A sequence alignment of the phlA gene from five 2,4-DAPG producing Pseudomonas strains revealed a very well conserved palindromic sequence GAAACN5GTTTC. EMSA verified binding of PsrA to the phlA promoter. Site-directed mutagenesis confirmed that binding was abolished when mutations were introduced into the putative PsrA binding site. phlA-lacZ transcriptional fusion in wild-type and psrA mutant strains confirmed that PsrA represses phlA expression. Western blot assay showed that levels of PhlA protein increased in the psrA mutant compared to the wild type strain.
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|