RNA Thermometer Results
To test the existing kit thermometers and our computationally designed thermometers in vivo, we made an RNA thermometer testing circuit, transformed it into E. coli, and tested the fluorescent output at different temperatures. These graphs show the relative fluorescent intensity of each thermometer between either 25 °C and 30 °C or 25 °C and 37 °C. Only thermometers that performed better than the positive control were included. "K" stands for the kit thermometers, and "NC" stands for our computationally designed NoChill series of thermometers. For additional thermometer testing data, please reference the contribution page.
As the first graph shows, most thermometers had about a two-fold change in expression between 25 °C and 30 °C. Additionally, most did not perform better than the existing kit thermometer. Since a 5 °C change in temperature is not that much, we also tested the difference between 25 °C and 37 °C, and the second graph shows that most of the thermometers in that experiment had around a four to six–fold expression change.
Plant Growth Results
Determining Plant Media Conditions
The lab that helped us with our plant experiments grewArabidopsis thaliana in PNS media. However, we wanted to make sure that our Pseudomonas putida would also be able to grow in the media, because it cannot use sucrose as the sole carbon source. We tested the effect of sodium, sodium citrate, glucose, and glucose+sodium on Pseudomonas growth. The bacteria grew in all conditions except for just sodium, so for ease, we decided to add glucose to our plant medium.
We needed to make sure that adding glucose would not adversely affect plant growth. To do this we tested the effect of two different glucose concentrations on plant and bacteria growth and found that the plants grew noticebly more at 0.01% glucose.
Plant Root Growth with Arabinose
10 μM and 50 μM
Plant Root Growth with Auxin (IAA)
10 μM
Plant Root Growth with Trehalose
Plant Root Growth with Arabinose, Auxin, and Trehalose at Maximum Concentration
Arabinose 50 mM, Auxin 10 uM, Trehalose 10 mM
Flood Inoculation of A. thaliana with P. putida in 0.6% PN and glucose
0.1% and 0.01% glucose pics here
Testing A. thaliana with induced and uninduced P. putida containing enzyme constructs
pIG104, non-induced and induced (insert pic below)
pIG105, non-induced and induced(insert pic below)
pIG106, non-induced and induced(insert pic below)
pIG104, pIG105, pIG106, non-induced and induced(insert pic below)
No bacteria(insert pic below)
Conclusion
This is the conclusion of our results. The data above demonstrate the validity of our project idea.