Open AccessFlow cytometric and microscopic analysis of GFP‐tagged Pseudomonas fluorescens bacteria
Author(s) -
Tombolini Riccardo,
Unge Annika,
Davey Mary Ellen,
Bruijn Frans J,
Jansson Janet K
Publication year - 1997
Publication title -
fems microbiology ecology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.377
H-Index - 155
eISSN - 1574-6941
pISSN - 0168-6496
DOI - 10.1111/j.1574-6941.1997.tb00352.x
Subject(s) - aequorea victoria , biology , green fluorescent protein , pseudomonas fluorescens , bacteria , flow cytometry , microbiology and biotechnology , fluorescence microscope , gene , genetics , fluorescence , physics , quantum mechanics
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 P psbA from Amaranthus hybridus . By using Tn 5 ‐based transposon delivery systems, Pseudomonas fluorescens bacteria were chromosomally tagged with gfp . We demonstrate that expression of a single copy gfp gene is sufficient to permit the visualization 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.