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Journal of Bacteriology, December 2007, p. 8643-8650, Vol. 189, No. 23
0021-9193/07/$08.00+0     doi:10.1128/JB.00784-07
Copyright © 2007, American Society for Microbiology. All Rights Reserved.

Sequencing the Botulinum Neurotoxin Gene and Related Genes in Clostridium botulinum Type E Strains Reveals orfx3 and a Novel Type E Neurotoxin Subtype{triangledown}

Ying Chen,1,2 Hannu Korkeala,1 Johannes Aarnikunnas,3 and Miia Lindström1*

Department of Food and Environmental Hygiene, Faculty of Veterinary Medicine, University of Helsinki, Finland,1 College of Chemistry and Biology, Yantai University, Yantai, People's Republic of China,2 Department of Basic Veterinary Sciences, Faculty of Veterinary Medicine, University of Helsinki, Finland3

Received 20 May 2007/ Accepted 6 September 2007

Three Clostridium botulinum type E strains were sequenced for the botulinum neurotoxin (BoNT) 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 12,908 bp, and a novel gene (partial) designated orfx3, together with the complete orfx2 gene, 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 those of previously published ones. Only minor differences (≤3%) in the nucleotide sequences 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 homologies of 81%, 67 to 76%, 78 to 79%, and 79 to 85%, 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 the subtype E6 neurotoxin differed 3 to 6% from the other subtypes, suggesting that these subtype E6 neurotoxins may possess specific antigenic or functional properties.

* Corresponding author. Mailing address: Department of Food and Environmental Hygiene, P.O. Box 66, FIN-00014, University of Helsinki, Finland. Phone: 358-9-19157107. Fax: 358-9-19157101. E-mail: miia.lindstrom{at}helsinki.fi

{triangledown} Published ahead of print on 28 September 2007.

Journal of Bacteriology, December 2007, p. 8643-8650, Vol. 189, No. 23
0021-9193/07/$08.00+0     doi:10.1128/JB.00784-07
Copyright © 2007, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Lindstrom, M., Hinderink, K., Somervuo, P., Kiviniemi, K., Nevas, M., Chen, Y., Auvinen, P., Carter, A. T., Mason, D. R., Peck, M. W., Korkeala, H. (2009). Comparative Genomic Hybridization Analysis of Two Predominant Nordic Group I (Proteolytic) Clostridium botulinum Type B Clusters. Appl. Environ. Microbiol. 75: 2643-2651 [Abstract] [Full Text]  
  • Chen, Y., Korkeala, H., Linden, J., Lindstrom, M. (2008). Quantitative Real-Time Reverse Transcription-PCR Analysis Reveals Stable and Prolonged Neurotoxin Cluster Gene Activity in a Clostridium botulinum Type E Strain at Refrigeration Temperature. Appl. Environ. Microbiol. 74: 6132-6137 [Abstract] [Full Text]  
  • Artin, I., Carter, A. T., Holst, E., Lovenklev, M., Mason, D. R., Peck, M. W., Radstrom, P. (2008). Effects of Carbon Dioxide on Neurotoxin Gene Expression in Nonproteolytic Clostridium botulinum Type E. Appl. Environ. Microbiol. 74: 2391-2397 [Abstract] [Full Text]  


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