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Suppressor mutant exhibits comparable DNA-binding affinity. EMSA analyses for the DNA-binding affinity of WT KpnI and mutants. Different concentrations of WT or KpnI mutants (0–256 nM) were incubated with 1 nM of end-labeled canonical (-GGTACC) oligonucleotide in binding buffer [20 mM Tris–HCl (pH 7.4), 25 mM NaCl and 5 mM 2-mercaptoethanol] on ice for 15 min before loading onto a 8% polyacrylamide gel. The intensity of the shifted DNA band is expressed as percentage bound.
