Figure 8
Blocking the synthesis of CSPA:GFP11 uncouples the complementation system. (A) In vitro translation of cspA:gfp11 mRNA (60 pmol) was performed in the presence of increasing concentrations of Kanamycin at 37 °C for 30 min. At the end of incubation, 800 pmol of MBP:GFP1-10fast were added to each reaction tube, which were further incubated at 25 °C for 2 h before measuring fluorescence. (B) Fluorescence signal measured when 800 pmol of MBP:GFP1-10fast were added to: (i) sample translated in the absence of antibiotic (red), (ii) sample translated in the absence of antibiotic with simultaneous addition of 50 µM Kanamycin and MBP:GFP1-10fast (blue), (iii) sample translated in the presence of 50 µM Kanamycin (black). The interaction between the two GFP fragments was conducted at 25 °C for 2 h. The error bars represent the standard error calculated from triplicates. In all experiments, the fluorescence signal displayed by control tubes that did not contain mRNA was subtracted.
