Defense Date

8-8-2007

Graduation Date

2007

Availability

Immediate Access

Submission Type

thesis

Degree Name

MS

Department

Biological Sciences

Committee Chair

Joseph R. McCormick

Committee Member

John F. Stolz

Committee Member

Nancy Trun

Keywords

cell division, divisome, epitope tag, protein complexes, protein-protein interactions, pull down assay

Abstract

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.

Format

PDF

Language

English

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