Application of a suite of 16S rRNA-specific oligonucleotide probes designed to investigate bacteria of the phylum cytophaga-flavobacter-bacteroides in the natural environment
Application of a suite of 16S rRNA-specific oligonucleotide probes designed to investigate bacteria of the phylum cytophaga-flavobacter-bacteroides in the natural environment
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DOI：
10.1099/13500872-142-5-1097
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发表时间：
1996-05-01
期刊：
影响因子：
2.8
通讯作者：
Schleifer, KH
中科院分区：
文献类型：
--
作者：
Manz, W;Amann, R;Schleifer, KH
我们设计了一组四个16 S rRNA靶向的寡核苷酸探针特异性噬细胞-黄杆菌-类杆菌（CFB）的门细菌。探针CF 319 a和CF 319 b靶向黄杆菌-噬细胞菌群和卟啉单胞菌属的成员，而探针BAC 303具有类杆菌群内的普雷沃氏菌属和拟杆菌属的靶区域特征。FFE 8b探针是为了与锈赤黄杆菌进行种特异性杂交而开发的。通过计算机辅助序列分析设计所有探针，并与所有目前可获得的16 S和23 S rRNA序列进行比较。通过全细胞和非放射性斑点印迹杂交针对CFB门和细菌的其他主要谱系的参考菌株进一步评估寡核苷酸。新开发的探针与其他高阶探针一起用于分析复杂环境中的结构和群落组成。在活性污泥样品中，通过原位杂交揭示了黄杆菌-噬纤维菌群的成员作为污泥絮体的重要组成部分和丝状菌的特征定殖者，通过应用荧光探针BAC 303，可以可视化拟杆菌属和普雷沃氏菌属的成员，而无需预先培养作为人类粪便微生物菌群的重要组成部分。
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.