FRES(H): Fruit ripeness ethylene biosensor (hopefully)
Biosensors utilise the genetic circuits found in nature for sensing of particular substrates. In our case, we turned to ethylene-metabolising bacteria, specifically a local Australian strain of Mycobacterium known as Mycobacterium NBB4.
SENSE: The regulatory genes behind ethylene metabolism in NBB4 are almost completely novel . Based on sequence homology and structural protein analysis, we've come up with two strong candidates - genes called EtnR1 and EtnR2, which may interact with a putative promoter EtnP. Therefore, we had to check expression and characterise these regulatory components first!
EXPRESS: If we want to use the regulatory aspects of NBB4 ethylene metabolism in a biosensor, we need to incorporate a reporter gene with strong, detectable output into our circuit. Generating such a reporter gene was the focus of EXPRESS, where we used error prone PCR on an existing blue iGEM chromoprotein amilCP to generate amilCP mutants with altered spectral characteristics.
KEEP FRESH: Putting SENSE and EXPRESS biocircuitry together, we can construct a usable biosensor with applications in cold shipping containers, supermarket store rooms and storage warehouses. We focused on the physical design aspects of a biosensor to bring to market, and performed detailed costings in a business plan for our biosensor.