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The gene encoding the green fluorescent protein (GFP) from the jellyfish Aequorea victoria was assembled into an expression cassette for bacteria by fusing it to the T7 gene10 ribosome binding site and the strong, constitutive promoter PpsbA from Amaranthus hybridus. By using Tn5-based transposon delivery systems, Pseudomonas Jluorescens bacteria were chromosomally tagged with gfi. We demonstrate that expression of a single copy &p gene is sufficient to permit the visuaiization of bacteria by epifluorescence and laser confocal microscopy and detection by flow cytometry. The green fluorescent phenotype was detectable in all growth phases even under nutrient-limited conditions.
Tombolini et al. (Tue,) studied this question.