32nd Annual Student Research Conference
Expression and Purification of DBF Based on Optical Density
Ryan P. Poland* and Mary C. Butler
Dr. Cassidy Dobson, Faculty Mentor
Expression and Purification of DBF Based on Optical Density
Ryan Poland and Mary Butler
Faculty Mentor: Dr. Cassidy Dobson
Department of Chemistry, Truman State University
Background: Cataracts, the leading cause of blindness worldwide, are caused by aggregations of disulfide bonds between γ crystallins in the eye. Previous research suggests that Disulfide Bond Forming Enzyme (DBF) can rearrange the covalent sulfur-sulfur bonds present in cataracts.
Methods: Recombinant expression of DBF was performed in E. coli using pRHAM or pET28-a vectors. Protein induction was carried out at different optical densities to observe optimized protein expression.
Results: Based on data collected, the optimal OD to induce the cell culture is closer to 0.4 than 1.0 due to results suggesting more purified protein.
Discussion: Determining the optimal expression and purification conditions would increase efficiency and productivity in the laboratory. The data collected so far indicates that lower OD leads to increase protein production, but further testing will be conducted.
Keywords: Expression, Purification, Optical Density, Cataracts, Disulfide Bonds, γ crystallins, Protein, Aggregations
Topic(s):Biochemistry and Molecular Biology
Chemistry
Biology
Presentation Type: Poster
Session: 9-2
Location: SUB GEO
Time: 3:00