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DIG-labeled pyhaO was incubated with increasing concentrations of YhaJ that corresponded to a shift in free-DNA indicating a YhaJ-DNA complex. Specificity of the binding reaction was tested by the addition of a 100-fold excess (+) of unlabeled pyhaO probe to the binding reaction to outcompete binding of the DIG-labeled probe to YhaJ. These reaction conditions were carried out using a fragment of the kan gene as a negative control. Additionally, the unlabeled kan probe in 100-fold excess was used as a non-specific competitor for YhaJ binding to the DIG-labeled yhaO probe (pyhaO vs kan). (B) Activity of the yhaO promoter in the ΔyhaJ mutant background. A plasmid containing a GFP-yhaO promoter fusion was transformed into TUV93-0 and ΔyhaJ to monitor transcription of yhaO in RFU during growth in MEM-HEPES. 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