Gene Expression of Shival Phages Terminase Gene
Presentation Type
Poster
Faculty Advisor
Quinn Vega
Access Type
Event
Start Date
26-4-2024 12:45 PM
End Date
26-4-2024 1:44 PM
Description
For decades, bacteria have been evolving to become drug resistant. With increased antibiotic resistance, researchers are searching for new ways to fight off bacterial infections. My research is focused on studying bacteriophages, viruses that infect bacteria which may prove to be alternatives to the currently used antibiotics. One of these phages, Shival, has been isolated at Montclair State University and its genome has been sequenced. However, while this sequencing has led to the identification of proposed genes, the actual function of the protein products has not been confirmed. This goal of this project is to confirm the function of gene 2 of Shival, which is approximately 1800 base pairs long and is believed to code for the protein Terminase. In order to analyze protein function, the gene has been cloned through PCR amplification and the gene inserted into an expression vector. The plasmid was then transformed into bacteria and the plasmid isolated through a DNA mini-prep procedure. Restriction digestion and agarose gel analysis was used to confirm that the correct construct was made. This construct has also been partially sequenced to confirm isolation of the correct gene. After IPTG induction, a protein gel was used to confirm that the protein has been produced. Future studies will evaluate the protein’s enzymatic function after protein purification. Through this project, we hope to expand our understanding of phage gene function.
Gene Expression of Shival Phages Terminase Gene
For decades, bacteria have been evolving to become drug resistant. With increased antibiotic resistance, researchers are searching for new ways to fight off bacterial infections. My research is focused on studying bacteriophages, viruses that infect bacteria which may prove to be alternatives to the currently used antibiotics. One of these phages, Shival, has been isolated at Montclair State University and its genome has been sequenced. However, while this sequencing has led to the identification of proposed genes, the actual function of the protein products has not been confirmed. This goal of this project is to confirm the function of gene 2 of Shival, which is approximately 1800 base pairs long and is believed to code for the protein Terminase. In order to analyze protein function, the gene has been cloned through PCR amplification and the gene inserted into an expression vector. The plasmid was then transformed into bacteria and the plasmid isolated through a DNA mini-prep procedure. Restriction digestion and agarose gel analysis was used to confirm that the correct construct was made. This construct has also been partially sequenced to confirm isolation of the correct gene. After IPTG induction, a protein gel was used to confirm that the protein has been produced. Future studies will evaluate the protein’s enzymatic function after protein purification. Through this project, we hope to expand our understanding of phage gene function.