f In situ probing of Gram-positive bacteria with high DNA G + C content using 23S rRNA-targeted oligonucleotides
- Authors: Carsten Roller, Michael Wagner, Rudolf Amann*, Wolfgang Ludwig, Karl-Heinz Schleifer
- *Author for correspondence: Rudolf Amann. Tel: +49 89 2105 2373. Fax: +49 89 2105 2360.
- Microbiology, October 1994 140: 2849-2858, doi: 10.1099/00221287-140-10-2849
- Subject: Environmental Microbiology
- Published Online:
23S-rRNA-targeted oligonucleotide probes were designed for the phylogenetic group ‘Gram-positive bacteria with high G + C content of DNA’ (GPBHGC). A sequence idiosyncrasy in two adjacent base pairs in the stem of helix 69 in domain IV of the 23S rRNA is present in all hitherto analysed strains of GPBHGC. An oligonucleotide probe targeted to this region hybridized only with strains of GPBHGC and was successfully used for in situ monitoring of these cells in activated sludge. Another unique feature of the 23S rRNA molecules of GPBHGC is a large insertion in domain III. Fluorescent oligonucleotides targeted to the highly variable regions of the rRNA within the insertions of Corynebacterium glutamicum DSM 20300T, Aureobacterium testaceum DSM 20166 and Brevibacterium sp. DSM 20165 hybridized specifically to their target strains, whereas probing with oligonucleotides complementary to the rRNA-coding strand of the 23S rDNA and to the spacer between 16S and 23S rRNA of C. glutamicum did not result in detectable fluorescence. This confirmed that the large 23S insertions are indeed present in 23S rRNAs of GPBHGC and provide potential target sites for highly specific nucleic acid probes.
- Keyword(s): 23S rRNA, Gram-positive bacteria, oligonucleotide probes, in situ hybridization, insertion
© Society for General Microbiology 1994 | Published by the Microbiology Society
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