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Journal of Bacteriology, May 2009, p. 3282-3291, Vol. 191, No. 10
0021-9193/09/$08.00+0 doi:10.1128/JB.01797-08
Copyright © 2009, American Society for Microbiology. All Rights Reserved.
Molecular Analysis of an Extrachromosomal Element Containing the C2 Toxin Gene Discovered in Clostridium botulinum Type C 
,
Yoshihiko Sakaguchi,1
Tetsuya Hayashi,2,3
Yumiko Yamamoto,1
Keisuke Nakayama,2
Kai Zhang,1
Shaobo Ma,1
Hideyuki Arimitsu,4 and
Keiji Oguma1*
Department of Bacteriology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, 2-5-1 Shikata-cho, Okayama 700-8558, Japan,1 Division of Microbiology, Department of Infectious Diseases, Faculty of Medicine, University of Miyazaki, 5200 Kiyotake, Miyazaki 899-1692, Japan,2 Division of Bioenvironmental Science, Frontier Science Research Center, University of Miyazaki, 5200 Kiyotake, Miyazaki 899-1692, Japan,3 Department of Microbiology, Fujita Health University, School of Medicine, 1-98 Dengakugakubo, Kutsukake-cho, Toyoake, Aichi 470-1192, Japan4
Received 22 December 2008/ Accepted 18 February 2009
Clostridium botulinum cultures are classified into seven types, types A to G, based on the antigenicity of the neurotoxins produced. Of these seven types, only types C and D produce C2 toxin in addition to the neurotoxin. The C2 toxin consists of two components designated C2I and C2II. The genes encoding the C2 toxin components have been cloned, and it has been stated that they might be on the cell chromosome. The present study confirmed by using pulsed-field gel electrophoresis and subsequent Southern hybridization that these genes are on a large plasmid. The complete nucleotide sequence of this plasmid was determined by using a combination of inverse PCR and primer walking. The sequence was 106,981 bp long and contained 123 potential open reading frames, including the c2I and c2II genes. The 57 products of these open reading frames had sequences similar to those of well-known proteins. It was speculated that 9 these 57 gene products were related to DNA replication, 2 were responsible for the two-component regulatory system, and 3 were 
factors. In addition, a total of 20 genes encoding proteins related to diverse processes in purine catabolism were found in two regions. In these regions, there were 9 and 11 genes rarely found in plasmids, indicating that this plasmid plays an important role in purine catabolism, as well as in C2 toxin production.
* Corresponding author. Mailing address: Department of Bacteriology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, 2-5-1 Shikata-cho, Okayama 700-8558, Japan. Phone: 81-86-235-7157. Fax: 81-86-235-7162. E-mail: kuma{at}md.okayama-u.ac.jp
Published ahead of print on 6 March 2009.
Supplemental material for this article may be found at http://jb.asm.org/.
Journal of Bacteriology, May 2009, p. 3282-3291, Vol. 191, No. 10
0021-9193/09/$08.00+0 doi:10.1128/JB.01797-08
Copyright © 2009, American Society for Microbiology. All Rights Reserved.
Copyright © 2009 by the American Society for Microbiology. For an alternate route to Journals.ASM.org, visit: http://intl-journals.asm.org | More Info»