1887

Abstract

Summary

The lipopolysaccharides (LPS) of the 10 species of the genus ) were extracted by the proteinase K method and their resolution compared by several methods of polyacrylamide gel electrophoresis (PAGE). These included sodium dodecyl sulphate (SDS)-PAGE with and without urea, polyacrylamide gradient gels and Tricine [N-Tris (hydroxymethyl) methyl glycine]-SDS-PAGE. The original discontinuous system showed good resolution of LPS from and and this was enhanced by urea; showed a typical ladder pattern associated with repeating polysaccharide units of the O side chains. The LPS profiles of the other species, including , were poorly resolved; the majority of components migrated with the leading edge of the wave front. The resolution of the LPS of these species was marginally improved with gradient gels but the majority of components were separated only within the regions of high polyacrylamide concentration. The Tricine-SDS system was consistently superior to the other methods, with excellent resolution of the LPS profiles of all species.

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1991-05-01
2022-01-19
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References

  1. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 1970; 227:680–685
    [Google Scholar]
  2. Hitchcock P. J., Brown. T. M.. Morphological heterogenicityamong Salmonella lipopolysaccharide chemotypes in silver-stained polyacrylamide gels. J Bacterial 1983; 154:269–277
    [Google Scholar]
  3. Livermore D. M., Pitt T. L. Dissociation of surface properties and “intrinsic” resistance to /5-lactams in Pseudomonas aerogi-nosa. J Med Microbiol 1986; 22:217–224
    [Google Scholar]
  4. Apicella M. A., Dudas K. C., Campagnari A., Rice P., Mylotte J. M., Murphy T. F. Antigenic heterogeneity of lipid A of Haemophilus influenzae. Infect Immun 1985; 50:9–14
    [Google Scholar]
  5. Weintraub A., Zahringer U., Lindberg A. A. Structural studies of the polysaccharide part of the cell wall lipopolysaccharide from Bacteroides fragilis NCTC 9343. Eur J Biochem 1985; 151:657–661
    [Google Scholar]
  6. Linko-Kettunen L, Arstila P., Jalkanen M. Monoclonal antibodies to Bacteroides fragilis lipopolysaccharide. J Clin Microbiol 1984; 20:519–524
    [Google Scholar]
  7. Maskell J. P., Shah H. N. The distribution of lipopolysaccharide chemotypes within the redefined genus Bacteroides. In Borriello S. P. (ed) Clinical and molecular aspect of anaerobes Petersfield: Wrightson Biomedical Publishing; 1990309–310
    [Google Scholar]
  8. Poxton I. R., Brown R. Immunochemistry of the surface carbohydrate antigens of Bacteroides fragilis and definition. of a common antigen. J Gen Microbiol 1986; 132:2475–2481
    [Google Scholar]
  9. Breeling J. L., Onderdonk A. B., Cisneros R. L., Kasper D. L. Bacteroides vulgatus outer membrane antigens associated with carrageenan-induced colitis in guinea pigs. Infect Immun 1988; 56:1754–1759
    [Google Scholar]
  10. Schägger H, von Jagow G. Tricine-sodium dodecyl-sulfate-polyacrylamide gel electrophoresis for the separation of proteins in the range from 1 to 100 Kda. Anal Biochem 1987; 166:368–379
    [Google Scholar]
  11. Lesse A. J., Campagnari A. A., Bittner W. E., Apicella M. A. Increased resolution of lipopolysaccharides and lipooligo-saccharides utilizing tricine-sodium dodecyl sulfate-polyacrylamide gel electrophoresis. J Immunol Methods 1990; 126:109–117
    [Google Scholar]
  12. Shah H. N., Collins M. D. Proposal to restrict the genus Bacteroides (Castellani and Chalmers) to Bacteriodes fragilis and closely related species. Int J Syst Bacteriol 1989; 39:85–87
    [Google Scholar]
  13. Tsai C.-M., Frasch C. E. A sensitive silver stain for detecting lipopolysaccharides in polyacrylamide gels. Anal Biochem 1982; 119:115–119
    [Google Scholar]
  14. Kropinski A. M., Berry D., Greenberg E. P. The basis of silver staining of bacterial lipopolysaccharides in polyacrylamide gels. Curr Microbiol 1986; 13:29–31
    [Google Scholar]
  15. Lindberg A. A., Weintraub A., Zahringer U., Rietschel E. T. Structure-activity relationships in lipopolysaccharides of Bacteroides fragilis. Rev Infect Dis 1990; 12: Suppl 2S133–S141
    [Google Scholar]
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