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HutP-binding affinity of the site I and site II mutants of the truncated terminator RNA and the hut wild-type RNA, determined using a filter binding assay. (a) Base substitutions at site I and site II are indicated on the hut terminator RNA. (b) The equilibrium dissociation constant (Kd) for each mutant was determined. Binding reactions were performed with 10 nM of RNA and protein concentrations ranging from 250 to 2250 nM.
