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We have developed two sets ofCampylobacter shuttle vectors containing either thegfp (green fluorescent protein),yfp (yellow fluorescent protein), orcfp (cyan fluorescent protein) reporter gene. In one set, the reporter gene is fused to a consensusCampylobacter promoter sequence (Pc ). The other set contains a pUC18 multicloning site upstream of the reporter gene, allowing the construction of transcriptional fusions using known promoters or random genomic fragments.C. jejuni cells transformed with the Pc fusion plasmids are strongly fluorescent and easily visualized on chicken skin, on plant tissue, and within infected Caco-2 cells. In eachC. jejuni strain tested, these plasmids were maintained over several passages in the absence of antibiotic selection. Also, in manyC. jejuni strains, &gt;91% of the cells transformed with the Pc fusion plasmids remained fluorescent after several days. Experiments with yellow fluorescent and cyan fluorescentC. jejuni transformants suggest that aggregates containing two or more strains ofC. jejuni may be present in an enrichment broth culture. Colonies arising from these aggregates would be heterologous in nature; therefore, isolation of a pure culture ofC. jejuni , by selecting single colonies, from an environmental sample may not always yield a single strain.

Detection on Surfaces and in Caco-2 Cells of Campylobacter jejuni Cells Transformed with New gfp, yfp , and cfp Marker Plasmids