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Appl. Environ. Microbiol. doi:10.1128/AEM.02828-07
Copyright (c) 2008, American Society for Microbiology and/or the Listed Authors/Institutions. All Rights Reserved.

Analysis of the Neurotoxin Cluster Genes in Clostridium botulinum Strains Producing Botulinum Neurotoxin Serotype A Subtypes

Department of Bacteriology, Food Research Institute, University of Wisconsin-Madison, Madison, WI, 53706; Enteric Diseases Laboratory Branch, Centers for Disease Control and Prevention, 1600 Clifton Road, Atlanta, GA 30333

^* To whom correspondence should be addressed. Email: eajohnso{at}wisc.edu.

	Abstract

Neurotoxin cluster gene sequences and arrangements were elucidated for strains of C. botulinum encoding botulinum neurotoxin (BoNT) subtypes A3, A4, and a unique A1 producing strain (ha-/orfx+ A1). These sequences were compared to the known neurotoxin cluster sequences of C. botulinum strains that produce BoNT/A1 and BoNT/A2 and possess either a HA or an orfX cluster, respectively. The A3 and ha-/orfx+ A1 strains demonstrated a neurotoxin cluster arrangement similar to that found in A2. The A4 strain analyzed possessed two sets of neurotoxin clusters that were similar to what has been found in the A(B) strains: a HA cluster associated with the BoNT/B gene and an orfX cluster associated with the BoNT/A4 gene. The nucleotide and amino acid sequences of the neurotoxin cluster specific genes were determined for each neurotoxin cluster and compared among strains. Additionally, the ntnh gene of each strain was compared on both the nucleotide and amino acid levels. The degree of similarity of the sequences of the ntnh genes and corresponding amino acid sequences correlated with the neurotoxin cluster type to which it was associated.

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