Direct Expression of Bordetelia pertussis Toxin Subunits to High Levels in Escherichia coli
Author: ["W. Neal Burnette","Vernon L. Mar","Witold Cieplak","Charles F. Morris","Kaarel T. Kaljot","Kevin S. Marchitto","Raj K. Sachdev","Camille Locht","Jerry M. Keith"]
Publication: Bio/Technology
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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.
Cite this article
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
