Team:Carnegie Mellon/Met-Results
From 2012.igem.org
Time course experiment with Spinach and FAP
The two figures below are plots of the experimental data from the time course experiments. The figures compare the fluorescence of the mutant promoters with the wild-type promoter, when either 3,5-difluoro-4-hydroxybenzylidene imidazolinone (DFHBI) or Malachite Green (MG) is added. As both DFHBI and MG are dyes that bind to RNA and protein specifically, there is a positive correlation between the fluorescence values and the amount of RNA and protein being produced in the cells.
Note that the fluorescence numbers plotted are divided by the OD600 readings to account for the different cell densities.
From the figures there are a few general observations that can be made. Firstly, for both the Spinach-DFHBI and FAP-MG plots, we see an increasing trend of the fluorescence values across all promoters. This makes sense, as we do expect the amount of RNA (Spinach) transcripted and protein (FAP) translated to increase with time.
Comparing the different promoters within the FAP-MG plot, the fluorescence level of Mutant I is increasing much more rapidly than the rest, indicating that this promoter significantly increases the translational rate of the cell.
Mutant II closely parallels the wild type fluorescence level, being only slightly lower in magnitude. Mutant III's fluorescence level increases very rapidly at first, but seems to be leveling off after an hour. This may indicate that the cell's peak translational rate has already been achieved.
Dosage curves of Spinach and FAP with their respective dyes