Fig. 5: Replication of genomic (−) and (+) strand under isolated reaction conditions. | Communications Biology

Fig. 5: Replication of genomic (−) and (+) strand under isolated reaction conditions.

From: In vitro characterisation of the MS2 RNA polymerase complex reveals host factors that modulate emesviral replicase activity

Fig. 5

Replication and fluorescence readout scheme for a [F30-Bro(+)]MS2(−) and b [F30-Bro(+)]MS2(+) synthesis. DFHBI-1T fluorescence of c [F30-Bro(+)]MS2(−) and d [F30-Bro(+)]MS2(+) synthesis after 3 h incubation in reaction buffer supplemented with MS2rep (1 µM) in the absence (red) of essential co-factors EF-Tu and EF-Ts (15 µM each), S1 (1.5 µM), or in the presence of all co-factors and IF1 (15 µM) (green). Positive controls (PC, blue) contained all co-factors except IF1. Negative controls (NC, grey) contained no MS2rep. Initial non-fluorescent template concentrations were 50 nM. e, f show the fluorescence time traces of both reactions in c, d recorded over five hours. Negative control (NC, red), positive control (PC, green), additional 15 µM IF1 (+IF1, orange). Error bars indicate standard deviations calculated from independent replicates from the same protein stocks.

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