Genetic analysis of type E botulinum toxin-producing Clostridium butyricum strains

X. Wang, T. Maegawa, T. Karasawa, S. Kozaki, Kentaro Tsukamoto, Y. Gyobu, K. Yamakawa, K. Oguma, Y. Sakaguchi, S. Nakamura

Research output: Contribution to journalArticle

50 Citations (Scopus)

Abstract

Type E botulinum toxin (BoNT/E)-producing Clostridium butyricum strains isolated from botulism cases or soil specimens in Italy and China were analyzed by using nucleotide sequencing of the bont/E gene, random amplified polymorphic DNA (RAPD) assay, pulsed-field gel electrophoresis (PFGE), and Southern blot hybridization for the bont/E gene. Nucleotide sequences of the bont/E genes of 11 Chinese isolates and of the Italian strain BL 6340 were determined. The nucleotide sequences of the bont/E genes of 11 C. butyricum isolates from China were identical. The deduced amino acid sequence of BoNT/E from the Chinese isolates showed 95.0 and 96.9% identity with those of BoNT/E from C. butyricum BL 6340 and Clostridium botulinum type E, respectively. The BoNT/E-producing C. butyricum strains were divided into the following three clusters based on the results of RAPD assay, PFGE profiles of genomic DNA digested with SmaI or XhoI, and Southern blot hybridization: strains associated with infant botulism in Italy, strains associated with food-borne botulism in China, and isolates from soil specimens of the Weishan lake area in China. A DNA probe for the bont/E gene hybridized with the nondigested chromosomal DNA of all toxigenic strains tested, indicating chromosomal localization of the bont/E gene in C. butyricum. The present results suggest that BoNT/E-producing C. butyricum is clonally distributed over a vast area.

Original languageEnglish
Pages (from-to)4992-4997
Number of pages6
JournalApplied and Environmental Microbiology
Volume66
Issue number11
DOIs
Publication statusPublished - 18-11-2000

Fingerprint

Clostridium butyricum
botulinum toxin
genetic analysis
toxin
genetic techniques and protocols
Botulism
botulism
gene
China
DNA
Genes
random amplified polymorphic DNA technique
genes
Pulsed Field Gel Electrophoresis
pulsed-field gel electrophoresis
Southern Blotting
Southern blotting
Italy
electrokinesis
Clostridium botulinum type E

All Science Journal Classification (ASJC) codes

  • Biotechnology
  • Food Science
  • Applied Microbiology and Biotechnology
  • Ecology

Cite this

Wang, X. ; Maegawa, T. ; Karasawa, T. ; Kozaki, S. ; Tsukamoto, Kentaro ; Gyobu, Y. ; Yamakawa, K. ; Oguma, K. ; Sakaguchi, Y. ; Nakamura, S. / Genetic analysis of type E botulinum toxin-producing Clostridium butyricum strains. In: Applied and Environmental Microbiology. 2000 ; Vol. 66, No. 11. pp. 4992-4997.
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Wang, X, Maegawa, T, Karasawa, T, Kozaki, S, Tsukamoto, K, Gyobu, Y, Yamakawa, K, Oguma, K, Sakaguchi, Y & Nakamura, S 2000, 'Genetic analysis of type E botulinum toxin-producing Clostridium butyricum strains', Applied and Environmental Microbiology, vol. 66, no. 11, pp. 4992-4997. https://doi.org/10.1128/AEM.66.11.4992-4997.2000

Genetic analysis of type E botulinum toxin-producing Clostridium butyricum strains. / Wang, X.; Maegawa, T.; Karasawa, T.; Kozaki, S.; Tsukamoto, Kentaro; Gyobu, Y.; Yamakawa, K.; Oguma, K.; Sakaguchi, Y.; Nakamura, S.

In: Applied and Environmental Microbiology, Vol. 66, No. 11, 18.11.2000, p. 4992-4997.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Genetic analysis of type E botulinum toxin-producing Clostridium butyricum strains

AU - Wang, X.

AU - Maegawa, T.

AU - Karasawa, T.

AU - Kozaki, S.

AU - Tsukamoto, Kentaro

AU - Gyobu, Y.

AU - Yamakawa, K.

AU - Oguma, K.

AU - Sakaguchi, Y.

AU - Nakamura, S.

PY - 2000/11/18

Y1 - 2000/11/18

N2 - Type E botulinum toxin (BoNT/E)-producing Clostridium butyricum strains isolated from botulism cases or soil specimens in Italy and China were analyzed by using nucleotide sequencing of the bont/E gene, random amplified polymorphic DNA (RAPD) assay, pulsed-field gel electrophoresis (PFGE), and Southern blot hybridization for the bont/E gene. Nucleotide sequences of the bont/E genes of 11 Chinese isolates and of the Italian strain BL 6340 were determined. The nucleotide sequences of the bont/E genes of 11 C. butyricum isolates from China were identical. The deduced amino acid sequence of BoNT/E from the Chinese isolates showed 95.0 and 96.9% identity with those of BoNT/E from C. butyricum BL 6340 and Clostridium botulinum type E, respectively. The BoNT/E-producing C. butyricum strains were divided into the following three clusters based on the results of RAPD assay, PFGE profiles of genomic DNA digested with SmaI or XhoI, and Southern blot hybridization: strains associated with infant botulism in Italy, strains associated with food-borne botulism in China, and isolates from soil specimens of the Weishan lake area in China. A DNA probe for the bont/E gene hybridized with the nondigested chromosomal DNA of all toxigenic strains tested, indicating chromosomal localization of the bont/E gene in C. butyricum. The present results suggest that BoNT/E-producing C. butyricum is clonally distributed over a vast area.

AB - Type E botulinum toxin (BoNT/E)-producing Clostridium butyricum strains isolated from botulism cases or soil specimens in Italy and China were analyzed by using nucleotide sequencing of the bont/E gene, random amplified polymorphic DNA (RAPD) assay, pulsed-field gel electrophoresis (PFGE), and Southern blot hybridization for the bont/E gene. Nucleotide sequences of the bont/E genes of 11 Chinese isolates and of the Italian strain BL 6340 were determined. The nucleotide sequences of the bont/E genes of 11 C. butyricum isolates from China were identical. The deduced amino acid sequence of BoNT/E from the Chinese isolates showed 95.0 and 96.9% identity with those of BoNT/E from C. butyricum BL 6340 and Clostridium botulinum type E, respectively. The BoNT/E-producing C. butyricum strains were divided into the following three clusters based on the results of RAPD assay, PFGE profiles of genomic DNA digested with SmaI or XhoI, and Southern blot hybridization: strains associated with infant botulism in Italy, strains associated with food-borne botulism in China, and isolates from soil specimens of the Weishan lake area in China. A DNA probe for the bont/E gene hybridized with the nondigested chromosomal DNA of all toxigenic strains tested, indicating chromosomal localization of the bont/E gene in C. butyricum. The present results suggest that BoNT/E-producing C. butyricum is clonally distributed over a vast area.

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