Monitoring of Pseudomonas pseudoalcaligenes KF707 Labeled with gfp As a Reporter Gene

초록

The development of sensitive methods for observing individual bacterial cells in a population in experimental models and environments is very important. Use of the green fluorescent protein (GFP) from the jellyfish Aequorea victoria is a powerful method for nondestructive in situ monitoring, since expression of green fluorescence does not require any additional substrate. In this study, we have genetically marked Pseudomonas pseudoalcaligenes that is a PCB (polychlorinated biphenyl)-degrading soil Gram-negative bacterium with gfp. Using electroporation, we transfered gfp gene containing plasmid pJBA36 (mini-Tn5::gfp) to P. pseudoalcaligenes KF707. Expression of GFP did not alter the growth in the presence of PCB, and was detectable by epifluorescence microscopy, which would allow future monitoring of GFP+ P. pseudoalcaligenes strain in ecosystem.