Ruff, Kara

Project title: Proof of principle: targeting of GFP to Dictyostelium mitochondrial matrix

Department: Priority Initiative - Biology

Faculty supervisor: Dr. Ryan Gawryluk

"An emerging protistan model of mitochondrial biology is the slime mold, Dictyostelium. As an amoebozoan, Dictyostelium occupies a key evolutionary position (Amoebozoa is sister to animals + fungi), and its complex life cycle – involving both unicellular and multicellular stages – may help elucidate the mitochondrion’s role in the evolution of multicellularity. However, the lack of a well-curated Dictyostelium mitochondrial proteome is an impediment to its utility in mitochondrial research. The Gawryluk laboratory aims to characterize the mitochondrial proteome in Dictyostelium by targeting a biotinylating protein BioID to the mitochondrial matrix, subsequently purifying the proteins, and identifying them by mass spectrometry. This project aims to develop a necessary control by demonstrating that the all steps in the protocol are effective in localizing a protein to the matrix. I will extract RNA from Dictyostelium to attain the N-terminal targeting peptide, ligate it into a GFP plasmid, transform Dictyostelium with the plasmid, and co-localize GFP with MitoTracker."