First a DNase I footprint analysis showed that Fur binds specifically to a high-affinity site, then correct replacement of the wild-type sequence with the antibiotic resistance cassette was verified by means of PCR. The total RNA from the fur mutant strain was isolated, and transcription from the promoters under study was investigated by primer extension assays. Consensus sequence was also performed.
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|