Stable expression of foreign antigens from the chromosome of Salmonella typhimurium vaccine strains.
A simple and versatile system has been developed using a new cloning vector which can serve as a vehicle for integrating DNA fragments, which direct the expression of heterologous antigens, into the aroC gene on the Salmonella chromosome. The system is based on Escherichia coli plasmid vectors which contain the DNA fragment, cloned from the chromosome of S. typhimurium C5, which encodes the aroC gene. The aroC gene was modified using synthetic oligodeoxyribonucleotides so that it contained several unique restriction sites into which DNA, directing the expression of heterologous antigens, could be cloned. DNA was integrated into the S. typhimurium chromosome at aroC by transferring the vectors into S. typhimurium polA mutants and allowing homologous recombination to occur between the cloned and chromosomal aroC genes. The vectors were used to integrate nucleotide sequences into the S. typhimurium chromosome which directed the expression of tetanus toxin fragment C and the Treponema pallidum lipoprotein. The expression of both antigens was detected by Western blotting.
Department of Molecular Biology, Wellcome Biotech, Beckenham, Kent, U.K.
R A
RA
Strugnell
D
D
Maskell
N
N
Fairweather
D
D
Pickard
A
A
Cockayne
C
C
Penn
G
G
Dougan
D000942:Antigens, Bacterial / Q000235:genetics*; D001483:Base Sequence; D015153:Blotting, Western; D002874:Chromosome Mapping; D003001:Cloning, Molecular; D015870:Gene Expression; D005822:Genetic Vectors; D008190:Lyases / Q000378:metabolism; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000378:metabolism; D019761:Phosphorus-Oxygen Lyases; D011995:Recombination, Genetic; D012486:Salmonella typhimurium / Q000235:genetics* / Q000276:immunology; D013744:Tetanus Toxin / Q000378:metabolism; D014613:Vaccines, Attenuated
D000942:Antigens, Bacterial; D010446:Peptide Fragments; D013744:Tetanus Toxin; D014613:Vaccines, Attenuated; C040635:tetanus toxin fragment C; D008190:Lyases; C019895:chorismate synthase; D019761:Phosphorus-Oxygen Lyases