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| Predicate | Object |
|---|---|
| rdf:type | |
| lifeskim:mentions | |
| pubmed:issue |
3
|
| pubmed:dateCreated |
1994-6-15
|
| pubmed:abstractText |
Plasmid pSI30 was constructed to increase the sensitivity of detection of a genetically engineered micro-organism (GEM) and its recombinant DNA in environmental samples. This 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. This catabolic phenotype was exploited using enrichment procedures to detect RC-4(pSI30) cells, free-living 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. Using the eukaryotic DNA sequence as a probe, no transfer of the plasmid to indigenous bacteria was detected. Persistence of RC-4(pSI30) and its ability to multiply upon addition of 3-chlorobenzoate were demonstrated 78 days after its addition to natural freshwater. In flow-through microcosms RC-4(pSI30), undetectable as free-living 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.
|
| pubmed:language |
eng
|
| pubmed:journal | |
| pubmed:citationSubset |
IM
|
| pubmed:chemical | |
| pubmed:status |
MEDLINE
|
| pubmed:month |
Oct
|
| pubmed:issn |
0962-1083
|
| pubmed:author | |
| pubmed:issnType |
Print
|
| pubmed:volume |
1
|
| pubmed:owner |
NLM
|
| pubmed:authorsComplete |
Y
|
| pubmed:pagination |
137-43
|
| pubmed:dateRevised |
2006-11-15
|
| pubmed:meshHeading |
pubmed-meshheading:1344990-Biodegradation, Environmental,
pubmed-meshheading:1344990-Chlorobenzoates,
pubmed-meshheading:1344990-DNA, Recombinant,
pubmed-meshheading:1344990-DNA Probes,
pubmed-meshheading:1344990-Drug Resistance, Microbial,
pubmed-meshheading:1344990-Ecosystem,
pubmed-meshheading:1344990-Fresh Water,
pubmed-meshheading:1344990-Genetic Engineering,
pubmed-meshheading:1344990-Plasmids,
pubmed-meshheading:1344990-Pseudomonas putida,
pubmed-meshheading:1344990-Water Microbiology
|
| pubmed:year |
1992
|
| pubmed:articleTitle |
Use of a novel plasmid to monitor the fate of a genetically engineered Pseudomonas putida strain.
|
| pubmed:affiliation |
US Environmental Protection Agency, Environmental Research Laboratory, Gulf Breeze, FL 32561.
|
| pubmed:publicationType |
Journal Article
|