Application of a suite of 16S rRNA-specific oligonucleotide probes designed to investigate bacteria of the phylum cytophaga-flavobacter-bacteroides in the natural environment

Penerapan serangkaian probe oligonukleotida spesifik-rRNA 16S yang dirancang untuk menginvestigasi bakteri dari filum Cytophaga-Flavobacter-Bacteroides di lingkungan alami

Clicks: 345
ID: 270213
1996
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Abstract
We designed a panel of four 16S rRNA-targeted oligonucleotide probes specific for bacteria of the phylum cytophaga-flavobacter-bacteroides (CFB). Probes CF319a and CF319b are targeted to members of the flavobacteria-cytophaga group and the genus Porphyromonas, whereas probe BAC303 has a target region characteristic for the genera Prevotella and Bacteroides within the bacteroides group. The probe FFE8b was developed for species-specific hybridizations with Flavobacterium ferrugineum. All probes were designed by computer-assisted sequence analysis and compared to all currently accessible 16S and 23S rRNA sequences. The oligonucleotides were further evaluated by whole-cell and non-radioactive dot-blot hybridization against reference strains of the CFB phylum and other major lineages of Bacteria. The newly developed probes were used together with other higher-order probes to analyse the structure and community composition in complex environments. In activated sludge samples, members of the flavobacteria-cytophaga group were revealed by in situ hybridization as important constituents of sludge flocs and characteristic colonizers of filamentous bacteria. By application of fluorescent probe BAC303, members of the genera Bacteroides and Prevotella could be visualized without prior cultivation as an important part of the human faecal microflora.
Kami merancang sebuah panel yang terdiri dari empat probe oligonukleotida bertarget rRNA 16S yang spesifik untuk bakteri dari filum cytophaga-flavobacter-bacteroides (CFB). Probe CF319a dan CF319b ditargetkan untuk anggota kelompok flavobacteria-cytophaga dan genus *Porphyromonas*, sedangkan probe BAC303 memiliki wilayah target yang khas untuk genus *Prevotella* dan *Bacteroides* di dalam kelompok bacteroides. Probe FFE8b dikembangkan untuk hibridisasi spesifik-spesies dengan *Flavobacterium ferrugineum*. Semua probe dirancang melalui analisis sekuens berbantuan komputer dan dibandingkan dengan seluruh sekuens rRNA 16S dan 23S yang saat ini dapat diakses. Oligonukleotida tersebut dievaluasi lebih lanjut melalui hibridisasi sel utuh dan *dot-blot* non-radioaktif terhadap galur referensi dari filum CFB dan garis keturunan utama Bakteri lainnya. Probe yang baru dikembangkan ini digunakan bersama dengan probe tingkat lebih tinggi lainnya untuk menganalisis struktur dan komposisi komunitas di lingkungan yang kompleks. Pada sampel lumpur aktif, anggota kelompok flavobacteria-cytophaga terungkap melalui hibridisasi *in situ* sebagai konstituen penting flok lumpur dan kolonisator karakteristik dari bakteri berfilamen. Melalui penerapan probe fluoresen BAC303, anggota genus *Bacteroides* dan *Prevotella* dapat divisualisasikan tanpa kultivasi sebelumnya sebagai bagian penting dari mikroflora fekal manusia.
Reference Key
manz1996microbiologyapplication Use this key to autocite in the manuscript while using SciMatic Manuscript Manager or Thesis Manager
Authors Werner Manz;Rudolf Amann;Wolfgang Ludwig;Marc Vancanneyt;Karl-Heinz Schleifer;Werner Manz;Rudolf Amann;Wolfgang Ludwig;Marc Vancanneyt;Karl-Heinz Schleifer;
Journal microbiology
Year 1996
DOI
doi:10.1099/13500872-142-5-1097
URL
http://dx.doi.org/10.1099/13500872-142-5-1097
Keywords
National Center for Biotechnology Information NCBI NLM MEDLINE RNA humans pubmed abstract nih national institutes of health national library of medicine research support non-u.s. gov't Comparative Study Ribosomal Oligonucleotide Probes* Base Sequence Molecular Sequence Data Phylogeny feces / microbiology 16s / genetics* bacteroides / genetics environmental microbiology* bacterial / genetics* pmid:8704951 doi:10.1099/13500872-142-5-1097 werner manz rudolf amann karl-heinz schleifer bacteroides / isolation & purification* cytophaga / genetics cytophaga / isolation & purification* flavobacterium / genetics flavobacterium / isolation & purification* gram-negative bacteria / classification*

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