Date of Graduation
Master of Science in Cell & Molecular Biology (MS)
Douglas D. Rhoads
Second Committee Member
Pure sciences; Biological sciences; Amylin; E. coli; Islet amyloid polypeptide; Recombinant technology
Islet amyloid polypeptide (IAPP) also called amylin is an amyloid-forming protein; IAPP is a proteinaceous hormone that comprises 37 amino acid residues. It is secreted along with insulin from the pancreatic β-cells to help it regulate the uptake and removal of glucose in the bloodstream. IAPP has been observed in the amyloid deposits found in pancreatic β-cells of most patients suffering from type II diabetes mellitus. This research project aims at producing recombinant amylin peptide. To achieve this goal, we used the pBAD plasmid vector which we introduced into Escherichia coli to express the peptide. Although the vector was successfully introduced into E. coli, production of the amylin protein was not detected under a variety of different expression conditions. Examination of the RNA produced from the E coli showed expression of the amylin RNA which indicates that the protein is most likely degraded. The degradation may have resulted from the fact that the peptide in question is a small exogenous toxic peptide and as a result may have been degraded by cytoplasmic proteases to protect the cell. One way that this degradation could be overcome is to attach the protein to an endogenous fusion peptide, such as the maltose-binding protein, which could provide protection against proteases.
Duverna, Emilio, "Production and Characterization of Islet Amyloid Polypeptide Using Recombinant Technology" (2015). Theses and Dissertations. 1196.