FROM GENE TO PROTEIN – CLONNING, EXPRESSION AND PUFICATION OF A P450 CYTOCHROM FROM Campylobacter jejuni
Keywords:
cytochrome P450, cloning, expression, purificationAbstract
Recently, the complete genome sequence of Campylobacter jejuni NCTC 11168 was
published revealing the presence of only one open reading frame (Cj1411c)
encoding for a cytochrome P450, in contrast to 20 found in M. tuberculosis. The
gene Cj1411c encodes for a soluble 52.6 kDa protein with a predicted isoelectric
point of 9.3. The P450 gene is part of reading frame which hosts genes involved in
the synthesis 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-Fischer syndromes. The structure
of the lipoolygosaccharides and capsule polysaccharide was published last year
revealing that the strain possessed a type II/III capsule locus found in other
microorganisms such Nisseria meningitidis. This project focuses on the cloning and
characterisation of the only P450 enzyme of the human pathogen Campylobacter
jejuni NCTC 11168. We aim to understand the metabolic role of this P450
cytochrome in order to elucidate its possible use as a new target for drug design. To
achieve this aim we have cloned, expressed and purify the product of P450 coding
gene.
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