||Use of a Novel Plasmid to Monitor the Fate of a Genetically Engineered 'Pseudomonas putida' Strain.
Genthner, F. J. ;
Campbell, R. P. ;
Pritchard, P. H. ;
||Environmental Research Lab., Gulf Breeze, FL. ;Technical Resources, Inc., Gulf Breeze, FL.
Pseudomonas putida ;
Genetic engineering ;
Antibiotic resistance ;
Deoxyribonucleic acids ;
Nucleic acid hybridization ;
Aquatic microbiology ;
||Most EPA libraries have a fiche copy filed under the call number shown. Check with individual libraries about paper copy.
||Plasmid pSI30 was constructed to increase the sensitivity of detection of a genetically engineered microorganism (GEM) and its recombinant DNA in environmental samples. The broad host-range, mobilizable plasmid contained chlorocatechol (clc) degradative genes, antibiotic resistance genes (ampicillin and kanamycin), and a fragment of eukaryotic DNA. The clc genes encode enzymes that convert 3-chlorocatechol to maleylacetic acid permitting the host, Pseudomonas putida RC-4, to grow on 3-chlorobenzoate. The catabolic phenotype was exploited using enrichment procedures to detect RC-4(pSI30) cells, freeliving in the water column or when irreversibly bound to surfaces. The eukaryotic DNA sequence provided a unique target allowing positive identification by DNA:DNA hybridization. In flow-through microcosms, RC-4(pSI30), undetectable as freeliving cells, was found by enrichment as irreversibly bound sessile forms. These experiments revealed the stability of pSI30 and its utility in a combination detection system for tracking the survival of a GEM and its DNA in environmental samples.
||Pub. in Molecular Ecology 1, p137-143 1992. See also PB91-206870. Prepared in cooperation with Technical Resources, Inc., Gulf Breeze, FL.
|NTIS Title Notes
||Reprint: Use of a Novel Plasmid to Monitor the Fate of a Genetically Engineered 'Pseudomonas putida' Strain.
||PC A02/MF A01