Rapid, Sensitive Bioluminescent Reporter Technology for Naphthalene Exposure and Biodegradation

A bioluminescent reporter plasmid for naphthalene catabolism (pUTK21) was developed by transposon (Tn4431) insertion of the lux gene cassette from Vibrio fischeri into a naphthalene catabolic plasmid in Pseudomonas fluorescens. The insertion site of the lux transposon was the nahG gene encoding for...

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Published inScience (American Association for the Advancement of Science) Vol. 249; no. 4970; pp. 778 - 781
Main Authors King, J. M. H., DiGrazia, P. M., Applegate, B., Burlage, R., Sanseverino, J., Dunbar, P., Larimer, F., Sayler, G. S.
Format Journal Article
LanguageEnglish
Published United States American Society for the Advancement of Science 17.08.1990
American Association for the Advancement of Science
The American Association for the Advancement of Science
Subjects
Acid soils
Bioassay
Biochemistry
Biodegradation
Biological assay
Bioluminescence
Biotechnology
Blood
Chemical detectors
Environmental cleanup
Genes
Genetics
Light
Naphthalene
Plasmids
Salicylates
Sensors
Slurries
Soil pollution
Soil water
Transposons
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Summary:A bioluminescent reporter plasmid for naphthalene catabolism (pUTK21) was developed by transposon (Tn4431) insertion of the lux gene cassette from Vibrio fischeri into a naphthalene catabolic plasmid in Pseudomonas fluorescens. The insertion site of the lux transposon was the nahG gene encoding for salicylate hydroxylase. Luciferase-mediated light production from P. fluorescens strains harboring this plasmid was induced on exposure to naphthalene or the regulatory inducer metabolite, salicylate. In continuous culture, light induction was rapid (15 minutes) and was highly responsive to dynamic changes in naphthalene exposure. Strains harboring pUTK21 were responsive to aromatic hydrocarbon contamination in Manufactured Gas Plant soils and produced sufficient light to serve as biosensors of naphthalene exposure and reporters of naphthalene biodegradative activity. The robust and sensitive nature of the bioluminescent reporter technology suggests that new sensing methods can be developed for on-line process monitoring and control in complex environmental matrices.
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ISSN:0036-8075
1095-9203
DOI:10.1126/science.249.4970.778