Construction of a genetically defined Salmonella typhi Ty2 aroA, aroC mutant for the engineering of a candidate oral typhoid-tetanus vaccine.
The construction of a Salmonella typhi Ty2 strain harbouring defined deletions in both the aroA and aroC genes is described. These deletions have been fully defined at the molecular level by DNA sequencing and have been introduced in such a way that no foreign DNA remains in the S. typhi genome. This strain is attenuated in mice when given by the intraperitoneal route suspended in hog gastric mucin and is attenuated to a similar level to strains harbouring deletions in aroA or aroC alone indicating that both lesions are capable of attenuating independently. We have used this defined S. typhi aroA aroC strain to express stably a non-toxic 50 kDa fragment of tetanus toxin (fragment C) from a gene incorporated into the chromosome. This strain has the advantage of harbouring no antibiotic-resistance markers and we consider it to be a candidate bivalent oral typhoid-tetanus vaccine.
Chatfield
S N
SN
Vaccine Research Unit, Wellcome Research Labs, Beckenham, Kent, UK.
Fairweather
N
N
Charles
I
I
Pickard
D
D
Levine
M
M
Hone
D
D
Posada
M
M
Strugnell
R A
RA
Dougan
G
G
D000284:Administration, Oral; D000595:Amino Acid Sequence; D000818:Animals; D002872:Chromosome Deletion; D002874:Chromosome Mapping; D003001:Cloning, Molecular; D004251:DNA Transposable Elements / Q000235:genetics; D004269:DNA, Bacterial / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics; D015870:Gene Expression / Q000235:genetics; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D009154:Mutation / Q000235:genetics; D010446:Peptide Fragments / Q000235:genetics; D012485:Salmonella typhi / Q000235:genetics* / Q000276:immunology; D013744:Tetanus Toxin / Q000235:genetics; D013745:Tetanus Toxoid / Q000235:genetics*; D014436:Typhoid-Paratyphoid Vaccines / Q000235:genetics*
D004251:DNA Transposable Elements; D004269:DNA, Bacterial; D010446:Peptide Fragments; D013744:Tetanus Toxin; D013745:Tetanus Toxoid; D014436:Typhoid-Paratyphoid Vaccines; C040635:tetanus toxin fragment C