Figure 8.
Effect of DnaN and/or PolCpol- on DnaE error-prone polymerase activity. Primer extension assays were carried out on RNA primed ssM13mp18 DNA omitting dGTP. Time course reactions at 30 and 60 min (indicated by rectangular triangles) were terminated by addition of loading dye and incubation at 95°C for 5 min. Primer extension products were analysed through 1.5% (w/v) alkaline agarose gels. (a) DnaE (360 nM) and (b) DnaE and PolCpol- at equimolar concentrations (360 nM) in the presence or absence of DnaN (360 nM), as indicated. Lanes labelled M and C show molecular weight markers and a control without polymerases, respectively. All assays were carried out in triplicates and the gels shown here are representative. Quantifications of nascent DNA synthesized (radioactivity measured in arbitrary units, AU) are shown in bar graphs underneath the gels.