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

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

Research output: Contribution to journalArticle

41 Citations (Scopus)

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 - 1991
Externally publishedYes

Fingerprint

Brachyspira hyodysenteriae
Hemolysin Proteins
Organism Cloning
Genes
Escherichia coli
Dysentery
Spirochaetales
Osmotic Pressure
Cytotoxins
Eukaryotic Cells
Southern Blotting
Edetic Acid
Open Reading Frames
Libraries
Agar
Sequence Analysis
Swine
Cell Line
DNA

ASJC Scopus subject areas

  • Immunology

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.

Cloning and expression of a Serpula (Treponema) hyodysenteriae hemolysin gene. / Muir, S.; Koopman, M. B H; Libby, S. J.; Joens, L. A.; Heffron, Fred; Kusters, J. G.

In: Infection and Immunity, Vol. 60, No. 2, 1991, p. 529-535.

Research output: Contribution to journalArticle

Muir, S, Koopman, MBH, Libby, SJ, Joens, LA, Heffron, F & Kusters, JG 1991, 'Cloning and expression of a Serpula (Treponema) hyodysenteriae hemolysin gene', Infection and Immunity, vol. 60, no. 2, pp. 529-535.
Muir S, Koopman MBH, Libby SJ, Joens LA, Heffron F, Kusters JG. Cloning and expression of a Serpula (Treponema) hyodysenteriae hemolysin gene. Infection and Immunity. 1991;60(2):529-535.
Muir, S. ; Koopman, M. B H ; Libby, S. J. ; Joens, L. A. ; Heffron, Fred ; Kusters, J. G. / Cloning and expression of a Serpula (Treponema) hyodysenteriae hemolysin gene. In: Infection and Immunity. 1991 ; Vol. 60, No. 2. pp. 529-535.
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