Record Display for the EPA National Library Catalog
RECORD NUMBER: 525 OF 5694
|OLS Field Name||OLS Field Data|
|Main Title||Assay for Detection and Enumeration of Genetically Engineered Microorganisms Which Is Based on the Activity of a Deregulated 2,4-Dichlorophenoxyacetate Monooxygenase.|
|Author||King, R. J. ; Short, K. A. ; Seidler, R. J. ;|
|CORP Author||Corvallis Environmental Research Lab., OR. ;NSI Technology Services Corp., Corvallis, OR. Environmental Research Lab.|
|Additional Subjects||Assaying ; Genetics ; Microorganisms ; Chemical analysis ; Biotechnology ; Phenol ; Cells(Biology) ; Cell cultures ; Oxygenases ; Chromatographic analysis ; Reprints ; Oxygenase/2-4-Dichlorophenoxyacetate ; Genetic engineering ; Pseudomonas aeruginosa ; Pseudomonas putida|
An assay system was developed for the enumeration of genetically engineered microorganisms expressing a deregulated 2,4-dichlorophenoxyacetate (TFD) monooxygenase, which coverts phenoxyacetate (PAA) to phenol. In PAA-amended cultures of Pseudomonas aeruginosa PAO1C(pRO103) and Pseudomonas putida PPO301(pRO103), strains which express a deregulated TFD monoxygenase, phenol production was proprtional to cell number. Phenol was reacted, under specific conditions, with a 4-aminoantipyrine dye to form an intensely colored dye-phenol complex (AAPPC), which when measured spectrophotmetrically could detect as few as 10(3) cells per ml. The assay was corrobrated by monitoring the disappearnace of PAA and the accumulation of phenol by high-performance liquid chromatography and gas chromatography.