Team:Sorbonne U Paris/Demonstrate

HiBiT Tag

We were able to detect proteins with the HiBiT tag in a very sensitive way. Indeed, for the tagged GFP, the expression was too low to detect the GFP fluorescence, but we were still able to detect the protein thanks to the HiBiT tag.
We quantified precisely the expression of the protein with HiBiT tag, helping us to choose high expressing clone. In our assay, we were able to identify constitutive expressing cells and thus, we could investigate the modification of the lipid profile in ammonium condition.
In our project, the HiBiT detection system allowed us to detect easily and quickly the exogenous enzymes. Without this system, we would have needed a heavier detection method, like the ones based on antibodies (e.g. Western Blot, ELISA). The HiBiT detection system allows us to screen lots of clones at the same time to identify expressing clones as well as clones with high level of expression.




To conclude, we introduced the HiBiT technologies into the C. Reinhardtii Modular Cloning kit, providing a powerful tool to detect and quantify proteins.

Modification of lipid metabolism

We were able to modify the lipid profile of C. Reinhardtii by adding exogenous enzyme from oil palm, proving that some enzymes from palm oil can affect C. Reinhardtii metabolism, meaning that first, they are targeted at the correct compartment, and second that they recognize their substrate.




Even though it is very preliminary results, it could mean that the addition of several enzymes, as we first planed, could allow the lipid profile of C. Reinhardtii to tend towards the palm oil lipid profile, as well as increasing the production of triglycerides in those cells.
Thus, our project showed the possibility of tweaking the lipid metabolism of C. Reinhardtii to modify the lipid profile as wanted by adding genes coding for metabolism enzymes from other organisms.