Skip to main content
SpringerLink
Log in

Nucleotide sequence of the gene coding for non-proteolyticClostridium botulinum type B neurotoxin: Comparison with other clostridial neurotoxins

  • Published:
Current Microbiology Aims and scope Submit manuscript

Abstract

The neurotoxin gene of non-proteolyticClostridium botulinum type B (strain Eklund 17B) was cloned as a series of overlapping polymerase chain reaction (PCR) fragments generated with primers designed to conserved regions of published botulinal toxin (BoNT) sequences. The 3′ end of the gene was obtained by using primers designed to the determined sequence of non-proteolytic BoNT/B and a published downstream region of BoNT/B gene from a proteolytic strain. Translation of the nucleotide sequence derived from cloned PCR fragments demonstrated the toxin gene encodes a protein of 1291 amino acid residues. Comparative alignment of the derived BoNT/B sequence with those of other published botulinal neurotoxins revealed highest sequence relatedness with BoNT/B of proteolyticC. botulinum. The sequence identity between non-proteolytic and proteolytic BoNT/B was 97.7% for the light chain (corresponding to 10 amino acid changes) and 90.2% for the heavy chain (corresponding to 81 amino acid changes), with most differences occurring at the C-terminal end. A genealogical tree constructed from all known botulinal neurotoxin sequences revealed marked topological differences with a phylogenetic tree ofC. botulinum types based upon small-subunit (16S) ribosomal RNA sequences.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

Literature Cited

  1. Aureli P, Fenicia L, Pasolini B, Gianfranceschi M, McCroskey LM, Hatheway CL (1986) Two cases of type E infant botulism caused by neurotoxigenicClostridium butyricum in Italy. J Infect Dis 154:207–211

    PubMed  Google Scholar 

  2. Binz T, Kurazono H, Popoff MR, Eklund MW, Sakaguchi G, Kozaki S, Krieglstein K, Henschen A, Gill DM, Neimann H (1990) Nucleotide sequence of the gene encodingClostridium botulinum neurotoxin type D. Nucleic Acids Res 18:5556

    PubMed  Google Scholar 

  3. Cato EP, George WL, Finegold SM (1986) GenusClostridium. In: Sneath PHA, Mair NS, Sharpe ME, Holt JG (eds) Bergey's manual of systematic bacteriology, vol 2. Baltimore: Williams and Wilkins, pp 1141–1200

    Google Scholar 

  4. Devereux J, Haeberli P, Smithies D (1984) A comprehensive set of sequence programs for the VAX. Nucleic Acids Res 12:387–395

    PubMed  Google Scholar 

  5. East AK, Richardson PT, Allaway D, Collins MD, Roberts TA, Thompson DE (1992) Sequence of the gene encoding type F neurotoxin ofClostridium botulinum. FEMS Microbiol Lett 96:225–230

    Google Scholar 

  6. Fairweather NF, Lyness VA (1986) The complete nucleotide sequence of tetanus toxin. Nucleic Acids Res 14:7809–7812

    PubMed  Google Scholar 

  7. Farrow JAE, Jones D, Phillips BA, Collins MD (1983) Taxonomic studies on some group D streptococci. J Gen Microbiol 129:1423–1432

    PubMed  Google Scholar 

  8. Felsenstein J (1989) PHYLIP-phylogeny inference package (version 3.2). Cladistics 5:64–166

    Google Scholar 

  9. Hall JD, McCroskey LM, Pincomb BJ, Hatheway CL (1985) Isolation of an organism resemblingClostridium baratii which produces type F botulinum toxin from an infant with botulism. J Clin Microbiol 21:654–655

    PubMed  Google Scholar 

  10. Hatheway CL (1992)Clostridium botulinum and other clostridia that produce botulinum neurotoxin. In: Hauschild AHW, Dodds KL (eds)Clostridium botulinum-ecology and control in foods. New York: Marcel Dekker Inc., pp 3–20

    Google Scholar 

  11. Hauser D, Eklund MW, Kurazono H, Binz T, Niemann H, Gill DM, Boquet P, Popoff MR (1990) Nucleotide sequence ofClostridium botulinum C1 neurotoxin. Nucleic Acids Res 18:4924

    PubMed  Google Scholar 

  12. Hutson RA, Thompson DE, Collins MD (1993a) Genetic interrelationships of saccharolyticClostridium botulinum types B, E and F and related clostridia as revealed by small-subunit rRNA gene sequences. FEMS Microbiol Lett 108:103–110

    PubMed  Google Scholar 

  13. Hutson RA, Thompson DE, Lawson PA, Schocken-Itturino RP, Bottger EC, Collins MD (1993b) Genetic interrelationships of proteolyticClostridium botulinum types A, B, and F and other members of theClostridium botulinum complex as revealed by small-subunit rRNA gene sequences. Antonie van Leeuwenhoek, in press

  14. Johnson JL, Francis BS (1975) Taxonomy of the clostridia: ribosomal ribonucleic acid homologies among the species. J Gen Microbiol 88:229–244

    PubMed  Google Scholar 

  15. Kautter DA, Solomon HM (1977) Collaborative studies of a method for the detection ofClostridium botulinum and its toxins in food. J Assoc Off Anal Chem 60:541–545

    PubMed  Google Scholar 

  16. Kozaki S, Miyazaki S, Sakaguchi G (1977) Development of antitoxin with each of two complementary fragments ofClostridium botulinum type B derivative toxin. Infect Immun 18:761–766

    PubMed  Google Scholar 

  17. McCroskey LM, Hatheway CL, Fenicia L, Pasolini B, Aureli P (1986) Characterization of an organism that produces type E botulinal toxin but which resemblesClostridium butyricum from the feces of an infant with type E botulism. J Clin Microbiol 23:201–202

    PubMed  Google Scholar 

  18. McCroskey LM, Hatheway CL, Woodruff BA, Greenberg JA, Jurgenson PA (1991) Type F botulism due to neurotoxigenicClostridium baratii from an unknown source in an adult. J Clin Microbiol 29:2618–2620

    PubMed  Google Scholar 

  19. Poulet S, Hauser D, Quanz M, Niemann H, Popoff MR (1992) Sequences of the botulinal neurotoxin derived fromClostridium botulinum type E (strain Beluga) andClostridium butyricum (strains ATCC 43181 and ATCC 43755). Biochem Biophys Res Commun 183:107–113

    PubMed  Google Scholar 

  20. Sambrook J, Fritsch EF, Maniatis T (1989) Molecular cloning, a laboratory manual, 2nd ed. Cold Spring Harbor, N.Y.: Cold Spring Harbor Laboratory Press

    Google Scholar 

  21. Thompson DE, Brehm JK, Oultram JD, Swinfield TJ, Shone CC, Atkinson T, Melling J, Minton NP (1990) The complete amino acid sequence ofClostridium botulinum type A neurotoxin, deduced by nucleotide sequence analysis of the encoding gene. Eur J Biochem 189:73–81

    PubMed  Google Scholar 

  22. Thompson DE, Hutson RA, East AK, Allaway D, Collins MD, Richardson PT (1993) Nucleotide sequence of the gene coding forClostridium baratii type F neurotoxin: comparison with other clostridial neurotoxins. FEMS Microbiol Lett 108:175–182

    PubMed  Google Scholar 

  23. Whelan SM, Elmore MJ, Bodsworth NJ, Brehm JK, Atkinson T, Minton NP (1992a) Molecular cloning of theClostridium botulinum structural gene encoding the type B neurotoxin and determination of its entire nucleotide sequence. Appl Environ Microbiol 58:2345–2354

    PubMed  Google Scholar 

  24. Whelan SM, Elmore MJ, Bodsworth NJ, Atkinson T, Minton NP (1992b) The complete amino acid sequence of theClostridium botulinum type E neurotoxin, derived by nucleotidesequence analysis of the encoding gene. Eur J Biochem 204:657–667

    PubMed  Google Scholar 

Download references

Author information

Author notes

  1. Daphne E. Thompson

    Present address: Department of Health, Skipton House, 80 London Road, SE1 6LW, London, UK

Authors and Affiliations

  1. Department of Microbiology, AFRC Institute of Food Research, Reading Laboratory, Earley Gate, Whiteknights Road, RG6 2EF, Reading, UK

    Roger A. Hutson, Matthew D. Collins, Alison K. East & Daphne E. Thompson

Authors
  1. Roger A. Hutson
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Matthew D. Collins
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Alison K. East
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Daphne E. Thompson
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Hutson, R.A., Collins, M.D., East, A.K. et al. Nucleotide sequence of the gene coding for non-proteolyticClostridium botulinum type B neurotoxin: Comparison with other clostridial neurotoxins. Current Microbiology 28, 101–110 (1994). https://doi.org/10.1007/BF01569055

Download citation

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF01569055

Keywords

Search

Navigation