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Performance Data

Heat Shock on Temperature Sensor/Actuator

. It was found that E.coli transformed with the TSA expressed Comet at different amounts for each temperature of heat shock relative to baseline (without heat shock). Comet expression at 40°C, 43°C, 46°C and at 49°C were similar over the first 3 hours. After the third hour, while Comet expression in the experimental group that were heat shocked at 40°C, 43°C, and 49°C steadily decreased, Comet expression in the experimental group that was heat shocked at 46°C increased. The level of Comet expression for the experimental group of cells that were heat shocked at 46°C approached the levels of Comet expression achieved by the E.coli cells in the first experiment that were heat shocked at 45°C. This suggested that there is a small temperature window at which E.coli transformed with the TSA upregulates Comet expression in response to heat shock relative to its baseline expression.

Heat Shock on Control

. In our positive control group, E.coli did not change their Comet expression compared to baseline at 43°C, 45°C, and at 46°C, though there was a slight increase in the group of cells that were heat shocked at 40°C and 49°C. This appeared to indicate that weak and severe heat shock slightly upregulated Comet expression in E.coli transformed with the p82 plasmid while temperatures normally considered as heat shock temperatures for E.coli induced little increase in Comet expression relative to its baseline. Nonetheless the findings of the positive control and experimental group show that Comet expression increases more relative to its baseline in response to heat shock in E.coli transformed with the TSA than those transformed with the p82 plasmid.

"Cold Shock on Temperature Sensor/Actuator"

After confirming transformation was successful of our positive and experimental plasmid, we examined the meffBlue expression of our experimental and positive control group overtime after being cold shocked at 10°C. It was found that in the absence of cold shock, the TSA plasmid has mild absorbance at 592 nm. After taking this into consideration, we observed only a modest increase in absorbance at 592 nm over a five-hour time frame following cold shock in the experimental group. Oddly, the positive control cells had a higher absorbance at 592 nm than the experimental group and increased for two and a half hours before decreasing. Comparing the responses of the positive control to our experimental group allowed us to determine that perhaps ten degrees was not the appropriate cold shock temperature to observe adequate changes in meffBlue expression compared to baseline.

In the second experiment, we analyzed the meffBlue expression of our experimental and positive control group at 4°C, 12°C, and 7°C. It was found that E.coli transformed with the TSA expressed meffBlue at different amounts for each temperature of cold shock relative to baseline (without cold shock). MeffBlue expression started similar to baseline after cold shock, regardless of cold shock temperature. Afterwards, meffBlue expression continued to increase across all cold shock temperatures, including baseline, until two and a half hours post cold shock. MeffBlue expression decreased thereafter and five hours post cold shock, meffBlue expression across all cold shock temperatures, including baseline were of a similar level. After comparing the two plates, it appears that the TSA plasmid works and meffBlue is expressed in response to cold shock, however, quantifying the cold shock response as absorbance at 592 nm was not ideal.

"Response to Multiple Heat Shocks"

In the third experiment we analyzed whether Comet expression would change if E.coli transformed with the TSA plasmid was exposed to repeated heat shocks at 45°C. After analyzing the data, it appears that multiple heat shocks slightly increases the Comet expression levels in the experimental group . However, after receiving more than 6 equally spaced heat shocks at 45°C over a three hour time frame, there is a slight decrease in Comet expression in the experimental group. It seems that there is a limit of heat shocks the E.coli transformed with the TSA plasmid can receive before Comet expression ceases to increase. This is supported by the data collected from the positive control because, as expected, there is no correlation between the number of heat shocks and Comet expression. It appears that repeated heat shock for the experimental and positive control group did not allow for the E.coli to recover sufficiently to change Comet expression in response to heat shock.

"Response to Multiple Cold Shocks"

In the fourth experiment we analyzed whether meffBlue expression would change if E.coli transformed with the TSA plasmid was exposed to repeated cold shocks at 10°C. After analyzing the data, it appears that multiple cold shocks slightly decreases the meffBlue expression levels in the experimental group. However, the control group appeared to show a similar trend for the protein expression being measured at a wavelength 592. Therefore, no conclusive quantitative can show how the TSA responds to multiple cold shocks.

Stanford Location

Barcode #: 0133010781 & 0133011771
Plasmid Name: Temperature Sensor/Actuator
Antibiotic Resistance: Ampicillin
DNA 2.0 Gene #: 193460
Organims Expressed In: E.coli
Sensor/Actuator: Both

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