Amplification of putative chlorocatechol dioxygenase gene fragments from alpha- and beta-Proteobacteria.
Redundant primers were designed for the PCR amplification of DNA from chlorocatechol dioxygenase genes. These primers were used successfully to amplify 270- to 279-bp fragments from a variety of 2,4-dichlorophenoxyacetate- and cholorobenzoate-degrading strains, including species of Sphingomonas. Three groups of closely related sequences were amplified: one from chlorobenzoate degraders that was 86% similar to the amino acid sequence of the protein coded by the tfdC gene of Ralstonia eutropha JMP134 (pJP4), a second from Sphingomonas strains that was 70% similar to this amino acid sequence, and a third from diverse 2,4-D degraders that showed only 53% similarity to the product coded by tfdC from pJP4 but 88-100% similarity to the product of the tfdC gene of the plasmid pEST4011 from a Pseudomonas putida strain. The primers should be useful in further study of this gene and in tracking a variety of degraders of chloroaromatic compounds in natural systems.
Leander
M
M
Division of Physical Sciences, University of Toronto, ON, Canada.
Vallaeys
T
T
Fulthorpe
R
R
D015084:2,4-Dichlorophenoxyacetic Acid / Q000378:metabolism*; D000595:Amino Acid Sequence; D001673:Biodegradation, Environmental; D002723:Chlorobenzoates / Q000378:metabolism*; D017931:DNA Primers; D049308:Dioxygenases; D005798:Genes, Bacterial; D006090:Gram-Negative Bacteria / Q000201:enzymology* / Q000235:genetics*; D008969:Molecular Sequence Data; D010105:Oxygenases / Q000235:genetics* / Q000378:metabolism; D016133:Polymerase Chain Reaction; D017422:Sequence Analysis, DNA; D012988:Soil Microbiology; D014871:Water Microbiology
D002723:Chlorobenzoates; D017931:DNA Primers; D015084:2,4-Dichlorophenoxyacetic Acid; D010105:Oxygenases; D049308:Dioxygenases; C053101:catechol oxygenase II