Cloning and expression of a Serpula (Treponema) hyodysenteriae hemolysin gene

S. Muir, M. B.H. Koopman, S. J. Libby, L. A. Joens, F. Heffron, J. G. Kusters

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    Abstract

    Serpula (Treponema) hyodysenteriae, the etiologic agent of swine dysentery, produces a hemolysin which is thought to be an important factor in the pathogenesis of the disease. We report the cloning, sequencing, and expression of a hemolysin gene (tly) from S. hyodysenteriae B204. A pUC19 gene bank of strain B204 was constructed in the Escherichia coli K-12 strain DH5α, and hemolytic recombinants were identified by plating the library on blood agar plates. From the hemolytic recombinants, a 1.5-kb DNA fragment could be isolated that contained information necessary for the production of a hemolysin/cytotoxin in E. coli. Nucleotide sequence determination of this 1.5-kb fragment showed that it contained an open reading frame capable of encoding a 26.9-kDa protein. The recombinant hemolysin was easily released from E. coli by osmotic shock. As with the native hemolysin, the recombinant hemolysin is EDTA insensitive, thermolabile, and cytotoxic for several eukaryotic cell lines. Southern blot hybridization showed that the cloned S. hyodysenteriae hemolysin gene tly is present in all pathogenic strains of S. hyodysenteriae tested and absent in the nonpathogenic, weakly hemolytic spirochete S. innocens.

    Original languageEnglish (US)
    Pages (from-to)529-535
    Number of pages7
    JournalInfection and Immunity
    Volume60
    Issue number2
    StatePublished - Dec 1 1991

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    ASJC Scopus subject areas

    • Parasitology
    • Microbiology
    • Immunology
    • Infectious Diseases

    Cite this

    Muir, S., Koopman, M. B. H., Libby, S. J., Joens, L. A., Heffron, F., & Kusters, J. G. (1991). Cloning and expression of a Serpula (Treponema) hyodysenteriae hemolysin gene. Infection and Immunity, 60(2), 529-535.