Primary Submission Contact

Student: Alyson Kneusel Faculty: Laura Rowe

Faculty Sponsor

Dr. Thomas Goyne

Faculty Sponsor Email Address

thomas.goyne@valpo.edu

College

Arts and Sciences

Department/Program

Chemistry Department

Document Type

Poster Presentation

Date

Fall 10-28-2016

Abstract

The goal of this work is to identify unnatural amino acids (UAAs) that can be incorporated into proteins by E. coli using four orthogonal aminoacyl tRNA synthetase/suppressor tRNA (aaRS/tRNA) pairs. A fluorescence screening assay was used to test the incorporation of thirteen UAAs into proteins using four aaRS/tRNA pairs that had been previously shown the possess varying degrees of promiscuity. The screen utilized a mutant green fluorescent protein (GFP) that included the amber stop codon within the open reading frame. Multiple results demonstrated that aaRS/tRNA pairs are able to bind other UAAs to a level greater than the UAA designed to match that synthetase, yet all UAA mutants had significantly lower incorporation than the wild type GFP control (UAA absent). Correlation of the relative incorporation levels versus the UAA structures provides a basis for understanding the shape and flexibility of the synthetase binding pocket and thus provides experimental data that can be used to evaluate molecular modeling results.

Biographical Information about Author(s)

Alyson Kneusel is a Junior Chemistry and Biology double major from Tampa, Florida. She hopes to go to graduate school and pursue a career in Biomedical research. This project aligns well with her interests as it has biochemical and bio-analytic focuses. She began working on the project during summer 2016 and has continued work on it throughout the fall semester.

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