Fluorescent oligonucleotide rDNA probes for specific detection of methane oxidising bacteria

Bourne, David G., Holmes, Andrew J., Iversen, Niels, and Murrell, J Colin (2000) Fluorescent oligonucleotide rDNA probes for specific detection of methane oxidising bacteria. FEMS Microbiology Ecology, 31 (1). pp. 29-38.

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Abstract

Oligonucleotide probes targeting the 16S rRNA of distinct phylogenetic groups of methanotrophs were designed for the in situ detection of these organisms. A probe, MG-64, detected specifically type I methanotrophs. while probes MA-221 and MA-621, detected type II methanotrophs in whole cell hybridisations. A probe Mc1029 was also designed which targeted only organisms from the Methylococcus genus after whole cell hybridisations. All probes were labelled with the fluorochrome Cy3 and optimum conditions for hybridisation were determined. Non-specific target sites of the type I (MG-64) and type IT (MA-621) probes to non-methanotrophic organisms are highlighted. The probes are however used in studying enrichment cultures and environments when selective pressure favours the growth of methanotrophs over other organisms. The application of these probes was demonstrated in the detection of type I methanotrophs with the MG-64 probe in an enrichment culture from an estuarine sample demonstrating methane oxidation. The detection of type I methanotrophs was confirmed by a 16S rDNA molecular analysis of the estuarine enrichment culture which demonstrated that the most abundant bacterial clone type in the 16S rDNA library was most closely related to Methylobacter sp. strain BB5.1, a type I methanotroph also isolated from an estuarine environment. (C) 2000 Federation of European Microbiological Societies. Published by Elsevier Science B.V. All rights reserved.

Item ID: 51160
Item Type: Article (Research - C1)
ISSN: 0168-6496
Keywords: methane oxidising bacteria, 16S rDNA probe, fluorescent in situ hybridisation
Funders: European Community RTD Biotechnology Program (BIO)
Projects and Grants: BIO 4 CT960419
Date Deposited: 11 Oct 2017 07:43
FoR Codes: 06 BIOLOGICAL SCIENCES > 0605 Microbiology > 060599 Microbiology not elsewhere classified @ 100%
SEO Codes: 96 ENVIRONMENT > 9699 Other Environment > 969999 Environment not elsewhere classified @ 100%
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