Part:BBa_K3788025
pelB_sfGFP
BBa_K3788025 is coding for pelB and sfGFP. pelB is a leader sequence, it allows the protein to join the periplasm via the SEC system (unfolded proteins). In this contruction sfGFP permit to understand the mechanism, this protein can at the end be replaced by an other protein. sfGFP is supposed to be address to the preriplasm thanks to pelB leader and get his folded form into the periplasm.
Cell visualization with confocal microscope
We wanted to determine whether the fluorescence is located in the periplasm or in the cytoplasm, so we used a confocal microscope to visualize our cells.
We transformed E.Coli MG1655 strains with an empty J04500 vector or with the BBa_K3788025 recombinant plasmid. We then prepared 3mL 2YT growing medium with chloramphenicol and then put MG1655 containing either J04500 or BBa_K3788025 plasmids.
After 3 hours of incubation at 37°C, we prepared the plates and visualized our cells
Figure : Visualisation of MG1655 E.coli strains containing J04500
Figure : Visualisation of MG1655 E.coli strains containing BBa_K3788025
We can see that bacteria containing J04500 do not emit light, contrary to the ones containing J04500_PelBsfGFP, which are green under a confocal microscope.
However, we are not able to determine whether the light is located at the surface of the bacteria or if it is located in all the cells.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 54
- 1000INCOMPATIBLE WITH RFC[1000]Illegal SapI.rc site found at 79
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