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Phylogenetic group-specific oligodeoxynucleotide probes for identification of single microbial cells.

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  • 1. Department of Biology, Indiana University, Bloomington 47405.
      Authors
    • Giovannoni SJ 1
    (1 author)

ORCIDs linked to this article

Journal of Bacteriology, 01 Feb 1988, 170(2):720-726
https://doi.org/10.1128/jb.170.2.720-726.1988 PMID: 2448289 PMCID: PMC210714

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This article has been corrected. See J Bacteriol 1988 May;170(5):2418.

Abstract 

Examination of collections of 16S rRNA sequences revealed sequence domains that were unique to (and invariant within) the three primary lines of cellular descent: the archaebacteria, the eubacteria, and the eucaryotes. Oligodeoxynucleotides complementary to these conserved sequence domains were synthesized and used as hybridization probes. Each of the radiolabeled probes specifically hybridized to nylon membrane-bound 16S rRNA from the targeted kingdom. A probe complementary to a universally conserved sequence in 16S rRNAs was used as a positive control, while its complement provided a negative control for nonspecific binding. The abilities of the probes to bind specifically to whole, fixed cells representing a broad array of phylogenetic diversity were tested in whole-cell dot blot assays. Again, all of the probes specifically bound the targeted groups. By microautoradiography, the method was extended to permit phylogenetic identification of single cells microscopically.

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Logo of jbacterLink to Publisher's site
J Bacteriol. 1988 Feb; 170(2): 720–726.
PMCID: PMC210714
PMID: 2448289

Phylogenetic group-specific oligodeoxynucleotide probes for identification of single microbial cells.

S J Giovannoni, E F DeLong, G J Olsen, and N R Pace
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Abstract

Examination of collections of 16S rRNA sequences revealed sequence domains that were unique to (and invariant within) the three primary lines of cellular descent: the archaebacteria, the eubacteria, and the eucaryotes. Oligodeoxynucleotides complementary to these conserved sequence domains were synthesized and used as hybridization probes. Each of the radiolabeled probes specifically hybridized to nylon membrane-bound 16S rRNA from the targeted kingdom. A probe complementary to a universally conserved sequence in 16S rRNAs was used as a positive control, while its complement provided a negative control for nonspecific binding. The abilities of the probes to bind specifically to whole, fixed cells representing a broad array of phylogenetic diversity were tested in whole-cell dot blot assays. Again, all of the probes specifically bound the targeted groups. By microautoradiography, the method was extended to permit phylogenetic identification of single cells microscopically.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.
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Funding 

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NIGMS NIH HHS (1)