Generating a Gene Library of Bacteriophage Phayonce
Presentation Type
Presentation
Full Name of Faculty Mentor
Daniel Williams, Biology
Major
Biology
Presentation Abstract
Due to their toxicity for bacterial hosts, bacteriophages have an emerging importance in treating medically significant bacterial species. Generating a gene library of Phayonce, a bacteriophage that infects Mycobacterium smegmatis, allows for systematic analysis of individual function and possible cytotoxic effects on host cells. This work aims to generate a library that contains each of Phayonce’s 77 genes in an inducible expression vector. Individually cloned genes can then be introduced into M. smegmatis host cells for subsequent analysis of gene function. Each Phayonce gene was amplified by PCR using gene specific primers. Amplified genes were then assembled into the pExTra plasmid, which contains a tetracycline inducible promoter to drive Phayonce gene expression. After assembly and transformation into E.coli, colonies containing putative plasmids were analyzed by PCR to verify the insert. All 77 genes of Phayonce’s genome were successfully cloned into pExTra and subsequent sequencing established they are error-free. Generating a complete gene library of the bacteriophage Phayonce has allowed for all 77 expression vectors to be used in phenotypic analysis of toxicity of each Phayonce gene. Toxicity of Phayonce genes on host M. smegmatis may have medically significant implications on bacteriophage therapies for bacterial species that have shown to be resistant to treatment.
Start Date
13-4-2023 2:00 PM
End Date
13-4-2023 2:20 PM
Disciplines
Biology
Recommended Citation
Thomas, Kyla, "Generating a Gene Library of Bacteriophage Phayonce" (2023). Undergraduate Research Competition. 105.
https://digitalcommons.coastal.edu/ugrc/2023/fullconference/105
Generating a Gene Library of Bacteriophage Phayonce
Due to their toxicity for bacterial hosts, bacteriophages have an emerging importance in treating medically significant bacterial species. Generating a gene library of Phayonce, a bacteriophage that infects Mycobacterium smegmatis, allows for systematic analysis of individual function and possible cytotoxic effects on host cells. This work aims to generate a library that contains each of Phayonce’s 77 genes in an inducible expression vector. Individually cloned genes can then be introduced into M. smegmatis host cells for subsequent analysis of gene function. Each Phayonce gene was amplified by PCR using gene specific primers. Amplified genes were then assembled into the pExTra plasmid, which contains a tetracycline inducible promoter to drive Phayonce gene expression. After assembly and transformation into E.coli, colonies containing putative plasmids were analyzed by PCR to verify the insert. All 77 genes of Phayonce’s genome were successfully cloned into pExTra and subsequent sequencing established they are error-free. Generating a complete gene library of the bacteriophage Phayonce has allowed for all 77 expression vectors to be used in phenotypic analysis of toxicity of each Phayonce gene. Toxicity of Phayonce genes on host M. smegmatis may have medically significant implications on bacteriophage therapies for bacterial species that have shown to be resistant to treatment.