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. 1983 Jul;41(1):54–60. doi: 10.1128/iai.41.1.54-60.1983

Cloning and analysis of the K1 capsule biosynthesis genes of Escherichia coli: lack of homology with Neisseria meningitidis group B DNA sequences.

C Echarti, B Hirschel, G J Boulnois, J M Varley, F Waldvogel, K N Timmis
PMCID: PMC264742  PMID: 6408005

Abstract

Genes coding for production of the K1 polysaccharide capsule of Escherichia coli have been cloned. Complementation, insertion, and deletion analyses were used to localize the K1 genes and demonstrated that a minimum of 9 kilobases of DNA split into at least two gene blocks is involved in synthesis and assembly of the capsule. One of the gene blocks is responsible for biosynthesis of the polysaccharide, and the other is responsible for extracellular appearance of capsular material. Using cloned K1 genes as probes in Southern blot experiments, we detected homology to DNA from strains of E. coli capsular types K92, K7, and K100. In contrast, no homology was apparent between K1 genes and DNA from meningococcus group B, although the K1 and group B capsules are chemically and immunologically identical.

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Selected References

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  1. Andrés I., Slocombe P. M., Cabello F., Timmis J. K., Lurz R., Burkardt H. J., Timmis K. N. Plasmid replication functions. II. Cloning analysis of the repA replication region of antibiotic resistance plasmid R6-5. Mol Gen Genet. 1979 Jan 5;168(1):1–25. doi: 10.1007/BF00267929. [DOI] [PubMed] [Google Scholar]
  2. Chang A. C., Cohen S. N. Construction and characterization of amplifiable multicopy DNA cloning vehicles derived from the P15A cryptic miniplasmid. J Bacteriol. 1978 Jun;134(3):1141–1156. doi: 10.1128/jb.134.3.1141-1156.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Collins J., Hohn B. Cosmids: a type of plasmid gene-cloning vector that is packageable in vitro in bacteriophage lambda heads. Proc Natl Acad Sci U S A. 1978 Sep;75(9):4242–4246. doi: 10.1073/pnas.75.9.4242. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Egan W., Liu T. Y., Dorow D., Cohen J. S., Robbins J. D., Gotschlich E. C., Robbins J. B. Structural studies on the sialic acid polysaccharide antigen of Escherichia coli strain Bos-12. Biochemistry. 1977 Aug 9;16(16):3687–3692. doi: 10.1021/bi00635a028. [DOI] [PubMed] [Google Scholar]
  5. Franklin F. C., Bagdasarian M., Bagdasarian M. M., Timmis K. N. Molecular and functional analysis of the TOL plasmid pWWO from Pseudomonas putida and cloning of genes for the entire regulated aromatic ring meta cleavage pathway. Proc Natl Acad Sci U S A. 1981 Dec;78(12):7458–7462. doi: 10.1073/pnas.78.12.7458. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Gross R. J., Cheasty T., Rowe B. Isolation of bacteriophages specific for the K1 polysaccharide antigen of Escherichia coli. J Clin Microbiol. 1977 Dec;6(6):548–550. doi: 10.1128/jcm.6.6.548-550.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Guyer M. S. The gamma delta sequence of F is an insertion sequence. J Mol Biol. 1978 Dec 15;126(3):347–365. doi: 10.1016/0022-2836(78)90045-1. [DOI] [PubMed] [Google Scholar]
  8. Hohn B. In vitro packaging of lambda and cosmid DNA. Methods Enzymol. 1979;68:299–309. doi: 10.1016/0076-6879(79)68021-7. [DOI] [PubMed] [Google Scholar]
  9. Holmes D. S., Quigley M. A rapid boiling method for the preparation of bacterial plasmids. Anal Biochem. 1981 Jun;114(1):193–197. doi: 10.1016/0003-2697(81)90473-5. [DOI] [PubMed] [Google Scholar]
  10. Horwitz M. A., Silverstein S. C. Influence of the Escherichia coli capsule on complement fixation and on phagocytosis and killing by human phagocytes. J Clin Invest. 1980 Jan;65(1):82–94. doi: 10.1172/JCI109663. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Jeffreys A. J., Flavell R. A. A physical map of the DNA regions flanking the rabbit beta-globin gene. Cell. 1977 Oct;12(2):429–439. doi: 10.1016/0092-8674(77)90119-2. [DOI] [PubMed] [Google Scholar]
  12. Jorgensen R. A., Rothstein S. J., Reznikoff W. S. A restriction enzyme cleavage map of Tn5 and location of a region encoding neomycin resistance. Mol Gen Genet. 1979;177(1):65–72. doi: 10.1007/BF00267254. [DOI] [PubMed] [Google Scholar]
  13. Legerski R. J., Hodnett J. L., Gray H. B., Jr Extracellular nucleases of pseudomonas BAL 31. III. Use of the double-strand deoxyriboexonuclease activity as the basis of a convenient method for the mapping of fragments of DNA produced by cleavage with restriction enzymes. Nucleic Acids Res. 1978 May;5(5):1445–1464. doi: 10.1093/nar/5.5.1445. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Lurz R., Danbara H., Rückert B., Timmis K. N. Plasmid replication functions. VII. Electron microscopic localization of RNA polymerase binding sites in the replication control region of plasmid R6-5. Mol Gen Genet. 1981;183(3):490–496. doi: 10.1007/BF00268770. [DOI] [PubMed] [Google Scholar]
  15. McCracken G. H., Jr, Sarff L. D., Glode M. P., Mize S. G., Schiffer M. S., Robbins J. B., Gotschlich E. C., Orskov I., Orskov F. Relation between Escherichia coli K1 capsular polysaccharide antigen and clinical outcome in neonatal meningitis. Lancet. 1974 Aug 3;2(7875):246–250. doi: 10.1016/s0140-6736(74)91413-5. [DOI] [PubMed] [Google Scholar]
  16. Orskov I., Orskov F., Jann B., Jann K. Serology, chemistry, and genetics of O and K antigens of Escherichia coli. Bacteriol Rev. 1977 Sep;41(3):667–710. doi: 10.1128/br.41.3.667-710.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Peterson P. K., Quie P. G. Bacterial surface components and the pathogenesis of infectious diseases. Annu Rev Med. 1981;32:29–43. doi: 10.1146/annurev.me.32.020181.000333. [DOI] [PubMed] [Google Scholar]
  18. Pitt J. K-1 antigen of Escherichia coli: epidemiology and serum sensitivity of pathogenic strains. Infect Immun. 1978 Oct;22(1):219–224. doi: 10.1128/iai.22.1.219-224.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. SAITO H., MIURA K. I. PREPARATION OF TRANSFORMING DEOXYRIBONUCLEIC ACID BY PHENOL TREATMENT. Biochim Biophys Acta. 1963 Aug 20;72:619–629. [PubMed] [Google Scholar]
  20. Silver R. P., Finn C. W., Vann W. F., Aaronson W., Schneerson R., Kretschmer P. J., Garon C. F. Molecular cloning of the K1 capsular polysaccharide genes of E. coli. Nature. 1981 Feb 19;289(5799):696–698. doi: 10.1038/289696b0. [DOI] [PubMed] [Google Scholar]
  21. Smith H. Microbial surfaces in relation to pathogenicity. Bacteriol Rev. 1977 Jun;41(2):475–500. doi: 10.1128/br.41.2.475-500.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. 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]
  23. Timmis K. N., Cabello F., Cohen S. N. Cloning and characterization of EcoRI and HindIII restriction endonuclease-generated fragments of antibiotic resistance plasmids R6-5 and R6. Mol Gen Genet. 1978 Jun 14;162(2):121–137. doi: 10.1007/BF00267869. [DOI] [PubMed] [Google Scholar]
  24. Wilfert C. M. E. coli meningitis: K1 antigen and virulence. Annu Rev Med. 1978;29:129–136. doi: 10.1146/annurev.me.29.020178.001021. [DOI] [PubMed] [Google Scholar]
  25. Zwahlen A., Nydegger U. E., Vaudaux P., Lambert P. H., Waldvogel F. A. Complement-mediated opsonic activity in normal and infected human cerebrospinal fluid: early response during bacterial meningitis. J Infect Dis. 1982 May;145(5):635–646. doi: 10.1093/infdis/145.2.635. [DOI] [PubMed] [Google Scholar]

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