FROM GENE TO PROTEIN – CLONNING, EXPRESSION AND PUFICATION OF A P450 CYTOCHROM FROM CAMPYLOBACTER JEJUNI

Recently, the complete genome sequence of Campylobacter jejuni NCTC 11168 was published revealingthe presence of only one open reading frame (Cj1411c) encoding for a cytochrome P450, in contrast to 20found in M. tuberculosis. The gene Cj1411c encodes for a soluble 52.6 kDa protein with a predictedisoelectric point of 9.3. The P450 gene is part of reading frame which hosts genes involved in thesynthesis of cell surface components (capsula). Campylobacter capsule are important in adherence,invasion and colonisation of host cells and for maintenance of cell surface charge and serum resistance.These capsule are thought to cause autoimmunity leading to Guillan-Barre and Miller-Fischersyndromes. The structure of the lipoolygosaccharides and capsule polysaccharide was published last yearrevealing that the strain possessed a type II/III capsule locus found in other microorganisms suchNisseria meningitidis. This project focuses on the cloning and characterisation of the only P450 enzymeof the human pathogen Campylobacter jejuni NCTC 11168. We aim to understand the metabolic role ofthis P450 cytochrome in order to elucidate its possible use as a new target for drug design. To achievethis aim we have cloned, expressed and purify the product of P450 coding gene.