Development of molecular tools for characterization and exploitation of a library of bacteriophages infecting Pseudomonas aeruginosa.

Bacterial viruses (bacteriophages) are the most abundant and genetically diverse biological entities on this planet. Billion years of coevolution with their bacterial host organisms enabled the development of various phage-encoded mechanisms to shut down the host metabolism and direct it towards phage production. My doctoral research focused on the genetic diversity of bacteriophages infecting Pseudomonas aeruginosa, an important multi-drug resistant opportunistic pathogen. The described research project aims initially at the completion of the genome and proteome analysis of two Pseudomonas phages, LIT1 and LUZ7. This will be followed by the start-up of three parallel research lines to identify and characterize phage-encoded antibacterial proteins. In collaboration with Dr. Bart Landuyt (Afdeling Dierenfysiologie en Neurobiologie, K.U.Leuven), all peptides of a phage-infected cell will be screened using the 'Peptidomics' approach, leading to the experimental identification of (un)annotated phage peptides. Secondly, the suitability of 'Tandem Affinity Purification' to identify interactions between phage proteins and different subunits of the bacterial RNA polymerase will be evaluated. Finally, the recombineering system, which catalyses in vivo homologous recombination between linear DNA substrates and their counterpart in the phage genome, will be optimized in Pseudomonas aeruginosa to create specific virulent phage mutants. These three research lines will be integrated into each other in a later stage of research, enabling future study of phage-encoded antibacterials.

Keywords:Pseudomonas, Genome analysis, Tandem Affinity Purification, Bacteriophage, Recombineering, Peptidomics

Disciplines:Other engineering and technology