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.
Recommended Citation
Sanyang, Marie, "Expression and Purification of FepI Involved in Iron Regulation in S. pombe" (2018). South Carolina Junior Academy of Science. 69.
https://scholarexchange.furman.edu/scjas/2018/all/69
Location
Neville 221
Start Date
4-14-2018 9:00 AM
Presentation Format
Oral and Written
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.
