Characterization of a Bacteriophage Protein That Is Toxic to Mycobacterium Smegmatis

Overview

Antibiotic resistance is one of the most pressing public health concerns of our time. Currently, antibiotic resistance causes nearly 3 million infections and over 35,000 deaths in the U.S annually. Bacteriophages (phages) are viruses with a natural ability to infect and destroy bacteria. Due to this natural ability, they are being investigated as an alternative approach to battle antibiotic resistant bacteria. The goal of this project was to identify bacteriophage genes whose protein products are toxic to the host cell. This approach bypasses the need to use whole phage for therapeutic treatment. Cytotoxicity of bacteriophage gene products was determined by expressing cloned phage genes in Mycobacterium smegmatis and monitoring cell growth. Our analysis revealed that the expression of the bacteriophage MooMoo gene 86 protein product prevents cell growth. We then employed a 2-hybrid assay to identify the host protein(s) that interact with the MooMoo gene 86 protein product. Sequence analysis of host gene clones revealed multiple interacting partners. This pattern was also observed with our controls, suggesting that the conditions we employed lack sufficient specificity to identify true/known interactions. We are currently troubleshooting the 2 hybrid assay to improve its specificity.