Initiation and impact of conjugative transfer by antibiotic resistant haemophilus element. (360G-Wellcome-077183_Z_05_Z)

The spread of antibiotic resistance is a major threat to the continued successof medicine. By studying the evolution and properties of antibiotic resistancein Haemophilus influenzae, which arises from the successful spread of a singlefamily of conjugating and integrating elements (ICEs) world wide, an explanatory model is possible. Furthermore, these ICEs are evolutionarily related to a number of well known pathogenicity islands. This provides a research opportunity for testing widely held hypotheses about the emergence and spread of antibiotic resistance or genomic islands. The proposed training fellowship aims to contribute to this by investigating first the initiation ofconjugation by ICEs and second provide proof in principle that these ICEs may contribute to the diversity in H. influenzae by the following approaches:1. The oriT of ICEHin1056 will be identified and characterised. The role of traI and its regulation will be studied using a combination of cloning, mutagenesisand transcriptional analysis using microarrays. 2. The role of ICEs in the genetic diversification of H. influenzae will be studied by determining sequence variation in divergent sequences close to the putative oriT. The ability of ICEs to mobilise whole chromosomes in an Hfr-like mechanism will also be investigated.