Record Display for the EPA National Library Catalog


Main Title Use of a Novel Plasmid to Monitor the Fate of a Genetically Engineered 'Pseudomonas putida' Strain.
Author Genthner, F. J. ; Campbell, R. P. ; Pritchard, P. H. ;
CORP Author Environmental Research Lab., Gulf Breeze, FL. ;Technical Resources, Inc., Gulf Breeze, FL.
Publisher c1992
Year Published 1992
Report Number EPA/600/J-93/066;
Stock Number PB93-169001
Additional Subjects Pseudomonas putida ; Genetic engineering ; Antibiotic resistance ; Plasmids ; Deoxyribonucleic acids ; Nucleic acid hybridization ; Aquatic microbiology ; Biodeterioration ; Phenotype ; Growth ; Reprints ;
Library Call Number Additional Info Location Last
NTIS  PB93-169001 Some EPA libraries have a fiche copy filed under the call number shown. 07/26/2022
Collation 9p
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.