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Applied and Environmental Microbiology, July 1999, p. 3129-3133, Vol. 65, No. 7
0099-2240/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

Identification and Characterization of a Flagellin Gene from the Endosymbiont of the Hydrothermal Vent Tubeworm Riftia pachyptila

Deborah S. Millikan,dagger Horst Felbeck, and Jeffrey L. Stein*

Marine Biology Research Division, Scripps Institution of Oceanography, University of California, San Diego, La Jolla, California 92093-0202

Received 30 December 1998/Accepted 16 April 1999

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 sigma 28 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.

* Corresponding author. Present address: Quorum Pharmaceuticals, 13525 Samantha Ave., San Diego, CA 92129. Phone: (619) 538-5780. Fax: (619) 538-5009. E-mail: jstein{at}qpharm.com.

dagger Formerly published as D. S. Hughes.

Applied and Environmental Microbiology, July 1999, p. 3129-3133, Vol. 65, No. 7
0099-2240/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


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