Speaker: Annie Mancha
Candidate for Bachelor of Science in Biochemistry
Time: 6:00 PM
Place: Trustee Hall 118
Supervisor: Dr. Mary Kopecki-Fjetland
Title: Partial Purification and Characterization of a Potential Rubredoxin from Anabaena 7120

Abstract: Rubredoxins are small iron-sulfur proteins believed to be involved in electron transport. They are defined by the iron-binding cysteine residues, which are ubiquitously conserved. Prior research showed that a gene in Anabaena 7120 encodes a protein displaying similarity to rubredoxins, which has been termed anaredoxin. The sequence homology to known rubredoxins is only seventeen percent, but the iron binding cysteine residues align perfectly. We hypothesize that the anaredoxin is a rubredoxin. In order to prove this hypothesis, anaredoxin must be expressed, purified, and characterized. In previous work, the potential anaredoxin protein appeared to be overexpressed upon addition of the inducer IPTG to a bacterial cell culture grown to an A600 of 1.0 at 37 degrees Celsius. In order to more clearly verify this potential overexpression, we are partially purifying anaredoxin using gel filtration chromatography. Samples of the partially purified protein will then be analyzed using SDS-PAGE and spectrophotometry. Future studies include subcloning the gene into an expression system to produce a tagged fusion protein for subsequent characterization.