Figure 5.

Demonstration of ribosome modification in operon-based iSAT system. The point mutation A2058U of the 23S rRNA gene was made to pT7rrnB (WT) to confer resistance to the antibiotic clindamycin, and the resulting plasmid, pT7rrnB-CR (CR), was used in iSAT reactions. Clindamycin was added to the appropriate iSAT reactions at a final concentration of 50 ng/μl prior to incubation, and activity was assessed through luciferase production at 37°C for 4 h. WT iSAT reactions showed an average luciferase concentration of less than 0.25 nM in the presence of clindamycin. Values show average luciferase concentrations above background with error bars representing s.d. for three independent reactions.