Aquatic Sciences Meeting, Albuquerque 2001

SS14 Microbial Diversity (Disciplinary Connections)

Date: Monday, February 12, 2001, Time: 4:30:00 PM

Location: Brazos

MacGregor, B, J, Max Planck Institute for Marine Microbiology, Bremen, Germany, bmacgreg@mpi-bremen.de

Bruechert, V, , Max Planck Institute for Marine Microbiology, Bremen, Germany, vbrucher@mpi-bremen.de

Joergensen, B, B, Max Planck Institute for Marine Microbiology, Bremen, Germany, bjoergen@mpi-bremen.de

Amann, R, , Max Planck Institute for Marine Microbiology, Bremen, Germany, ramann@mpi-bremen.de

ISOLATION OF SPECIFIC 16S RIBOSOMAL RNAS FROM MIXED SAMPLES FOR STABLE CARBON ISOTOPE AND MOLECULAR BIOLOGICAL CHARACTERIZATION

One approach to investigating the physiology of microbes in natural assemblages relies on the isotopic fractionations associated with enzymatic processes, such as methanogenesis, which may result in production of biomarker compounds with distinct isotopic compositions. Small-subunit rRNA is one potential phylogenetically informative biomarker. We have developed a method for isolating specific rRNAs from mixed microbial populations, employing biotin-labeled oligonucleotide probes and streptavidin-coated paramagnetic beads. In addition to its potential usefulness for the characterization of microorganisms not yet in culture, this method offers a way to identify the microbes actually targeted by oligonucleotide probes. For example, we have shown that a widely-used probe (S-D-Arch-0915-a-A-20), highly specific for the archaea by sequence database comparison, also hybridizes efficiently to 16S rRNA from members of the Cytophaga/ Flavobacterium/ Bacteroides group. The magnetic bead method may also be useful for isolation of specific full-length 16S rRNA molecules from natural samples, without the length limitations imposed by the need for primer pairs in PCR.

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Aquatic Sciences Meeting, Albuquerque 2001

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