Title

Expression and Purification of FepI Involved in Iron Regulation in S. pombe

School Name

Governor's School for Science and Mathematics

Grade Level

12th Grade

Presentation Topic

Microbiology

Presentation Type

Mentored

Abstract

The aim of this project was to purify and characterize the His-tagged Fep1 protein through the use of bacterial expression plasmids with the aid of biochemical and spectroscopic tools. Schizosaccharomyces pombe is a useful model eukaryotic system for studying regulation of iron homeostasis. Results from both S. pombe and Saccharomyces cerevisiae have shed light on the key players of this complex regulation. Proteins like monothiol glutaredoxins (Grxs) are present in both types of yeasts yet display different regulatory functions. Since S. pombe and S. cerevisiae share a common ancestor, the lab hypothesized that extrapolating the knowledge obtained from S. cerevisiae to S. pombe would allow the lab to fully understand the function of monothiol glutaredoxins in S. pombe. The lab planned to obtain these results through the purification and characterization of Fep1 (from S. pombe) in vitro using the bacterial expression plasmids pET-28a(+)6xHIS-Fep1(2-564)WT and pET28(+)Fep1-6xHIS(1-564)WT.

Location

Neville 221

Start Date

4-14-2018 9:00 AM

Presentation Format

Oral and Written

COinS
 
Apr 14th, 9:00 AM

Expression and Purification of FepI Involved in Iron Regulation in S. pombe

Neville 221

The aim of this project was to purify and characterize the His-tagged Fep1 protein through the use of bacterial expression plasmids with the aid of biochemical and spectroscopic tools. Schizosaccharomyces pombe is a useful model eukaryotic system for studying regulation of iron homeostasis. Results from both S. pombe and Saccharomyces cerevisiae have shed light on the key players of this complex regulation. Proteins like monothiol glutaredoxins (Grxs) are present in both types of yeasts yet display different regulatory functions. Since S. pombe and S. cerevisiae share a common ancestor, the lab hypothesized that extrapolating the knowledge obtained from S. cerevisiae to S. pombe would allow the lab to fully understand the function of monothiol glutaredoxins in S. pombe. The lab planned to obtain these results through the purification and characterization of Fep1 (from S. pombe) in vitro using the bacterial expression plasmids pET-28a(+)6xHIS-Fep1(2-564)WT and pET28(+)Fep1-6xHIS(1-564)WT.