Difference between revisions of "Part:BBa K3788025"

Revision as of 15:10, 21 October 2021

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