Abstract
The intron-encoded endonuclease I-CeuI fromChlamydomonas eugametos was shown to cleave the circular chromosomes of allClostridium perfringens strains examined at single sites in the rRNA operons, thereby generating ten fragments suitable for the rapid mapping of virulence genes by pulsed-field gel electrophoresis (PFGE). This method easily distinguishes between plasmid and chromosomal localisations, as I-CeuI only cuts chromosomal DNA. Using this approach, the genes for three of the four typing toxins,β, ε, andı, in addition to the enterotoxin andλ-toxin genes, were shown to be plasmid-borne. In a minority of strains, associated with food poisoning, where the enterotoxin toxin gene was located on the chromosome, genes for two of the minor toxins,ϑ andμ, were missing.
This is a preview of subscription content, log in via an institution to check access.
References
Canard B, Cole ST (1989) Genome organization of the anaerobic pathogenClostridium perfringens. Proc Natl Acad Sci USA 86:6676–6680
Canard B, Saint-Joanis B, Cole ST (1992) Genomic diversity and organisation of virulence genes in the pathogenic anaerobeClostridium perfringens. Mol Microbiol 6:1421–1429
Cornillot E, Saint-Joanis B, Daube G, Katayama S, Granum PE, Canard B, Cole ST (1995) The enterotoxin gene (cpe) ofClostridium perfringens can be chromosomal or plasmid-borne. Mol Microbiol 15:639–647
Daube G, Simon P, Kaeckenbeeck A (1993) IS1151, an IS-like element ofClostridium perfringens. Nucleic Acids Res 21:352
Daube G, China B, Simon P, Hvala K, Mainil J (1994) Typing ofClostridium perfringens by in vitro amplification of toxin genes. J Appl Bacteriol 77:650–655
Duncan CL, Rokos EA, Christenson CM, Rood JI (1978) Multiple plasmids in different toxigenic strains ofClostridium perfringens: possible control of beta toxin production. In: Schlessinger D (ed) Microbiology 1978. American Society for Mirobiology, Washington, DC, pp 246–248
Eklund MW (1992) The role of bacteriophages and plasmids in the production of toxins and other biologically active substances byClostridium botulinum andClostridium novyi. In: Sebald M (ed) Genetics and molecular biology of anaerobic bacteria. Springer-Verlag, New York, pp 179–194
Finn CW, Silver RP, Habig WH, Hardegree MC (1984) The structural gene for tetanus toxin is on a plasmid. Science 224:881–884
Granum P (1990)Clostridium perfringens toxins involved in food poisoning. Int J Food Microbiol 10:101–112
Hatheway CL (1990) Toxigenic Clostridia. Clin Microbiol Rev 3:66–98
Hunter SEC, Brown JE, Oyston PCF, Sakurai J, Titball RW (1993) Molecular genetic analysis of beta-toxin ofClostridium perfringens reveals sequence homology with alpha-toxin, gamma-toxin, and leukocidin ofStaphylococcus aureus. Infect Immun 61:3958–3965
Jin F, Matsushita O, Katayama S-I, Jin S, Maatsushita C, Minami J, Okabe A (1996) Purification, characterization, and primary structure ofClostridium perfringens lambda-toxin, a thermolysin-like metalloprotease. Infect Immun 64:230–237
Katayama S, Dupuy B, Cole ST (1995) Rapid expansion of the physical and genetic map of the chromosome ofClostridium perfringens CPN50. J Bacteriol 177:5680–5685
Marshall P, Lemieux C (1991) Cleavage pattern of the homing endonuclease encoded by the fifth intron in the chloroplast large subunit rRNA-encoding gene ofChlamydomonas eugametos. Gene 104:241–245
McDonel JL (1986) Toxins ofClostridium perfringens types A, B, C, D and E. In: Dorner F, Drews J (eds) Pharmacology of bacterial toxins. Pergamon Press, Oxford, pp 477–517
Perelle S, Gibert M, Boquet P, Popoff M (1993) Characterization ofClostridium perfringens iota-toxin genes and expression inEscherichia coli. Infect Immun 61:5147–5156
Rood JI, Cole ST (1991) Molecular genetics and pathogenesis ofClostridium perfringens. Microbiol Rev 55:621–648
Smith CL, Warburton PE, Gaal A, Cantor CR (1986) Analysis of genome organization and rearrangements by pulsed field gradient gel electrophoresis. In: Setlow JK, Hollaender A (eds) Genetic engineering. Plenum, New York, pp 45–70
Author information
Authors and Affiliations
Additional information
Communicated by A. Kondorosi
Rights and permissions
About this article
Cite this article
Katayama, S., Dupuy, B., Cole, S.T. et al. Genome mapping ofClostridium perfringens strains with I-CeuI shows many virulence genes to be plasmid-borne. Molec. Gen. Genet. 251, 720–726 (1996). https://doi.org/10.1007/BF02174122
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF02174122