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. 1971 Jan;105(1):376–383. doi: 10.1128/jb.105.1.376-383.1971

Purification of Intact Flagella from Escherichia coli and Bacillus subtilis

M L DePamphilis 1, Julius Adler 2
PMCID: PMC248365  PMID: 4993324

Abstract

A procedure is described for the purification of bacterial flagella in the form of a filament-hook-basal body complex (intact flagella) free from detectable cell wall, membrane, or cytoplasmic material. Spheroplasts produced with lysozyme and ethylenediaminetetraacetic acid were lysed with Triton X-100, and the flagella were purified by (NH4)2SO4 precipitation, differential centrifugation, and CsCl gradient centrifugation. As much as 40% of the flagella were recovered, and they contained about one basal body per 4 to 6 μm of flagella. The same procedure developed for Escherichia coli was also successful for purifying intact flagella from Bacillus subtilis.

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Selected References

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

  1. ABRAM D., KOFFLER H. IN VITRO FORMATION OF FLAGELLA-LIKE FILAMENTS AND OTHER STRUCTURES FROM FLAGELLIN. J Mol Biol. 1964 Jul;9:168–185. doi: 10.1016/s0022-2836(64)80098-x. [DOI] [PubMed] [Google Scholar]
  2. Abram D., Koffler H., Vatter A. E. Basal structure and attachment of flagella in cells of Proteus vulgaris. J Bacteriol. 1965 Nov;90(5):1337–1354. doi: 10.1128/jb.90.5.1337-1354.1965. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Abram D., Mitchen J. R., Koffler H., Vatter A. E. Differentiation within the bacterial flagellum and isolation of the proximal hook. J Bacteriol. 1970 Jan;101(1):250–261. doi: 10.1128/jb.101.1.250-261.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Armstrong J. B., Adler J. Genetics of motility in Escherichia coli: complementation of paralysed mutants. Genetics. 1967 Jul;56(3):363–373. doi: 10.1093/genetics/56.3.363. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Cohen-Bazire G., London J. Basal organelles of bacterial flagella. J Bacteriol. 1967 Aug;94(2):458–465. doi: 10.1128/jb.94.2.458-465.1967. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. DAVIS B. J. DISC ELECTROPHORESIS. II. METHOD AND APPLICATION TO HUMAN SERUM PROTEINS. Ann N Y Acad Sci. 1964 Dec 28;121:404–427. doi: 10.1111/j.1749-6632.1964.tb14213.x. [DOI] [PubMed] [Google Scholar]
  7. DePamphilis M. L., Adler J. Attachment of flagellar basal bodies to the cell envelope: specific attachment to the outer, lipopolysaccharide membrane and the cyoplasmic membrane. J Bacteriol. 1971 Jan;105(1):396–407. doi: 10.1128/jb.105.1.396-407.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. DePamphilis M. L., Adler J. Fine structure and isolation of the hook-basal body complex of flagella from Escherichia coli and Bacillus subtilis. J Bacteriol. 1971 Jan;105(1):384–395. doi: 10.1128/jb.105.1.384-395.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Dimmitt K., Simon M. Antigenic nature of bacterial flagellar hook structures. Infect Immun. 1970 Feb;1(2):212–213. doi: 10.1128/iai.1.2.212-213.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Dimmitt K., Simon M. Purification and thermal stability of intact Bacillus subtilis flagella. J Bacteriol. 1971 Jan;105(1):369–375. doi: 10.1128/jb.105.1.369-375.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Doetsch R. N., Hageage G. J. Motility in procaryotic organisms: problems, points of view, and perspectives. Biol Rev Camb Philos Soc. 1968 Aug;43(3):317–362. doi: 10.1111/j.1469-185x.1968.tb00963.x. [DOI] [PubMed] [Google Scholar]
  12. ENGLANDER S. W., EPSTEIN H. T. Optical methods for measuring nucleoprotein and nucleic acid concentrations. Arch Biochem Biophys. 1957 May;68(1):144–149. doi: 10.1016/0003-9861(57)90334-x. [DOI] [PubMed] [Google Scholar]
  13. Grant G. F., Simon M. Use of radioactive antibodies for characterizing antigens and application to the study of flagella synthesis. J Bacteriol. 1968 Jan;95(1):81–86. doi: 10.1128/jb.95.1.81-86.1968. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Joys T. M. The structure of flagella and the genetic control of flagellation in Eubacteriales. A review. Antonie Van Leeuwenhoek. 1968;34(2):205–225. doi: 10.1007/BF02046432. [DOI] [PubMed] [Google Scholar]
  15. Lawn A. M. Simple immunological labelling method for electron microscopy and its application to the study of filamentous appendages of bacteria. Nature. 1967 Jun 10;214(5093):1151–1152. doi: 10.1038/2141151a0. [DOI] [PubMed] [Google Scholar]
  16. McConahey P. J., Dixon F. J. A method of trace iodination of proteins for immunologic studies. Int Arch Allergy Appl Immunol. 1966;29(2):185–189. doi: 10.1159/000229699. [DOI] [PubMed] [Google Scholar]
  17. REISFELD R. A., LEWIS U. J., WILLIAMS D. E. Disk electrophoresis of basic proteins and peptides on polyacrylamide gels. Nature. 1962 Jul 21;195:281–283. doi: 10.1038/195281a0. [DOI] [PubMed] [Google Scholar]
  18. REPASKE R. Lysis of gram-negative organisms and the role of versene. Biochim Biophys Acta. 1958 Nov;30(2):225–232. doi: 10.1016/0006-3002(58)90044-1. [DOI] [PubMed] [Google Scholar]
  19. STOCKER B. A., CAMPBELL J. C. The effect of non-lethal deflagellation on bacterial motility and observations on flagellar regeneration. J Gen Microbiol. 1959 Jun;20(3):670–685. doi: 10.1099/00221287-20-3-670. [DOI] [PubMed] [Google Scholar]
  20. Vaituzis Z., Doetsch R. N. Relationship between cell wall, cytoplasmic membrane, and bacterial motility. J Bacteriol. 1969 Oct;100(1):512–521. doi: 10.1128/jb.100.1.512-521.1969. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Van Iterson W., Hoeniger J. F., Van Zanten E. N. Basal bodies of bacterial flagella in Proteus mirabilis. I. Electron microscopy of sectioned material. J Cell Biol. 1966 Dec;31(3):585–601. doi: 10.1083/jcb.31.3.585. [DOI] [PMC free article] [PubMed] [Google Scholar]

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