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Development of a signature probe targeting the 16S-23S rRNA internal transcribed spacer of a ruminal Ruminococcus flavefaciens isolate from reindeer.
MedLine Citation:
PMID:  21831789     Owner:  NLM     Status:  In-Data-Review    
Abstract/OtherAbstract:
The cellulolytic Ruminococcus flavefaciens has previously been introduced into the ruminant rumen to increase microbial degradation of plant cell wall carbohydrates. The functional effect of an introduced bacterium depends on its ability to establish in the digestive tract, and signature probes can be used as a tool to track and quantify introduced strains. The purpose of this current study was to develop an oligonucleotide signature probe targeting the 16S-23S rRNA internal transcribed spacer (ITS) of a putative probiotic cellulolytic isolate (R. flavefaciens strain 8/94-32) from the rumen of reindeer (Rangifer tarandus tarandus). The 16S-23S rRNA gene ITS of three Ruminococcus strains; R. flavefaciens strain 8/94-32, R. flavefaciens FD-1 and Ruminococcus albus Ra-8, was investigated. The ITS region has been reported to vary more between closely related bacteria compared to the widely used 16S rRNA gene, and a high degree of sequence polymorphism was indeed detected between the three Ruminococcus strains studied. Based on observed sequence differences, two oligonucloetide probes, ITSRumi1 and ITSRumi2, targeting the ITS region of the R. flavefaciens isolate 8/94-32 were developed. Probe specificity was evaluated in dot blot hybridisations with R. flavefaciens isolate 8/94-32 and four other Ruminococcus-strains tested. The probe ITSRumi1 gave positive signals for the R. flavefaciens isolate 8/94-32 only, while probe ITSRumi2 gave positive signals for R. flavefaciens isolate 8/94-32 as well as for R. albus Ra-8. The result of hybridisations with the probe ITSRumi1 indicates that the probe is specific for the R. flavefaciens strain 8/94-32 amongst the four Ruminococcus-strains tested, and is promising for further studies using it as a signature probe for tracking this strain when re-introduced to the reindeer rumen.
Authors:
K E Præsteng; R I Mackie; I K O Cann; S D Mathiesen; M A Sundset
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Publication Detail:
Type:  Journal Article    
Journal Detail:
Title:  Beneficial microbes     Volume:  2     ISSN:  1876-2891     ISO Abbreviation:  Benef Microbes     Publication Date:  2011 Mar 
Date Detail:
Created Date:  2011-08-11     Completed Date:  -     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  101507616     Medline TA:  Benef Microbes     Country:  Netherlands    
Other Details:
Languages:  eng     Pagination:  47-55     Citation Subset:  IM    
Affiliation:
Department of Arctic and Marine Biology, University of Tromsø, 9037 Tromsø, Norway.
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