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Volume 148, Issue 11

Analysis of 16S libraries of mouse gastrointestinal microflora reveals a large new group of mouse intestinal bacteria

bThe GenBank accession numbers for the clone sequences reported in this paper can be found in Table 1 T1 ; the accession number for isolate MIB-CB3 is AJ418059.

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Abstract

Total genomic DNA from samples of intact mouse small intestine, large intestine, caecum and faeces was used as template for PCR amplification of 16S rRNA gene sequences with conserved bacterial primers. Phylogenetic analysis of the amplification products revealed 40 unique 16S rDNA sequences. Of these sequences, 25% (10/40) corresponded to described intestinal organisms of the mouse, including spp., spp., segmented filamentous bacteria and members of the altered Schaedler flora (ASF360, ASF361, ASF502 and ASF519); 75% (30/40) represented novel sequences. A large number (11/40) of the novel sequences revealed a new operational taxonomic unit (OTU) belonging to the –– phylum, which the authors named ‘mouse intestinal bacteria’. 16S rRNA probes were developed for this new OTU. Upon analysis of the novel sequences, eight were found to cluster within the – group and three clustered within the group. One of the novel sequences was distantly related to and one was distantly related to . Oligonucleotide probes specific for the 16S rRNA of these novel clones were generated. Using a combination of four previously described and four newly designed probes, approximately 80% of bacteria recovered from the murine large intestine and 71% of bacteria recovered from the murine caecum could be identified by fluorescence hybridization (FISH).

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Keyword(s): Bacteroides , commensal bacteria , DAPI, 4′,6-diamidino-2-phenylindole , FISH, fluorescence in situ hybridization , intestinal colonization , MIB, ‘mouse intestinal bacteria’ , OTU, operational taxonomic unit and SFB, segmented filamentous bacteria
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2002-11-01
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Analysis of 16S libraries of mouse gastrointestinal microflora reveals a large new group of mouse intestinal bacteriabbThe GenBank accession numbers for the clone sequences reported in this paper can be found in Table 1T1; the accession number for isolate MIB-CB3 is AJ418059.
Microbiology 148, 3651 (2002); https://doi.org/10.1099/00221287-148-11-3651
/content/journal/micro/10.1099/00221287-148-11-3651
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