Construction of a Novel E. coli and S. aureus Shuttle Vector and Promoter-Nanoluciferase Fusion Reporter Plasmid and Initial Characterization of SaPI1 and Phage 80a Packaging Interactions
Location
Sydnor Performance Hall, Schewel Hall
Access Type
Campus Access Only
Presentation Type
Oral presentation
Entry Number
82
Start Date
4-16-2026 2:00 PM
End Date
4-16-2026 2:15 PM
School
School of Liberal Arts and Sciences
Department
Biology
Abstract
Staphylococcus aureus is a common multidrug-resistant bacteria that causes many clinical diseases such as meningitis, toxic shock syndrome, and urinary tract infections. In response to pervasive antibiotic resistance, this project aims to investigate the interaction between S. aureus' cellular machinery and chromosomal components as a potential therapeutic approach. In addition to its genome, S. aureus often contains integrated bacteriophages (prophages) and pathogenicity islands. Prophages are dormant until induced, and only then do they hijack the cell's machinery to construct virions and kill the host cell via lysis. We have preliminary evidence that the pathogenicity island SaPI1 can interfere with prophage gene expression, encoding a potential novel transcription factor that activates an uncharacterized promoter in the phage genome. To identify this secondary promoter, we will construct a series of promoter-fusion plasmids. Current plasmid options and reporter methods are limited in S. aureus; therefore, our first objective was to construct a novel shuttle vector with nanoluciferase fusion. We have successfully constructed two plasmids, pBASE1 and pBASE2 (pBASE1 with a nanoluciferase gene added), and will now assess their transmission efficiency, effects on host cell growth, and the functionality of the promoter fusion using a wildtype S. aureus promoter, a constitutively overexpressing promoter, and a silent promoter mutant. Characterizing the function of these novel plasmids will allow us to identify the exact location of the proposed secondary promoter in the phage genome, providing new insight into the interaction between bacteriophages and pathogenicity islands during host cell lysis.
Primary Faculty Mentor(s)
Dr. Jamie L. Brooks
Primary Faculty Mentor(s) Department
Biology
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Construction of a Novel E. coli and S. aureus Shuttle Vector and Promoter-Nanoluciferase Fusion Reporter Plasmid and Initial Characterization of SaPI1 and Phage 80a Packaging Interactions
Sydnor Performance Hall, Schewel Hall
Staphylococcus aureus is a common multidrug-resistant bacteria that causes many clinical diseases such as meningitis, toxic shock syndrome, and urinary tract infections. In response to pervasive antibiotic resistance, this project aims to investigate the interaction between S. aureus' cellular machinery and chromosomal components as a potential therapeutic approach. In addition to its genome, S. aureus often contains integrated bacteriophages (prophages) and pathogenicity islands. Prophages are dormant until induced, and only then do they hijack the cell's machinery to construct virions and kill the host cell via lysis. We have preliminary evidence that the pathogenicity island SaPI1 can interfere with prophage gene expression, encoding a potential novel transcription factor that activates an uncharacterized promoter in the phage genome. To identify this secondary promoter, we will construct a series of promoter-fusion plasmids. Current plasmid options and reporter methods are limited in S. aureus; therefore, our first objective was to construct a novel shuttle vector with nanoluciferase fusion. We have successfully constructed two plasmids, pBASE1 and pBASE2 (pBASE1 with a nanoluciferase gene added), and will now assess their transmission efficiency, effects on host cell growth, and the functionality of the promoter fusion using a wildtype S. aureus promoter, a constitutively overexpressing promoter, and a silent promoter mutant. Characterizing the function of these novel plasmids will allow us to identify the exact location of the proposed secondary promoter in the phage genome, providing new insight into the interaction between bacteriophages and pathogenicity islands during host cell lysis.