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Direct Expression of Bordetelia pertussis Toxin Subunits to High Levels in Escherichia coli

Bio/Technology volume 6, pages 699–706 (1988)Cite this article

Abstract

The individual subunits of Bordetella pertussis toxin were subcloned and directly expressed in E. coli. The S1 subunit was synthesized at greater than 10% of total cell protein with its natural leader peptide, and at a much-reduced level as a mature methionyl polypeptide. Recombinant S1 preparations possess NAD glyco-hydrolase and ADP-ribosyltransferase activities identical to those of native pertussis toxin. Subunits S2, S3, S4, and S5 were also individually expressed in E. coli, both as mature methionyl proteins and with their authentic signal sequences, although we were unable to detect measurable amounts of S4 expressed with its signal peptide. The ability to obtain significant amounts of non-fusion recombinant sub-units will facilitate the identification of functional and enzymatic sites within the toxin, the mapping of neutralizing and protective antigenic epitopes, and will permit their assessment as candidate sub-unit vaccines for protection against whooping cough.

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Author notes

  1. Camille Locht

    Present address: Dept. of Molecular Genetics, Smith Kline-RIT, Rixensart, Belgium

  2. Kevin S. Marchitto

    Present address: NeoRx Corporation, Seattle, WA, 98119

  3. Witold Cieplak, Kaarel T. Kaljot, Kevin S. Marchitto, Camille Locht and Jerry M. Keith: Laboratory of Pathobiology, NIAID. Rocky Mountain Laboratories. Hamilton, MT 59840.

  4. W. Neal Burnette: To whom correspondence should be addressed.

Authors and Affiliations

  1. Amgen Inc., 1900 Oak Terrace Lane, Thousand Oaks, CA, 91320

    W. Neal Burnette, Vernon L. Mar, Charles F. Morris & Raj K. Sachdev

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Burnette, W., Mar, V., Cieplak, W. et al. Direct Expression of Bordetelia pertussis Toxin Subunits to High Levels in Escherichia coli. Nat Biotechnol 6, 699–706 (1988). https://doi.org/10.1038/nbt0688-699

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