Sequence Analysis of 16S RRNA from Mycoplasmas by Direct Solid-phase DNA Sequencing

Overview

Journal Appl Environ Microbiol

Specialties Environmental Health
Microbiology

Date 1994 Jul 1

PMID 7521158

Citations 16

Authors

B Pettersson

K E Johansson

M Uhlen

Affiliations

Soon will be listed here.

Abstract

Automated solid-phase DNA sequencing was used for determination of partial 16S ribosomal DNA sequences of mycoplasmas. The sequence information was used to establish phylogenetic relationships of 11 different mycoplasmas whose 16S rRNA sequences had not been determined earlier. A biotinylated fragment corresponding to positions 344 to 939 in the Escherichia coli sequence was generated by PCR. The PCR product was immobilized onto streptavidin-coated paramagnetic beads, and direct sequencing was performed in both directions. One previously unclassified avian mycoplasma was found to belong to the Mycoplasma lipophilum cluster of the hominis group. Microheterogeneities were discovered in the rRNA operons of Mycoplasma mycoides subsp. mycoides (SC type), confirming the existence of two different rRNA operons. The 16S rRNA sequence of M. mycoides subsp. capri was identical to that of M. mycoides subsp. mycoides (type SC), except that no microheterogeneities were revealed. Furthermore, automated solid-phase DNA sequencing was used to identify a mycoplasmal contamination of a cell culture as Mycoplasma hyorhinis, which proved to be very difficult by conventional methods. The results suggest that the direct solid-phase DNA sequencing procedure is a powerful tool for identification of mycoplasmas and is also useful in taxonomic studies.

Citing Articles

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Mycoplasma hyopharyngis isolation from swine.

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Spatial and resource factors influencing high microbial diversity in soil.

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Impact of dilution on microbial community structure and functional potential: comparison of numerical simulations and batch culture experiments.

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Diagnosis of contagious bovine pleuropneumonia by PCR-laser- induced fluorescence and PCR-restriction endonuclease analysis based on the 16S rRNA genes of Mycoplasma mycoides subsp. mycoides SC.

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