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. 2016 Apr 14;8(4):100. doi: 10.3390/v8040100

Figure 5.

Figure 5

Kinetics of template and substrate utilisation by calicivirus RdRps. Purified recombinant RdRps (40 ng of RHDV and RCV RdRps and 200 ng of NoV RdRp) were used to generate dsRNA with poly(C) RNA as template and rGTP as substrate. (a) The kinetics of template utilisation was examined by titrating poly(C) from 0 to 40 ng/μL in the presence of 0.5 mM rGTP; (b,c) The kinetics of nucleotide incorporation was examined by titrating rGTP from 0 to 0.5 mM and 0 to 2.0 mM in the presence of 20 ng/μL of poly(C). Following a 15-min incubation at 30 °C, reactions were stopped with EDTA at final concentration 5 mM and dsRNA was quantified using the PicoGreen reagent. RHDV RdRp activity is shown in red as triangles, RCV RdRp activity is shown in blue as squares and NoV RdRp activity is shown in black as circles. The results from a representative experiment are shown with average values and standard deviations from triplicate reactions for each measurement point.