In the bodies of people with diabetes, the ability to recognize and respond to glucose concentrations in the blood has been compromised. As a result, glucose accumulates to dangerous levels. High blood glucose concentrations can cause irreversible damage to critical organs, impairing their functionality. With parts from the iGEM registry, our team has created a osmolarity controlled promoter linked to a yellow fluorescence production gene in E. coli. Osmolarity is the measure of solutes that contribute to the osmotic pressure of a solution, so as the glucose concentration goes up the osmolarity changes. The concentrations of glucose to which the promoter responds are initially unknown. Once the concentrations are known, the promoter can be mutated so that it will be activated by varying concentrations of glucose and be used as a glucose sensor for people with diabetes. In the future, an insulin gene could be added to this system for use in insulin pumps, where specific glucose levels trigger insulin production in E. coli.

To check results and separate plasmids and genes, the team will use Gel electrophoresis. For instance, after the initial digests in which Omp-R, eYFP, and RBS have all been cut in the appropriate places, the team will run a gel first: to make sure that the digestion worked as planned, and second: to separate the various pieces from their original plasmids. We will also be useing cultures with Ampicillin to make sure that the ligations are viable and that they have the right genes. Each Digestion will be followed by a Gel and each ligation will be followed by transformation. To veiw the results of these experiments, visit the Notebook tab.