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Brevundimonas mediterranea sp. nov., a non-stalked species from the Mediterranean Sea, by I. Fritz, C. Strömpl, D. I. Nikitin, A. M. Lysenko and W.-R. Abraham

International Journal of Systematic and Evolutionary Microbiology vol. 55, part 1, pp. 479 - 486

Supplementary Table A. Properties of strains investigated in this study and sources of isolation. [PDF] (20 KB)

Supplementary Fig. A. Cells of Brevundimonas sp. V4.BO.10^T (transmission electron microscopy, negative stain).

Supplementary Fig. B. SDS-PAGE analyses of cell lysates from selected Brevundimonas strains. Cells were grown in FWC + 1 % NaCl to mid-exponential phase, i.e. to an OD₅₉₀ of 0.7-0.9. A volume of 2 ml bacterial suspension of each culture was harvested by centrifugation and stored at -20 °C until further processing. Prior to analysis, the cell pellet was resuspended in 20 µl CCE buffer [50 mM Tris/HCl, pH 6.8, 100 mM DTT, 2 % (w/v) SDS, 10 % (v/v) glycerol and 0.1 % (w/v) bromophenol blue]. Lysis was achieved by boiling the suspension for 2 min. The volume of the lysate was adjusted to 200 µl with sterile bidistilled water. Aliquots of 10 µl of the diluted lysate was applied per lane on a 10 % (w/v) SDS-PAGE gel, which was prepared and run as described elsewhere (Sambrook & Russell, 2001). Abbreviations: B. aur., Brevundimonas aurantiaca; B. ves., Brevundimonas vesicularis, B. int., Brevundimonas intermedia, B. sp., Brevundimonas sp. OTU H.

Supplementary Fig. C. RAPD PCR analyses of genomic DNA from selected Brevundimonas strains. RAPD PCR [primer UBC155 (5'-CTGGCGGCTG-3'); PCR conditions, 45 cycles of 15 s at 95 °C, 60 s at 36 °C, 120 s at 72 °C] was conducted as described elsewhere (Welsh & McClelland, 1990; Martin-Kearly et al., 1994; Ziemke et al., 1997). Aliquots of 8.5 µl of each PCR product were applied to a 1.5 % (w/v) agarose gel. Following electrophoresis, the gel was stained with ethidium bromide. Species abbreviations are outlined in the legend to Supplementary Fig. B.

[PDF file of Figs A-C] (597 KB)

References

Martin-Kearly, J., Gow, J. A., Péloquin, M. & Greer, C. W. (1994). Numerical analysis and the application of random amplified polymorphic DNA polymerase chain reaction to the differentiation of Vibrio strains from a seasonally cold ocean. Can J Microbiol 40, 446-455.
Sambrook, J. & Russell, D. W. (2001). Molecular Cloning: a Laboratory Manual. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory Press.
Welsh, J. J. & McClelland, M. (1990). Fingerprinting genomes using PCR with arbitrary primers. Nucleic Acids Res 18, 7213-7218.
Ziemke, F., Brettar, I. & Höfle, M. G. (1997). Stability and diversity of the genetic structure of a Shewanella putrefaciens population in the water column of the central Baltic. Aquat Microb Ecol 13, 63-74.

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