Composite

Part:BBa_K3594013

Designed by: Runhan Luo   Group: iGEM20_SHSBNU_China   (2020-10-23)
Revision as of 18:06, 23 October 2020 by GraceLuo (Talk | contribs)


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.

T--SHSBNU China--the process of translation in different temperature.png

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.

T--SHSBNU China--ways to calculate normalized fluorescence.jpeg
File:T--SHSBNU China--the process we do to test the function of BBa K3594013.png
the process we do to test the function of BBa K3594013


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal XhoI site found at 450
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


[edit]
Categories
Parameters
None