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. Author manuscript; available in PMC: 2012 Jan 21.
Published in final edited form as: J Mol Biol. 2010 Oct 28;405(3):666–678. doi: 10.1016/j.jmb.2010.10.020

Fig. 3. Elongation substrate with 2AP in the nontemplate strand monitors the kinetics of CMP incorporation.

Fig. 3

(A) The G/2AP-NTn elongation substrate contains a 2AP in the NT at the n-position and correctly base pairs with CTP. (B) A representative time trace of 2AP fluorescence change at 40 μM CTP (200nM G/2AP-NTn) and 400 nM T7 RNAP). The experimental data fit well to a single exponential equation (Eq.1) (solid line) with kobs= 136 ± 24 s-1. (D) The kobs increased with increasing [CTP] and fit to a hyperbola (Eq. 2) with a maximum rate constant kpol = 209 ± 27 s-1 and Kd = 28 ± 13 μM. Error bars indicate the high and low values from duplicate measurements.