2025 Undergraduate Research Showcase

Molecular Cloning of Staphylococcus aureus Alpha Toxin as an Antigen for Vaccine Development

Document Type

Student Presentation

Presentation Date

4-15-2025

Faculty Sponsor

Dr. Juliette Tinker and Dr. Elise Overgaard

Abstract

Staphylococcus aureus is a gram positive bacterium known for causing a wide variety of dangerous infections in humans, as well as a disease known as mastitis in dairy cows. The development of a vaccine to target S. aureus bovine mastitis would be a more effective and less expensive treatment for the disease than culling or antibiotics. The S. aureus hla gene codes for alpha toxin (also known as hemolysin), a protein that acts as a virulence factor for S. aureus related infections. To begin production of the hemolysin protein for use as an antigen in the vaccine, PCR with agarose gel electrophoresis was used to isolate and amplify the hla gene from S. aureus strain NCTC 8325 / PS 47. The PCR product was then ligated into two different vectors (pAJW003 and pARLDR19), and the resulting plasmids (pRK001 and pRK002, respectively) were each transformed into TE1 E. coli cells. pRK001 transformed cells grew successfully on Luria-Bertani + chloramphenicol + glucose (LB+cm+glu) plates, and PCR with gel electrophoresis was used again with the same primers as before to confirm successful transformation. pRK001 colonies were saved as glycerol stocks for future purification of the hemolysin protein as part of the vaccine. pRK002 transformed cells did not present any growth on LB+cm+glu plates, and are currently under further experimentation.

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