Interactions between the gut microbiome and pathogenic bacteria can influence disease outcomes. Vibrio cholerae, the causative agent of cholera, infects the small intestine. During infection, V. cholerae can interact with Streptococcus, a component of the gut microbiota elevated in dysbiosis, a state of microbial imbalance that sensitizes patients to cholera infection. To determine local cell density through quorum sensing, Streptococcus produces and detects a short hydrophobic peptide (SHP). SHP also increases the motility of V. cholerae by interacting with the flagellar motor proteins PomA and PomB, subsequently increasing sodium ion flow across the cell envelope, which powers flagellar rotation. With motility as a virulence factor for V. cholerae, an increase in Streptococcus density in dysbiosis leads to worse patient outcomes. In this project, we aimed to identify mutations in the pomA gene that reduce the responsiveness of V. cholerae to SHP to further understand the SHP-PomA interaction. Using error-prone PCR, we induced random mutations in the pomA gene and screened more than 500 mutants for unresponsiveness to SHP using high-throughput live cell microscopy. Ultimately, we identified two mutant strains unresponsive to the peptide and sequenced them to identify the mutations responsible for this phenotype. Future experiments will involve coculturing Streptococcus and identified mutant V. cholerae strains in mouse models to examine whether these mutations reduce the virulence of cholera in vivo. Overall, investigating these mutations enhances understanding of the interaction between V. cholerae and Streptococcus and informs potential novel treatment methods that consider how microbiota affects cholera outcomes.