Two candidate sites were identified using a cited consensus sequence, and were confirmed via B-gal assay to upregulate flhDC. Binding was then confirmed with site directed mutagenesis and a series of EMSA's. The EMSAs showed significantly more binding at site 2 than site 1.
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|