Identification and characterization of a flagellin gene from the endosymbiont of the hydrothermal vent tubeworm Riftia pachyptila

The bacterial endosymbionts of the hydrothermal vent tubeworm Riftia pachyptila play a key role in providing their host with fixed carbon. Results of prior research suggest that the symbionts are selected from an environmental bacterial population, although a free-living form has been neither cultur...

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Bibliographic Details
Published in:Applied and environmental microbiology 1999-07, Vol.65 (7), p.3129-3133
Main Authors: MILLIKAN, D. S, FELBECK, H, STEIN, J. L
Format: Article
Language:English
Subjects:
Animals
Bacteria
Bacteria - genetics
Bacteria - metabolism
Bacterial Physiological Phenomena
Bacteriology
Biological and medical sciences
Blotting, Western
Carbon
Escherichia coli - genetics
Escherichia coli - metabolism
Flagellin - genetics
Flagellin - isolation & purification
Flagellin - metabolism
Fundamental and applied biological sciences. Psychology
Genes, Bacterial
Genetics
Invertebrate Microbiology
Microbiology
Microscopy, Electron
Molecular Sequence Data
Movement
Polychaeta - microbiology
Riftia pachyptila
Symbiosis
Worms
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Summary:The bacterial endosymbionts of the hydrothermal vent tubeworm Riftia pachyptila play a key role in providing their host with fixed carbon. Results of prior research suggest that the symbionts are selected from an environmental bacterial population, although a free-living form has been neither cultured from nor identified in the hydrothermal vent environment. To begin to assess the free-living potential of the symbiont, we cloned and characterized a flagellin gene from a symbiont fosmid library. The symbiont fliC gene has a high degree of homology with other bacterial flagellin genes in the amino- and carboxy-terminal regions, while the central region was found to be nonconserved. A sequence that was homologous to that of a consensus sigma28 RNA polymerase recognition site lay upstream of the proposed translational start site. The symbiont protein was expressed in Escherichia coli, and flagella were observed by electron microscopy. A 30,000-Mr protein subunit was identified in whole-cell extracts by Western blot analysis. These results provide the first direct evidence of a motile free-living stage of a chemoautotrophic symbiont and support the hypothesis that the symbiont of R. pachyptila is acquired with each new host generation.
ISSN:0099-2240
1098-5336
DOI:10.1128/aem.65.7.3129-3133.1999