Gene Transfer Agents: Prevalence, Biology and Impact on Bacterial Genetic Diversity. (360G-Wellcome-109363_Z_15_Z)

Gene transfer agents (GTAs) have been discovered in diverse prokaryotes, however, the prevalence of functional GTAs in pathogenic species is unknown. I have preliminary evidence that GTAs are present in numerous pathogens and experimental evidence of GTAs in Burkholderia. This project aims to understand the role of GTAs in the evolution of bacterial pathogens. Specifically: 1) To gain an understanding of the prevalence of GTAs in pathogenic bacteria by screening a bank of pathogenic bacteria for GTA production (e.g. Pseudomonas, Burkholderia, Neisseria, Clostridia). Discovery of a broader database of functional GTAs will allow robust identification of GTAs by bioinformatics. 2) To assess the impact of GTAs on the transfer of antibiotic and virulence genes between pathogenic bacteria using microcosms that approximate environmental and clinical conditions, and by exposing the producer bacteria to environmentally relevant stresses such as antibiotics or competitor species. 3) To understand the regulation of GTA induction using a bespoke flow cytometry cell sorting technique to separate GTA-producer cells from non-producers for analysis with transcriptomic and epigenetic analyses. 4) To determine the mechanism of random DNA packaging using in vitro and in vivo packaging assays based on established techniques from enterobacteria phage T4.