Part:BBa_K569014
SCP+ToxR+FGFR
This part constitutively produces the ToxR+FGFR fusion protein. ToxR+FGFR dimerizes in response to FGF and can then activate the ctx promoter from the bacteria Vibrio cholera. ToxR by itself is a transcription factor, while FGFR is a immune system sensor.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 7
Illegal NheI site found at 30 - 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 1266
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 844
Illegal NgoMIV site found at 1435 - 1000COMPATIBLE WITH RFC[1000]
Preliminary Experiments
We transformed SCP+ToxR+FGFR (BBa_K569014) and ctx+GFP (BBa_J07011) into DH5a E.coli cells using two antibiotics: chloramphenicol and kanamycin. We selected a single colony and grew it in LB with chloramphenicol and kanamycin overnight at 37C and 220rpm. We made a 1:20 dilution of the sample and then grew it to a predetermined O.D. When the O.D. was reached induced with various amounts of heparin and fibroblast growth factor (FGF)and let it grow. We took measurements using fluorescence microscopy.
All images were taken with Olympus IX81 automated inverted microscope specially equipped for live cell imaging. The filter set we used is: 470/40x (excitation) and 525/50m (emission) for GFP. Data collection and processing was performed by the SlideBook software.
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