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RECORD NUMBER: 8 OF 124

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.
Publisher cJun 91
Year Published 1991
Report Number EPA/600/J-94/469;
Stock Number PB95-137311
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
Holdings
Library Call Number Additional Info Location Last
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Status
NTIS  PB95-137311 Most EPA libraries have a fiche copy filed under the call number shown. Check with individual libraries about paper copy. NTIS 03/06/1995
Collation 5p
Abstract
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.