Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
5
pubmed:dateCreated
2001-6-20
pubmed:databankReference
pubmed:abstractText
Alpha subunit genes of initial polyaromatic hydrocarbon (PAH) dioxygenases were used as targets for the PCR detection of PAH-degrading strains of the genera Pseudomonas, Comamonas and Rhodococcus which were obtained from activated sludge or soil samples. Sequence analysis of PCR products from several Pseudomonas strains showed that alpha subunits (nahAc allele) of this genus are highly conserved. PCR primers for the specific detection of alpha subunit genes of initial PAH dioxygenases from Pseudomonas strains were not suitable for detecting the corresponding genes from the genera Comamonas and Rhodococcus. Southern analysis using a heterologous gene probe derived from the P. putida OUS82 PAH dioxygenase alpha subunit identified segments of the PAH-degradation gene cluster from C. testosteroni strain H. Parts of this gene cluster containing three subunits of the initial PAH dioxygenase were isolated. These three subunits [ferredoxin (pahAb), alpha (pahAc) and beta (pahAd) subunit] were amplified by PCR as one fragment and expressed in Escherichia coli DH5alpha, resulting in an active initial dioxygenase with the ability to transform indole and phenanthrene. The DNA sequence alignment of alpha subunits from C. testosteroni H and various PAH-degrading bacteria permitted the design of new primers and oligonucleotide probes which are useful for the detection of the initial PAH dioxygenases from strains of Pseudomonas, Comamonas and Rhodococcus.
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
May
pubmed:issn
0175-7598
pubmed:author
pubmed:issnType
Print
pubmed:volume
55
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
609-18
pubmed:dateRevised
2008-11-21
pubmed:meshHeading
pubmed-meshheading:11414329-Bacterial Proteins, pubmed-meshheading:11414329-Base Sequence, pubmed-meshheading:11414329-Biodegradation, Environmental, pubmed-meshheading:11414329-Comamonas, pubmed-meshheading:11414329-DNA Primers, pubmed-meshheading:11414329-Dioxygenases, pubmed-meshheading:11414329-Escherichia coli, pubmed-meshheading:11414329-Genes, Bacterial, pubmed-meshheading:11414329-Genetic Variation, pubmed-meshheading:11414329-Molecular Sequence Data, pubmed-meshheading:11414329-Multigene Family, pubmed-meshheading:11414329-Oxygenases, pubmed-meshheading:11414329-Phylogeny, pubmed-meshheading:11414329-Polycyclic Hydrocarbons, Aromatic, pubmed-meshheading:11414329-Polymerase Chain Reaction, pubmed-meshheading:11414329-Protein Subunits, pubmed-meshheading:11414329-Pseudomonas, pubmed-meshheading:11414329-Rhodococcus
pubmed:year
2001
pubmed:articleTitle
Insights into the genetic diversity of initial dioxygenases from PAH-degrading bacteria.
pubmed:affiliation
Technische Universität Berlin, Institut für Biotechnologie, Mikrobiologie und Genetik, Germany.
pubmed:publicationType
Journal Article, Research Support, Non-U.S. Gov't