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Presentation Type

Presentation

Full Name of Faculty Mentor

Daniel Williams, Biology

Major

Biology

Presentation Abstract

Bacteriophages are ubiquitous viruses containing diverse genomes. Many phage genomes have been bioinformatically annotated; however, many genes lack wet-bench functional characterization. Elucidating individual phage gene function on host growth and the phage-host protein interactions causing such phenotypes allows for the exploration of novel antibacterial therapies within the context of phage-host biology. We aim to systematically characterize the genome of the temperate mycobacteriophage Phayonce through the investigation of individual gene expression on host cell growth and the phage-host protein interactions underscoring their cytotoxicity. A library of inducible expression vectors was generated containing each of Phayonce’s 77 genes. Individual plasmids were transformed into the host cell Mycobacterium smegmatis. When induced on selective media, the resulting host colony phenotype was observed, with 29 genes exhibiting cytotoxic activity. Of these genes, genes 41 and 64, which lack a characterized function, showed near-total inhibition of colony formation. To identify host proteins interacting with these genes, we performed a bacterial two-hybrid screen and isolated numerous host protein fragments of possible interaction partners. Overall, Phayonce encodes numerous, poorly characterized cytotoxic genes. Given their effect on host colony formation, these genes represent a reservoir of potential candidates to be exploited in antibacterial therapeutic development. Since novel genes 41 and 64 exhibit extreme cytotoxicity, they represent prime examples of genes that can be utilized as such. Isolated phage-host protein interaction candidates will be verified and subsequently sequenced to identify the host interaction partners mediating the cytotoxic phenotype. Once identified, these interaction partners can also be exploited in antibacterial therapeutics.

Start Date

11-4-2023 8:00 AM

End Date

11-4-2023 8:20 AM

Disciplines

Biology

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Apr 11th, 8:00 AM Apr 11th, 8:20 AM

Systematic Characterization of Mycobacteriophage Gene Function on Bacterial Cell Growth

Bacteriophages are ubiquitous viruses containing diverse genomes. Many phage genomes have been bioinformatically annotated; however, many genes lack wet-bench functional characterization. Elucidating individual phage gene function on host growth and the phage-host protein interactions causing such phenotypes allows for the exploration of novel antibacterial therapies within the context of phage-host biology. We aim to systematically characterize the genome of the temperate mycobacteriophage Phayonce through the investigation of individual gene expression on host cell growth and the phage-host protein interactions underscoring their cytotoxicity. A library of inducible expression vectors was generated containing each of Phayonce’s 77 genes. Individual plasmids were transformed into the host cell Mycobacterium smegmatis. When induced on selective media, the resulting host colony phenotype was observed, with 29 genes exhibiting cytotoxic activity. Of these genes, genes 41 and 64, which lack a characterized function, showed near-total inhibition of colony formation. To identify host proteins interacting with these genes, we performed a bacterial two-hybrid screen and isolated numerous host protein fragments of possible interaction partners. Overall, Phayonce encodes numerous, poorly characterized cytotoxic genes. Given their effect on host colony formation, these genes represent a reservoir of potential candidates to be exploited in antibacterial therapeutic development. Since novel genes 41 and 64 exhibit extreme cytotoxicity, they represent prime examples of genes that can be utilized as such. Isolated phage-host protein interaction candidates will be verified and subsequently sequenced to identify the host interaction partners mediating the cytotoxic phenotype. Once identified, these interaction partners can also be exploited in antibacterial therapeutics.