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Potential biological roles of the  Campylobacter jejuni  genes  cj0641, cj0774c  and  cj1663  were investigated. The proteins encoded by these genes showed sequence similarities to the phosphonate utilisation PhnH, K and L gene products of  Escherichia coli . The genes  cj0641, cj0774c  and  cj1663  were amplified from the pathogenic  C. jejuni  strain 81116, sequenced, and cloned into pGEM-T Easy vectors. Recombinant plasmids were used to disrupt each one of the genes by inserting a kanamycin resistance ( Km  R ) cassette employing site-directed mutagenesis or inverse PCR.  Campylobacter jejuni  81116 isogenic mutants were generated by integration of the mutated genes into the genome of the wild-type strain. The  C. jejuni  mutants grew on primary isolation plates, but they could not be purified by subsequent passages owing to cell death. The mutant  C. jejuni  strains survived and proliferated in co-cultures with wild-type bacteria or in media in which wild-type  C. jejuni  had been previously grown. PCR analyses of mixed wild-type/mutant cultures served to verify the presence of the mutated gene in the genome of a fraction of the total bacterial population. The data suggested that each mutation inactivated a gene essential for survival. Rates of phosphonate catabolism in lysates of  E. coli  strain DH5α were determined using proton nuclear magnetic resonance spectroscopy. Whole-cell lysates of the wild-type degraded phosphonoacetate, phenylphosphonate and aminomethylphosphonate. Significant differences in the rates of phosphonate degradation were observed between lysates of wild-type  E. coli , and of bacteria transformed with each one of the vectors carrying one of the  C. jejuni  genes, suggesting that these genes were involved in phosphonate catabolism.

Characterisation of Campylobacter jejuni genes potentially involved in phosphonate degradation