Revision as of 21:54, 21 October 2019

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.

Figure 1: The ratio of Fluorescence per OD between the temperatures of 30°C and 25°C. The horizontal dash line is used to compare the thermometers we made to that of the best performing kit thermometer.

Figure 2: The ratio of Fluorescence per OD between the temperatures of 37°C and 30°C. The horizontal dash line is used to compare the thermometers we made to that of the best performing kit thermometer.

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

Plant root growth was tested with different concentrations of arabinose because arabinose is the inducer for our enzyme circuits. Since arabinose is a sugar that the plant could metabolize, it was necessary to see how different concentrations of arabinose would affect plant growth without our enzyme circuits. Preliminary scans of the plates show sporadic root growth so we deemed the results unusable, and we are in the process of repeating these experiments.

Plant Root Growth with Auxin (IAA)

IAA (auxin) is a plant hormone that stimulates plant growth and root elongation. Since one of our enzyme circuits will upregulate the production of IAA, it was essential for us to have a control experiment of plant root growth with IAA. This will later be used to compare root growth when we grow plants with our enzyme circuits. Preliminary scans of the plates show sporadic root growth so we deemed the results unusable, and we are in the process of repeating these experiments.

Plant Root Growth with Trehalose

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Plant Root Growth with Arabinose, Auxin, and Trehalose at Maximum Concentration

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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.

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+     <p> <b>Figure 1</b>: The ratio of Fluorescence per OD between the temperatures of 30°C and 25°C. The horizontal dash line is used to compare the thermometers we made to that of the best performing kit thermometer.  </p>
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-     <p> Figure 2: </p>
+     <p> <b>Figure 2</b>: The ratio of Fluorescence per OD between the temperatures of 37°C and 30°C. The horizontal dash line is used to compare the thermometers we made to that of the best performing kit thermometer. </p>
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Difference between revisions of "Team:Rice/Results"