Presentation Type
Poster
Full Name of Faculty Mentor
Michael Pierce, Biology
Major
Biology
Presentation Abstract
Bacteriophage or phage are a diverse class of viruses that infect and reproduce in bacterial cells. Their diverse genomes represent an immense source of novel protein functions and a deeper understanding of phage genes will contribute to emerging treatments for antibiotic resistant bacterial infections. The goal of the research described here is to isolate and study individual genes from the bacteriophage Phayonce. Gene specific primers were used to PCR amplify Phayonce genes 12 and 77 (Phayonce 12 and Phayonce 77). The PCR products were then ligated into a plasmid vector by isothermal assembly. Following confirmation of the cloned gene by colony PCR, plasmids were transformed by electroporation into the Phayonce host bacterium, Mycobacterium smegmatis. To determine if the isolated phage genes interfered with cellular function, Phayonce genes were overexpressed from an inducible promoter in the host M. Smegmatis. Three isolated colonies overexpressing Phayonce 12 or Phayonce 77 were tested and comparted to toxic and non-toxic control strains of M. Smegmatis. Phayonce 12 overexpression did not result in a cytotoxic phenotype while the overexpression of Phayonce 77 did produce a cytotoxic phenotype. Future experiments will attempt to identify specific host proteins that interact with the proteins encoded by Phayonce genes 12 and 77.
Location
Poster Session 2
Start Date
13-4-2022 4:30 PM
End Date
13-4-2022 6:30 PM
Disciplines
Biology
Recommended Citation
Wilson, Amber, "Cloning and Overexpression of Phayonce Genes 12 and 77 in M. Smegmatis" (2022). Undergraduate Research Competition. 85.
https://digitalcommons.coastal.edu/ugrc/2022/fullconference/85
Included in
Cloning and Overexpression of Phayonce Genes 12 and 77 in M. Smegmatis
Poster Session 2
Bacteriophage or phage are a diverse class of viruses that infect and reproduce in bacterial cells. Their diverse genomes represent an immense source of novel protein functions and a deeper understanding of phage genes will contribute to emerging treatments for antibiotic resistant bacterial infections. The goal of the research described here is to isolate and study individual genes from the bacteriophage Phayonce. Gene specific primers were used to PCR amplify Phayonce genes 12 and 77 (Phayonce 12 and Phayonce 77). The PCR products were then ligated into a plasmid vector by isothermal assembly. Following confirmation of the cloned gene by colony PCR, plasmids were transformed by electroporation into the Phayonce host bacterium, Mycobacterium smegmatis. To determine if the isolated phage genes interfered with cellular function, Phayonce genes were overexpressed from an inducible promoter in the host M. Smegmatis. Three isolated colonies overexpressing Phayonce 12 or Phayonce 77 were tested and comparted to toxic and non-toxic control strains of M. Smegmatis. Phayonce 12 overexpression did not result in a cytotoxic phenotype while the overexpression of Phayonce 77 did produce a cytotoxic phenotype. Future experiments will attempt to identify specific host proteins that interact with the proteins encoded by Phayonce genes 12 and 77.