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RNA binding analysis of recombinant PfPuf1 RNA-binding domain. (A) A gel retardation assay using ∼30 fmol of 32P-labeled NRE RNA and various concentrations (0–240 nM) of recombinant PfPuf1. The Kd was estimated to be <20 nM. (B) A gel retardation assay using ∼30 fmol of 32P-labeled NRE RNA and 60 nM recombinant PfPuf1 in the presence of a 10–100× excess of cold competitor RNAs. NRE and three mutants (Box A, Box B and both Boxes A and B) were included as competitors.
