2019 Student Research Conference:
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

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