Investigating the potential role of Azabicyclene in the Rhl Quorum Sensing Pathway of Pseudomonas aeruginosa

Pseudomonas aeruginosa is a gram-negative bacterial pathogen responsible for many antibiotic-resistant infections. Through a process called quorum sensing, P. aeruginosa is able to communicate with other cells causing the infection to persist. The production of the molecule azabicyclene is regulated through one of the key quorum sensing pathways. However, the physiological role Azabicyclene plays in P. aeruginosa remains unknown. Here we report the effects of Azabicyclene on the Rhl quorum sensing pathway. By deleting key genes to both the Rhl quorum sensing pathway and the production of azabicyclene, a multiple strains were engineered to examine the potential effect of azabicyclene. We experienced difficulties attempting to delete key genes from both the aze operon and Rhl QS pathway. Despite the setbacks, four unique strains of P. aeruginosa were created. Supernatants from these engineered strains were applied to luciferase reporters to determine the differences in gene expression. Determining the role of azabicyclene in P. aeruginosa allows for continuation to develop targeted antibiotics and increases knowledge of quorum sensing.