Abstract
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.
| Original language | English |
|---|---|
| Pages (from-to) | 57-63 |
| Number of pages | 7 |
| Journal | Gene |
| Volume | 88 |
| Issue number | 1 |
| DOIs | |
| Publication status | Published - 30 Mar 1990 |
| Externally published | Yes |
Keywords
- aro genes
- homologous recombination
- integration
- Recombinant DNA
- tetanus toxin
- Treponema pallidum
Cite this
}
Stable expression of foreign antigens from the chromosome of Salmonella typhimurium vaccine strains. / Strugnell, Richard A.; Maskell, Duncan; Fairweather, Neil; Pickard, Derek; Cockayne, Alan; Penn, Charles; Dougan, Gordon.
In: Gene, Vol. 88, No. 1, 30.03.1990, p. 57-63.Research output: Contribution to journal › Article › Research › peer-review
TY - JOUR
T1 - Stable expression of foreign antigens from the chromosome of Salmonella typhimurium vaccine strains
AU - Strugnell, Richard A.
AU - Maskell, Duncan
AU - Fairweather, Neil
AU - Pickard, Derek
AU - Cockayne, Alan
AU - Penn, Charles
AU - Dougan, Gordon
PY - 1990/3/30
Y1 - 1990/3/30
N2 - 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.
AB - 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.
KW - aro genes
KW - homologous recombination
KW - integration
KW - Recombinant DNA
KW - tetanus toxin
KW - Treponema pallidum
UR - http://www.scopus.com/inward/record.url?scp=0025284625&partnerID=8YFLogxK
U2 - 10.1016/0378-1119(90)90059-Z
DO - 10.1016/0378-1119(90)90059-Z
M3 - Article
VL - 88
SP - 57
EP - 63
JO - Gene
JF - Gene
SN - 0378-1119
IS - 1
ER -