Introduction of green fluorescent protein gene into phenol-degrading Alcaligenes faecalis cells and their monitoring in phenol-contaminated soil

Alcaligenesfaecalis (CCT 7145) was isolated from an Amazonian soil sample after an enrichment process to select for phenol-degrading microorganisms. The isolate was labeled with the green fluorescent protein (gfp) gene. The gfp-transformed cells were easily detected using a hand-held UV transillumin...

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Bibliographic Details
Published in:Applied microbiology and biotechnology 2001-07, Vol.56 (1-2), p.255-260
Main Authors: BASTOS, A. E. R, CASSIDY, M. B, TREVORS, J. T, LEE, H, ROSSI, A
Format: Article
Language:English
Subjects:
Alcaligenes
Alcaligenes - metabolism
Alcaligenes faecalis
Bacteria
Biodegradation of pollutants
Biological and medical sciences
Biotechnology
Environment and pollution
Environmental monitoring
Fundamental and applied biological sciences. Psychology
gfp gene
green fluorescent protein
Green Fluorescent Proteins
Industrial applications and implications. Economical aspects
Luminescent Proteins - genetics
Microorganisms
Mineralization
Phenol - metabolism
Phenols
Proteins
Soil contamination
Soil Microbiology
Soil Pollutants - metabolism
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Summary:Alcaligenesfaecalis (CCT 7145) was isolated from an Amazonian soil sample after an enrichment process to select for phenol-degrading microorganisms. The isolate was labeled with the green fluorescent protein (gfp) gene. The gfp-transformed cells were easily detected using a hand-held UV transilluminator and their taxonomy was confirmed by 16S rRNA sequencing. Polymerase chain reaction (PCR) and Southern blot analyses confirmed that the gfp gene was integrated into the chromosome. The addition of the gfp marker did not affect phenol degradation ability compared with the wild-type. Both, wild-type and gfp-marked A. faecalis cells encapsulated in alginate, tolerated 1,700 microg ml(-1) phenol in liquid medium compared with 1,100 microg ml(-1) phenol for free cells. 14C-Phenol mineralization in soil microcosms was also enhanced by inoculation with encapsulated cells. Survival of gfp-marked cells in phenol-contaminated soil over 22 days was determined from plate counts using an epifluorescence microscope.
ISSN:0175-7598
1432-0614
DOI:10.1007/s002530100614