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Applied and Environmental Microbiology, May 2008, p. 2778-2786, Vol. 74, No. 9
0099-2240/08/$08.00+0     doi:10.1128/AEM.02828-07
Copyright © 2008, American Society for Microbiology. All Rights Reserved.

Analysis of Neurotoxin Cluster Genes in Clostridium botulinum Strains Producing Botulinum Neurotoxin Serotype A Subtypes{triangledown}

Mark J. Jacobson,1 Guangyun Lin,1 Brian Raphael,2 Joanne Andreadis,2 and Eric A. Johnson1*

Department of Bacteriology, Food Research Institute, University of Wisconsin—Madison, Madison, Wisconsin 53706,1 Enteric Diseases Laboratory Branch, Centers for Disease Control and Prevention, Atlanta, Georgia 303332

Received 14 December 2007/ Accepted 21 February 2008

Neurotoxin cluster gene sequences and arrangements were elucidated for strains of Clostridium 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 hemagglutinin (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: an 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 the ntnh gene was assigned.

* Corresponding author. Mailing address: Department of Bacteriology, Food Research Institute, University of Wisconsin—Madison, 1925 Willow Drive, Madison, WI 53706. Phone: (608) 263-7944. Fax: (608) 262-9865. E-mail: eajohnso{at}wisc.edu

{triangledown} Published ahead of print on 7 March 2008.

Applied and Environmental Microbiology, May 2008, p. 2778-2786, Vol. 74, No. 9
0099-2240/08/$08.00+0     doi:10.1128/AEM.02828-07
Copyright © 2008, American Society for Microbiology. All Rights Reserved.





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