FIGURE 9.
a, fluorescence titrations of poly(ϵA) with the DENV polymerase (λex = 325 nm, λem = 410 nm) in buffer T5 (pH 7. 6, 10 °C), containing 1 mm MgCl2, in the absence (■) and presence (□) of the dsRNA 30-mer (see “Experimental Procedures”). The concentration of poly(ϵA) is 9.41 × 10−6 m (nucleotide). The concentration of the dsRNA 30-mer is 1.44 × 10−5 m (oligomer). The solid lines are nonlinear least squares fits of the binding titration curves, using Equations 6, 7, 13, and 14, with K = 2.5 × 106 m−1, ω = 14, and ΔFmax = 1.19 for poly(ϵA) and KS ≤ 1 × 105 m−1 for the dsRNA oligomer. b, fluorescence titrations of poly(ϵA) with the DENV polymerase (λex = 325 nm, λem = 410 nm) in buffer T5 (pH 7.6, 10 °C), containing 1 mm MgCl2, in the absence (■) and presence (□) of the dsDNA 30-mer (see “Experimental Procedures”). The concentration of poly(ϵA) is 9.41 × 10−6 m (nucleotide). The concentration of the dsDNA 30-mer is 1.0 × 10−5 m (oligomer). The solid lines are nonlinear least squares fits of the binding titration curves, using Equations 8, 9, 15, and 16, with K = 2.5 × 106 m−1, ω = 14, and ΔFmax = 1.19 for poly(ϵA) and KS = 8.5 × 105 m−1 for the dsDNA oligomer.