EMSA performed with a series of the blh promoter fragments determined that BigR binds to a region containing an imperfect palindrome. DNase I footprinting confirmed that BigR binds to this palindrome. blh-EGFP reporter assays showed BigR acts as a repressor of the blh gene. Site-directed mutagenesis in conjunction with EMSA verified that mutations in the BigR binding site affected BigR binding. Multiple sequence alignment of the blh promoters from X. fastidiosa, A. tumefaciens, S. meliloti, M. loti, Chromobacterium violaceum, and Nitrosomonas europaea identified the BigR consensus sequence.
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|
|AATATATATTATTATATATTG||XF0768, XF0767, XF0766, XF0765, XF0764||