Bayer School of Natural and Environmental Sciences
Joseph R. McCormick
John F. Stolz
cell division, divisome, epitope tag, protein complexes, protein-protein interactions, pull down assay
In Streptomyces coelicolor, at least six proteins are involved in the process of cell division: FtsZ, FtsQ, FtsL, DivIC, FtsW, and FtsI. The purpose of this study was to biochemically analyze putative cell division protein interactions to complement and possibly expand upon previous genetic studies. To accomplish this, S. coelicolor strains expressing epitope-tagged cell division proteins were created and pilot experiments using a Ni-NTA pull down assay were performed. His8-FtsZ was found to be stable in vivo; however, many fusion proteins were found to be nonfunctional. The Ni-NTA pull down assay worked well to isolate His8-FtsZ, but other fusion proteins could not be visualized by western blot analysis due to low levels of expression. The antibodies used in western blot analyses were found to be problematic due to high background and nonreactivity. Attempts to optimize these assays were performed; however, further optimization is needed to continue this study.
Kotun, A. (2007). Biochemical Analysis of Cell Division Protein Complexes in Streptomyces coelicolor (Master's thesis, Duquesne University). Retrieved from https://dsc.duq.edu/etd/772