4th Annual WML Academic Symposium

Diabetes is a disease characterized by dysfunctional blood sugar regulation due to errors in glucose metabolism associated with the insulin signaling pathway. Type I Diabetes is a form of diabetes where genetic errors prevent sufficient insulin from being produced. Beta cells, the pancreatic cells responsible for insulin production, don’t develop in patients with type I Diabetes. Advances in genetic therapy options could yield a cure for Type I Diabetes.

To express insulin, general transcription factors and pancreatic transcription factors bind to the promoter of the insulin gene to initiate transcription. Pdx1, a general transcription factor, and MafA, a pancreatic specific transcription factor, have adjacent DNA binding sites on the insulin gene promoter. In vivo, these transcription factors form complexes prior to transcription, and these complexes are known to synergistically enhance insulin transcription.

The gene sequences of Pdx1 and MafA were inserted into an expression vector, separated by a seventy-eight nucleotide linker. The transcription factor fusion protein was expressed in E. coli by IPTG induction, and aliquots of whole cell lysates were taken at two-hour time intervals. The whole cell lysates were run on a polyacrylamide gel to test for increased expression of the fusion protein over time.