Document Type

Paper- Restricted to Campus Access

Publication Date

4-22-2021

Faculty Mentor

Samantha Wilner

Abstract

4DIU is a protein that does not currently have a known function. We aim to add to scientific knowledge through our analysis of 4DIU. Our research contains a combination of in-lab and online techniques in order to collect information about 4DIU. E.Coli were transformed with the expression vector UN2 to create protein. Ampicillin resistant cultures were grown and used for the induced expression of the protein in LB media. Then the protein expression was induced. Nickel resin chromatography was used to purify the protein and was successful once imidazole was added as a competitive inhibitor to elute 4DIU from the column. An SDS-PAGE gel electrophoresis analysis showed the large presence of protein at the proper kDa of 28. In collaboration with Dr. Roberts, we determined 4DIU has a catalytic triad of Ser19, His222, and Asp 192. It most closely matches the carboxylesterase precursor 1THQ. Bradford Assay was used to find the protein concentration. Enzymatic and Kinetic activities of 4DIU were observed. Our research of 4DIU contributes to the understanding of the protein's structure and function.

Comments

Presented as part of the Ursinus College Celebration of Student Achievement (CoSA) held April 22, 2021.

The downloadable file is a poster presentation with audio commentary with a run time of 5:57.

Conducted as lab research in Biochemistry 452.

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