The accomplishment of our project has been a long process over the course of almost a year. The experiments we have performed, lead to the results explained below. More in-depth details about these results are explained in the Experiments page.
The goal of our project was to create a more universal bacterial tracking system
than
the one developed by Gregor et al. (2018), based on the ilux operon. We wanted to:
- make ilux compatible with the Biobrick format;
- place ilux under the control of a constitutive promoter;
- avoid an energetic cost for the bacterium;
- put the system on a conjugative plasmid with a replication origin for a broad spectrum of hosts.
Our fiatlux operon is a mutated version of ilux. Indeed, we needed it in a BioBrick format and wanted to make it adaptable to many strains of bacteria so that we could track as many bacteria as possible in situ. And we did it!
Here are two timelapse videos of FIAT LUX-containing bacteria, propagating and growing in situ and in Petri dishes.
- Left: D.solani containing pSEVA531-fiatlux plasmid, infecting chicory leaves. Their propagation can be followed in real time throughout the chicory tissues. The pictures are taken every 30 minutes, during 24 hours.
- Right: D.solani and E.coli DH5alpha containing the plasmids (pSEVA521-fiatlux or pSEVA531-fiatlux) on Petri dishes in LB medium with different concentrations of tetracycline (0,5 and 10 µg/mL). The pictures are taken every 30 minutes, during 24 hours.
Here is the roadmap of all our major steps to achieve our goal! You can find the details for all plasmids used in the Material page.
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