Identification of structural genes for Clostridium botulinum type C neurotoxin-converting phage particles

Hyun Jung Hwang, Jae Chul Lee, Yumiko Yamamoto, Mahfuzur R. Sarker, Tomofusa Tsuchiya, Keiji Oguma

Research output: Contribution to journalArticle

2 Citations (Scopus)

Abstract

The structural genes for strain C-Stockholm (c-st) phage particles, a representative type C toxin-converting phage of Clostridium botulinum, have been determined. First, by determining the N-terminal amino acid sequences of sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) bands of c-st phage particles, it became clear that four proteins, 14, 25, 32 and 42 kDa, are the products of the ORFs, cst166, cst165, cst160 and cst164, respectively, of the c-st phage genome. The Western blot analyses reacting these phage bands with an antiphage serum prepared previously indicated that the products of cst165 and cst160 are the main proteins of the phage particles. Then, six candidates for the phage structural proteins, including cst165 and cst160 gene products, were prepared as recombinant proteins. Also, the protein corresponding to the cst164 gene product was excised from SDS-PAGE gels. The antibodies against these seven proteins were prepared in rabbits, and finally, the reaction of these antibodies to the c-st phage particles was analyzed by electron microscopy. It was concluded that a sheath protein and a head protein of the c-st phage are the products of genes cst160 and cst165, respectively, and that these two proteins are conserved in the other three converting phages, but not in the nonconverting phage.

Original languageEnglish
Pages (from-to)82-89
Number of pages8
JournalFEMS Microbiology Letters
Volume270
Issue number1
DOIs
Publication statusPublished - May 2007

Fingerprint

Clostridium botulinum type C
Neurotoxins
Bacteriophages
Genes
Proteins
Sodium Dodecyl Sulfate
Polyacrylamide Gel Electrophoresis
Clostridium botulinum
Antibodies
Protein C
Recombinant Proteins
Open Reading Frames

Keywords

  • Botulinum toxin
  • Clostridium botulinum
  • Immunoelectron microscopy
  • Phage
  • Phage conversion

ASJC Scopus subject areas

  • Genetics
  • Molecular Biology
  • Applied Microbiology and Biotechnology
  • Microbiology

Cite this

Identification of structural genes for Clostridium botulinum type C neurotoxin-converting phage particles. / Hwang, Hyun Jung; Lee, Jae Chul; Yamamoto, Yumiko; Sarker, Mahfuzur R.; Tsuchiya, Tomofusa; Oguma, Keiji.

In: FEMS Microbiology Letters, Vol. 270, No. 1, 05.2007, p. 82-89.

Research output: Contribution to journalArticle

Hwang, Hyun Jung ; Lee, Jae Chul ; Yamamoto, Yumiko ; Sarker, Mahfuzur R. ; Tsuchiya, Tomofusa ; Oguma, Keiji. / Identification of structural genes for Clostridium botulinum type C neurotoxin-converting phage particles. In: FEMS Microbiology Letters. 2007 ; Vol. 270, No. 1. pp. 82-89.
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