Part:BBa_K3594013
Heat Responsive Fluorescence Emitting Device
This composite plasmid is used as the reporter of our heat-responsive suicide switch. The expression of sfGFP gene is controlled by an RNA-based thermal sensor, which was constructed by Jilin_China 2018 team.
When the temperature is higher than 37℃, the ribosome starts to work and the downstream gene sequence can be expressed. Specifically, in this part, the fluorescence would be emitted.
Characterization
Each component of bacteria with BBa_K3594013 was divided into 3 biological controls, which were placed at 30℃, 33℃, 37℃, 42℃ with the negative control, positive control and blank control LB, and shaking at 220 rpm. We took samples at 0h, 4h, 8h and 20h respectively, used a microplate reader for spectrophotometer and fluorescence test, and recorded the data in OD600 and Fluorescence 485/510 (Excitation/Emission). We calculated the value of Normalized Fluorescence changes with the equation referenced from Jilin_2018 iGEM team at different temperatures and different time.
We expressed the data are a figure:
Normalized Fluorescence of BBa_K3594013 changes with time at different temperatures. The abscissa represents the time, and the ordinate is the Normalized Fluorescence value. From the image, it can be seen that the Normalized Fluorescence increases with the increase of time. The value of Normalized Fluorescence in 42 ℃ increases most obviously, which proves that the suicide switch is effective at high temperatures.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal XhoI site found at 450
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
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