PHYSICAL AND GENETIC MAP OF THE GENOME OF CAMPYLOBACTER-UPSALIENSIS

A physical map of Campylobacter upsaliensis ATCC 43954 was constructed from DNA fragments generated by Sa/I (5' G/TCGAC), NarI (5' GG/CGCC) and BssHII (5' G/CGCGC) restriction digests separated using pulsed-field gel electrophoresis. The size of the C. upsaliensis genome was approximately 2000 kb, providing evidence of the largest Campylobacter genome sized to date. Twenty-one fragments created from these restriction digests were assembled into a physical map using a combination of complementary methods including cross-Southern hybridization, hybridiration fingerprint analysis and hybridization with homologous and heterologous (from Campylobacter ieiunr3 gene probes. The position of ten genetic loci, including that of the iron-uptake regulatory (fur) gene, were localized to the physical map. A genomic library of C. upsaliensis ATCC 43954 was constructed in lambda Gem-11 vector. Fifty thousand recombinants with an average sire of 16 kb represent a library about 200 times the size of the genome. Using C. jejuni DNA probes, clones representing C. upsaliensis flaA, fur and ftsZ genes were isolated and localized to the physical map.