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. 1988 Jun;170(6):2612–2617. doi: 10.1128/jb.170.6.2612-2617.1988

Cloning, nucleotide sequence, and taxonomic implications of the flagellin gene of Roseburia cecicola.

J H Martin 1, D C Savage 1
PMCID: PMC211178  PMID: 3372477

Abstract

The gene coding for the flagellin protein of Roseburia cecicola, an oxygen-intolerant, gram-negative, anaerobic bacterium indigenous to the murine cecum, has been cloned and sequenced. NH2-terminal amino acid sequence data from the flagellin protein were used as a basis for the synthesis of two mixed-sequence deoxyoligonucleotides. The oligonucleotides were used to identify and clone the flagellin structural gene. DNA sequence analysis of M13mp8 and mp9 subclones revealed a protein with a length of 293 amino acids and a molecular weight of 31,370. Comparisons with the sequences of flagellins of other species revealed conserved regions and suggested that although R. cecicola has structural characteristics of a gram-negative bacterium, it may be most closely related to the gram-positive bacteria.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Birnboim H. C., Doly J. A rapid alkaline extraction procedure for screening recombinant plasmid DNA. Nucleic Acids Res. 1979 Nov 24;7(6):1513–1523. doi: 10.1093/nar/7.6.1513. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Chet I., Mitchell R. Ecological aspects of microbial chemotactic behavior. Annu Rev Microbiol. 1976;30:221–239. doi: 10.1146/annurev.mi.30.100176.001253. [DOI] [PubMed] [Google Scholar]
  3. DeLange R. J., Chang J. Y., Shaper J. H., Glazer A. N. Amino acid sequence of flagellin of Bacillus subtilis 168. III. Tryptic peptides, N-bromosuccinimide peptides, and the complete amino acid sequence. J Biol Chem. 1976 Feb 10;251(3):705–711. [PubMed] [Google Scholar]
  4. Dretzen G., Bellard M., Sassone-Corsi P., Chambon P. A reliable method for the recovery of DNA fragments from agarose and acrylamide gels. Anal Biochem. 1981 Apr;112(2):295–298. doi: 10.1016/0003-2697(81)90296-7. [DOI] [PubMed] [Google Scholar]
  5. Gill P. R., Agabian N. A comparative structural analysis of the flagellin monomers of Caulobacter crescentus indicates that these proteins are encoded by two genes. J Bacteriol. 1982 May;150(2):925–933. doi: 10.1128/jb.150.2.925-933.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Gordon J. H., Dubos R. The anaerobic bacterial flora of the mouse cecum. J Exp Med. 1970 Aug 1;132(2):251–260. doi: 10.1084/jem.132.2.251. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Homma M., Fujita H., Yamaguchi S., Iino T. Regions of Salmonella typhimurium flagellin essential for its polymerization and excretion. J Bacteriol. 1987 Jan;169(1):291–296. doi: 10.1128/jb.169.1.291-296.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Joys T. M., Rankis V. The primary structure of the phase-1 flagellar protein of Salmonella typhimurium. I. The tryptic peptides. J Biol Chem. 1972 Aug 25;247(16):5180–5193. [PubMed] [Google Scholar]
  9. Kuwajima G., Asaka J., Fujiwara T., Fujiwara T., Node K., Kondo E. Nucleotide sequence of the hag gene encoding flagellin of Escherichia coli. J Bacteriol. 1986 Dec;168(3):1479–1483. doi: 10.1128/jb.168.3.1479-1483.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Logan S. M., Harris L. A., Trust T. J. Isolation and characterization of Campylobacter flagellins. J Bacteriol. 1987 Nov;169(11):5072–5077. doi: 10.1128/jb.169.11.5072-5077.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Logan S. M., Trust T. J. Location of epitopes on Campylobacter jejuni flagella. J Bacteriol. 1986 Nov;168(2):739–745. doi: 10.1128/jb.168.2.739-745.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Martin J. H., Savage D. C. Purification and characterization of flagella from Roseburia cecicola, an obligately anaerobic bacterium. J Gen Microbiol. 1985 Aug;131(8):2075–2078. doi: 10.1099/00221287-131-8-2075. [DOI] [PubMed] [Google Scholar]
  13. Messing J. New M13 vectors for cloning. Methods Enzymol. 1983;101:20–78. doi: 10.1016/0076-6879(83)01005-8. [DOI] [PubMed] [Google Scholar]
  14. Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Savage D. C. Microbial ecology of the gastrointestinal tract. Annu Rev Microbiol. 1977;31:107–133. doi: 10.1146/annurev.mi.31.100177.000543. [DOI] [PubMed] [Google Scholar]
  16. Simon M., Zieg J., Silverman M., Mandel G., Doolittle R. Phase variation: evolution of a controlling element. Science. 1980 Sep 19;209(4463):1370–1374. doi: 10.1126/science.6251543. [DOI] [PubMed] [Google Scholar]
  17. Southern E. M. Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol. 1975 Nov 5;98(3):503–517. doi: 10.1016/s0022-2836(75)80083-0. [DOI] [PubMed] [Google Scholar]
  18. Stanton T. B., Savage D. C. Colonization of gnotobiotic mice by Roseburia cecicola, a motile, obligately anaerobic bacterium from murine ceca. Appl Environ Microbiol. 1983 May;45(5):1677–1684. doi: 10.1128/aem.45.5.1677-1684.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Stanton T. B., Savage D. C. Motility as a factor in bowel colonization by Roseburia cecicola, an obligately anaerobic bacterium from the mouse caecum. J Gen Microbiol. 1984 Jan;130(1):173–183. doi: 10.1099/00221287-130-1-173. [DOI] [PubMed] [Google Scholar]
  20. Wei L. N., Joys T. M. Covalent structure of three phase-1 flagellar filament proteins of Salmonella. J Mol Biol. 1985 Dec 20;186(4):791–803. doi: 10.1016/0022-2836(85)90397-3. [DOI] [PubMed] [Google Scholar]

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