Arizona_State
Improve
The improvement of F2620 receiver: This year's team conducted multiple experiments to re-characterize a an improved part of F2620. The improved part itself can also be viewed here on the iGEM registry page.
There are 3 types of experiments used to characterize this improved receiver: F2620 Induced by Synthetic AHLs, F2620 Induced by Sender Supernatants, and F2620 induced by Sender cells on Agar.
The first experiment, F2620 Induced by Synthetic AHLs, included F2620 receiver cells being induced by synthetic AHL chemicals under different concentrations. The maximum GFP expression was analyzed to understand the effect AHL concentration had on the receiver expressing GFP.
The following experiment, F2620 Induced by Sender Supernatants, used F2620 receiver cells being induced by a variety of combinations of senders in supernatant form. The maximum GFP expression was analyzed to understand the effect different percentages of sender supernatant had on the receiver expressing GFP.
The final experiment included F2620 receiver cells that were spread on agar plates and induced by spread sender cells. Images are then taken at different time intervals of the agar plate to analyze the induction rates and diffusion of F2620 induction.
F2620 Induced by Synthetic AHLs
The graphs below depict the GFP production from the F2620 receiver to be higher at a higher concentration of the synthetic AHL of Las. The second graph, depicts the same nature however at a higher concentration of the synthetic AHL of Lux. It is interesting to note that the Las sender, at a lower concentration, promotes a higher GFP expression than the Lux. Our group hypothesized that it would be opposite considering the LuxI sender comes from the same system as F2620. These results show differently.
F2620 Induced by Sender Supernatants
When testing the LuxR the AubI showed a higher expression when mixed with 10% of a second sender (even when that sender was a negative control sender). The 40% AubI mixed with 10% negative sender and the 40% AubI mixed with 10% RhlI both expressed higher than the 50% AubI by itself. This result was confirmed in another test where 40% Aub mixed with 10% EsaI and 40% AubI mixed with 10% CerI both expressed higher than the 50% AubI alone. (See graphs below).
F2620 induced by Sender cells on Agar
