Optimization of Parameters for the Construction of a Sensing System to Measure the Hydrolysis of ß-Lactam Antibiotics

ASU Author/Contributor (non-ASU co-authors, if there are any, appear on document)
Nicholas David McGuire (Creator)
Institution
Appalachian State University (ASU )
Web Site: https://library.appstate.edu/
Advisor
Libby Puckett,

Abstract: Bacterial resistance to antibiotics is a problem that has developed over the last 75 yearsdue to over-prescribing and improper usage of these drugs. Through these processes, bacteriahave developed and spread the gene for an enzyme known as ß-lactamase, which catalyzes thehydrolysis of the ß-lactam ring in many penicillins, rendering them ineffective. Accordingly, thepurpose of this research is to create a sensing system to measure the hydrolysis of ß-lactamantibiotics using the enzyme, ß-lactamase. To accomplish this, the gene for ß-lactamase can befused with the gene for a fluorescent protein known as enhanced green fluorescent protein(EGFP), which will decrease its fluorescence upon the local pH change generated by thecatalysis of a ß-lactam ring. While previous research in this area has been successful in thedevelopment of an in vitro fusion of the genes encoding for ß-lactamase and EGFP, presentresearch is focused on separately incorporating the individual genes for EGFP and ß-lactamase inpFLAG-MAC expression vectors to verify the local pH theory and to create an in vivo protein,thus creating a whole cell sensing system.

Additional Information

Publication
Honors Project
McGuire, N.D. (2015). Optimization of Parameters for the Construction of a Sensing System to Measure the Hydrolysis of ß-Lactam Antibiotics. Unpublished honors thesis. Appalachian State University, Boone, NC.
Language: English
Date: 2015

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