Part:BBa_K3788025:Experience

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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.

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