Connaught Laboratories Limited

Aventis Pasteur Limited

Gavin Ross Zealey

Michel Henri Klein

Sheena M Loosmore

Reza Khayyam Yacoob

Stephen A Cockle

Heather A Boux

Autologous promoters are used to effect expression of gene products in Bordetella strains. Hybrid Bordetella genes are constructed comprising a Bordetella gene, particularly one encoding a Bordetella antigen, fused at an ATG codon to a native but autologous Bordetella promoter or other Bordetella strain. Genes and promoters from are preferred. , containing the hybrid gene by insertion into the chromosome of the organism by homologous recombination at specific loci, effects expression of the protein for which the Bordetella gene codes at a production rate different from that achieved for the homologous gene. Specific strains and plasmids are described.

Use of autologous promoters to express gene products in Bordetella

An expression system for expressing gene products from recombinant Bordetella strains and specific nucleic acid molecules useful in transforming Bordetella strains for such expression are described. A nucleic acid molecule may comprise a Bordetella promoter operatively coupled to a heterologous gene encoding a non-Bordetella gene product with the heterologous gene transcriptionally regulated by the Bordetella promoter. The nucleic acid molecule may further comprise a further nucleic acid molecule encoding a leader sequence for secretion of the non-Bordetella gene product. Another nucleic acid molecule may comprise a Bordetella promoter coupled to a nucleic acid sequence encoding a non-Bordetella leader sequence for secretion of a gene product, which may be a Bordetella gene product or a non-Bordetella gene product.

Expression of gene products from genetically manipulated strains of

Autologous promoters are used to effect expression of gene products in Bordetella strains. Hybrid Bordetella genes are constructed comprising a Bordetella gene, particularly one encoding a Bordetella antigen, fused at an ATG codon to a native but autologous Bordetella promoter or other Bordetella strain. Genes and promoters from B. pertussis are preferred. B. pertussis, containing the hybrid gene by insertion into the chromosome of the organism by homologous recombination at specific loci, effects expression of the protein for which the Bordetella gene codes at a production rate different from that achieved for the homologous gene. Specific strains and plasmids are described.

Use of autologous promoters to express gene products in bordetella

Protein expression levels from Bordetella strains, particularly Bordetella pertussis, are altered by genetic modification to a natural Bordetella strain whereby one or more of the natural genes, particularly including the TOX, FHA, CYA and PRN genes, is deleted from the genome of the natural strain and one or more of the natural genes or a genetic mutation thereof, particularly a genetically-detoxified TOX* gene, or a hybrid gene, is inserted into the genome of the natural strain to provide at least two copies of one or more of the natural genes or genetic mutation thereof or hybrid gene, singly or in tandem. The altered genotype Bordetella strain is useful in producing whole-cell or defined component vaccines against Bordetella, particularly whooping cough, which may be employed in combination with other vaccines.

Manipulation of gene copy number in bordetella

A new method is described for the preparation of a safe, immunogenic and efficacious vaccine for protection against the disease pertussis. In development of this vaccine, specific functional sites of pertussis toxin have been identified, and using this information, defined mutant holotoxins have been produced by site directed mutagenesis of the toxin gene. A number of these holotoxin analogues are detoxified, retain an immunodominant S1 epitope, are immunogenic and are protective in the standard pertussis vaccine potency test in mice.

Immunoprotective genetically-detoxified mutants of pertussis toxin

Genetic detoxification of pertussis toxin

Vaccine containing genetically-detoxified pertussis holotoxin

A new method is described for the preparation of a safe, immunogenic and efficacious vaccine for protection against the disease pertussis. In development of this vaccine, specific functional sites of pertussis toxin have been identified, and using this information, defined mutant holotoxins have been produced by site directed mutagenesis of the toxin gene. A number of these holotoxin analogs are detoxified, retain an immunodominant S1 epitope, are immunogenic and are protective in the standard pertussis vaccine potency test in mice.

A new method is described for the preparation of a safe, immunogenic and efficacious vaccine for protection against the disease pertussis. In development of this vaccine, specific functional sites of pertussis toxin have been identified, and using this information, defined mutant holotoxins have been produced by site directed mutagenesis of the toxin gene. A number of these toxin analogues are detoxified, retain an immunodominant S1 epitope, are immunogenic and are protective in the standard pertussis vaccine potency test in mice.

A new method is described for the preparation of a safe, immunogenic and efficacious vaccine for protection against the disease pertussis. In development of this vaccine, specific functional sites of pertussis toxin have been identified, and using this information, defined mutant holotoxins have been produced by site directed mutagenesis of the toxin gene. A number of these toxin analogues are detoxified, retain an immunodominant S1 epitope, are immunoganic and are protective in the standard pertussis vaccine potency test in mice.

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Thornhill, Canada

Gavin Ross Zealey