Group-specific 16S rRNA hybridization probes for determinative and community structure studies of Butyrivibrio fibrisolvens in the rumen

被引:44
作者
Forster, RJ [1 ]
Gong, JH [1 ]
Teather, RM [1 ]
机构
[1] AGR CANADA,CTR FOOD & ANIM RES,RES BRANCH,OTTAWA,ON K1A 0C6,CANADA
关键词
D O I
10.1128/AEM.63.4.1256-1260.1997
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
Oligonucleotide probes covering three phylogenetically defined groups of Butyrivibrio spp, were successfully designed and tested, The specificity of each probe was examined by hybridization to rRNAs from an assortment of B. fibrisolvens isolates as well as additional ruminal and nonruminal bacteria. The sensitivity of the hybridization method was determined by using one of the probes (probe 156), When RNA was extracted from a culture of OB156, the probe was able to detect target cells at densities as low as 10(4) cells/ml, When 10(4) or more target cells/ml were added to cattle rumen samples, detectable hybridization signals were obtained with 1,000 ng of total RNA loaded onto the nylon membrane, In contrast, the sensitivity was reduced to 10(6) target cells/ml at 100 ng of RNA per slot, The probes were used to type 19 novel Butyrivibrio isolates, The phylogenetic placement was confirmed by partial 16S rRNA gene sequencing, The use of the probes in community-based studies indicated that the Butyvrivibrio groups examined in this paper did not represent a significant portion of the bacterial 16S rRNA pool in the rumen of the cattle, sheep, and deer examined.
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收藏
页码:1256 / 1260
页数:5
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