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J. Bacteriol. doi:10.1128/JB.00784-07
Copyright (c) 2007, American Society for Microbiology and/or the Listed Authors/Institutions. All Rights Reserved.

Sequencing the botulinum neurotoxin and related genes in Clostridium botulinum type E strains reveals orfx3 and a novel type E neurotoxin subtype

Department of Food and Environmental Hygiene, Faculty of Veterinary Medicine, University of Helsinki, Finland; College of Chemistry and Biology, Yantai University, PR China; Department of Basic Veterinary Sciences, Faculty of Veterinary Medicine, University of Helsinki, Finland

^* To whom correspondence should be addressed. Email: miia.lindstrom{at}helsinki.fi.

	Abstract

Three Clostridium botulinum type E strains were sequenced for the botulinum neurotoxin gene cluster, and 11 type E strains, representing a wide biodiversity, were sequenced for the bont/E gene. The total length of the BoNT/E gene cluster was 12908 bp, and a novel gene (partial) denoted orfx3, together with complete orfx2, was identified in the three type E strains for the first time. Apart from orfx3, the structure and organization of the neurotoxin gene cluster of the three strains were identical to previously published ones. Only minor differences ( 3%) in the nucleotide sequence of the gene cluster components were observed among the three strains and the published BoNT/E-producing clostridia. The orfx3, orfx2, orfx1 and p47 gene sequences of the three type E strains shared 81%, 67-76%, 78-79% and 79-85% homology, respectively, with published sequences for type A1 and A2 C. botulinum. Analysis of bont/E from the 14 type E strains and 19 previously published BoNT/E-producing clostridia revealed six neurotoxin subtypes, with a new distinct subtype consisting of three Finnish isolates alone. The amino acid sequence of subtype E6 neurotoxin differed by 3-6% from the other subtypes, suggesting that these subtype E6 neurotoxins may possess specific antigenic or functional properties.

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