Molecular analysis and development of 16S rRNA oligonucleotide probes to characterize a diclofop-methyl-degrading biofilm consortium
作者:
Louise Laramée,
John R Lawrence,
Charles W Greer,
期刊:
Canadian Journal of Microbiology
(NRC Available online 2000)
卷期:
Volume 46,
issue 2
页码: 133-142
ISSN:0008-4166
年代: 2000
DOI:10.1139/w99-129
出版商: NRC Research Press
数据来源: NRC
摘要:
Genomic DNA from nine individual bacteria, isolated from a diclofop-methyl-degrading biofilm consortium, was extracted for genetic characterization. The degradation of diclofop-methyl produces metabolites that are known intermediates or substrates for bacteria that degrade a variety of chlorinated aromatic compounds. Accordingly, oligonucleotide primers were designed from specific catabolic genes for chlorinated organic degradation pathways, and tested by PCR to determine if these genes are involved in diclofop-methyl degradation. DNA homology between the PCR products and the known catabolic genes investigated by Southern hybridization analysis and by sequencing, suggested that novel catabolic genes are functioning in the isolates. Specific fluorescent oligonucleotides were designed for two of the isolates, following 16S rDNA sequencing and identification of each of the isolates. These probes were successfully used for fluorescent in situ hybridization (FISH) studies of the two isolates in the biofilm consortium.Key words: consortium, catabolic gene, diclofop-methyl, 16S rDNA, FISH, SCLM.
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