Gene Replacement in Bordetella pertussis by Transformation with Linear DNA View Full Text


Ontology type: schema:ScholarlyArticle     


Article Info

DATE

1990-11

AUTHORS

G R Zealey, S M Loosmore, R K Yacoob, S A Cockle, L J Boux, L D Miller, M H Klein

ABSTRACT

We replaced the wild-type TOX operon of Bordetella pertussis with in vitro mutated, detoxified alleles by electroporetic transformation using unmarked linear DNA. Uptake of DNA was selected by transient ampicillin resistance and two simultaneous recombination events resulted in gene-replacement at the natural locus with no integration of heterologous DNA. TOX alleles were stable without selection and recombinant strains secreted non-toxic, fully assembled, protective pertussis toxin (PT) analogues with kinetics similar to the parental vaccine strain under production-scale fermentation conditions. Strains generated in this way are suitable for the production of recombinant whole-cell or component whooping cough vaccines that require no chemical modification of PT. More... »

PAGES

1025

References to SciGraph publications

Identifiers

URI

http://scigraph.springernature.com/pub.10.1038/nbt1190-1025

DOI

http://dx.doi.org/10.1038/nbt1190-1025

DIMENSIONS

https://app.dimensions.ai/details/publication/pub.1044569101

PUBMED

https://www.ncbi.nlm.nih.gov/pubmed/1366929


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