2024 Undergraduate Research Showcase

Eat the Raw Cookie Dough: Salmonella Vaccine Development by Exploring Salmonella Toxins

Document Type

Student Presentation

Presentation Date

4-19-2024

Faculty Sponsor

Dr. Juliette Tinker

Abstract

Salmonella is a highly antibiotic-resistant Gram-negative bacteria that is the most reported bacterial food-borne disease. Despite increases in awareness and sanitation, the incidence of Salmonella has continued to increase. A vaccine to combat this important pathogen in agriculturally important animals would help to protect humans. AB5 toxins are key bacterial virulence factors and have been used as vaccine antigens. We have previously characterized an AB5 toxin of S. Typhimurium, called ArtAB, to better understand its role in pathogenesis. We hypothesized that other AB5 toxins can be found using Salmonella whole genome sequence searches. We identified a homologous toxin with 31% identity to ArtAB in S. Montevideo (PltC). Using PCR, we have also identified PltC in isolates of S. Typhimurium and cloned it into a plasmid for expression in E.coli. A similar process has been used to identify a toxin from Diarizonae that has two B-subunits to clone into E. Coli. Protein purification using D-galactose or Fetuin affinity was not successful to purify the pltC A and B subunits, and other methods are currently being explored including cloning the B subunit alone. In addition, we have tested the DNA from bovine fecal samples to determine if the artAB gene is present and will test these samples for pltC in the future. Western blot was used to test for an immune response, but the process was unsuccessful. This work will support future studies involving antigen and antibody testing with the goal of animal studies and clinical trials to develop a novel bovine Salmonella vaccine.

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