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Applied and Environmental Microbiology, November 2000, p. 4992-4997, Vol. 66, No. 11
Department of Bacteriology, School of
Medicine, Kanazawa University, Kanazawa
920-8640,1 Department of Veterinary
Science, College of Agriculture, Osaka Prefecture University, Sakai,
Osaka 599-8531,2 Department of
Bacteriology, Toyama Institute of Health, Toyama
939-0363,3 and Department of
Bacteriology, Okayama University Medical School, Okayama
700-8558,4 Japan
Received 19 April 2000/Accepted 28 August 2000
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.
0099-2240/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.
Genetic Analysis of Type E Botulinum
Toxin-Producing Clostridium butyricum Strains
*
Corresponding author. Mailing address: Department of
Bacteriology, School of Medicine, Kanazawa University, Kanazawa
920-8640, Japan. Phone: 81-76-265-2200. Fax: 81-76-234-4230. E-mail:
nakamura{at}med.kanazawa-u.ac.jp.
We dedicate this work to the memory of C. L. Hatheway.
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