Team:Aix-Marseille/Results
Wetlab
Endochitinase
The endochitinase is a native protein produced by Beauveria bassiana an enthomopathogenic fungus. We wanted to overproduce (in E. coli) and extract it to enhance the killing efficieny and speed of the fungus. We succesfully produced, purified a functional endochitinase (|BBa_K2718022)
Pheromones production
The aim was to produce two types of pheromones: DMTS/DMDS and benzaldehyde/benzylalcohol. We did some research on the two molecules to get the biosynthesis pathways to produce them in E. coli. Firslty, we needed to add the three enzymes implicated in the biosynthesis of benzaldehyde/benzylalcohol: MdlB, MdlC and Hmas. We successfully cloned the Hmas gene (BBa_K2718011) and produced the equivalent protein. This enabled the production of the first metabolic intermediate (S-Mandelate) of the pathway. Secondly, we were able to clone the methionine-γ-lyase gene (BBa_K2718006) and produce the equivalent protein. This enable the achievement of a first goal: the production of the DMTS/DMDS pheromone.
Trap tests
Model
We decided to model the deployment of our bed bug trap in a realistic environment, to understand how it was likely to work and what parameters might be important. This task involved a number of different modeling tasks, that posed different problems: modelling the diffusion of pheromones in the room coming both from a natural bed bug nest and the traps that we plan to deploy; modeling the movement of bed bugs influenced both by the pheromone field and their nest; and modeling the fungal epidemic that we plan to induce in the bed bug population. Though the model is complex and includes many parameters, it has already allowed us to draw several conclusions, and as the model is improved and the parameters are refined other conclusions will follow. For instance, using realistic diffusion parameters it is clear that the pheromone field is relatively rapidly established and so the precise nature and concentration of pheromones is probably not critical. In contrast, the delay between infection and death of bed bugs is critical for ensuring the eradication of the nest. Our modeling thus helps understand critical aspects of the proposed design.
Interlab
The InterLab study has been developing a robust measurement procedure for green fluorescent protein (GFP) over the last several years. GFP was chosen as the measurement marker for this study since it's one of the most used markers in synthetic biology and, as a result, most laboratories are equipped to measure this protein. This year, we helped in answering the following question: Can we reduce lab-to-lab variability in fluorescence measurements by normalizing to absolute cell count or colony-forming units (CFUs) instead of OD?