Expression of tetanus toxin Fragment C in tobacco chloroplasts

John S. Tregoning, Peter Nixon, Hiroshi Kuroda, Zora Svab, Simon Clare, Frances Bowe, Neil Fairweather, Jimmy Ytterberg, Klaas J. van Wijk, Gordon Dougan, Pal Maliga

Research output: Contribution to journalReview ArticleResearchpeer-review

Abstract

Fragment C (TetC) is a non-toxic 47 kDa polypeptide fragment of tetanus toxin that can be used as a subunit vaccine against tetanus. Expression of TetC in Escherichia coli and yeast was dependent on the availability of synthetic genes that were required to improve translation efficiency and stabilize the mRNA. To explore the feasibility of producing TetC in tobacco leaves, we attempted expression of both the bacterial high-AT (72.3% AT) and the synthetic higher-GC genes (52.5% AT) in tobacco chloroplasts. We report here that the bacterial high-AT mRNA is stable in tobacco chloroplasts. Significant TetC accumulation was obtained from both genes, 25 and 10% of total soluble cellular protein, respectively, proving the versatility of plastids for expression of unmodified high-AT and high-GC genes. Mucosal immunization of mice with the plastid-produced TetC induced protective levels of TetC antibodies. Thus, expression of TetC in chloroplasts provides a potential route towards the development of a safe, plant-based tetanus vaccine for nasal and oral applications.

Original languageEnglish
Pages (from-to)1174-1179
Number of pages6
JournalNucleic Acids Research
Volume31
Issue number4
DOIs
Publication statusPublished - 15 Feb 2003
Externally publishedYes

Cite this

Tregoning, J. S., Nixon, P., Kuroda, H., Svab, Z., Clare, S., Bowe, F., ... Maliga, P. (2003). Expression of tetanus toxin Fragment C in tobacco chloroplasts. Nucleic Acids Research, 31(4), 1174-1179. https://doi.org/10.1093/nar/gkg221
Tregoning, John S. ; Nixon, Peter ; Kuroda, Hiroshi ; Svab, Zora ; Clare, Simon ; Bowe, Frances ; Fairweather, Neil ; Ytterberg, Jimmy ; van Wijk, Klaas J. ; Dougan, Gordon ; Maliga, Pal. / Expression of tetanus toxin Fragment C in tobacco chloroplasts. In: Nucleic Acids Research. 2003 ; Vol. 31, No. 4. pp. 1174-1179.
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abstract = "Fragment C (TetC) is a non-toxic 47 kDa polypeptide fragment of tetanus toxin that can be used as a subunit vaccine against tetanus. Expression of TetC in Escherichia coli and yeast was dependent on the availability of synthetic genes that were required to improve translation efficiency and stabilize the mRNA. To explore the feasibility of producing TetC in tobacco leaves, we attempted expression of both the bacterial high-AT (72.3{\%} AT) and the synthetic higher-GC genes (52.5{\%} AT) in tobacco chloroplasts. We report here that the bacterial high-AT mRNA is stable in tobacco chloroplasts. Significant TetC accumulation was obtained from both genes, 25 and 10{\%} of total soluble cellular protein, respectively, proving the versatility of plastids for expression of unmodified high-AT and high-GC genes. Mucosal immunization of mice with the plastid-produced TetC induced protective levels of TetC antibodies. Thus, expression of TetC in chloroplasts provides a potential route towards the development of a safe, plant-based tetanus vaccine for nasal and oral applications.",
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Tregoning, JS, Nixon, P, Kuroda, H, Svab, Z, Clare, S, Bowe, F, Fairweather, N, Ytterberg, J, van Wijk, KJ, Dougan, G & Maliga, P 2003, 'Expression of tetanus toxin Fragment C in tobacco chloroplasts', Nucleic Acids Research, vol. 31, no. 4, pp. 1174-1179. https://doi.org/10.1093/nar/gkg221

Expression of tetanus toxin Fragment C in tobacco chloroplasts. / Tregoning, John S.; Nixon, Peter; Kuroda, Hiroshi; Svab, Zora; Clare, Simon; Bowe, Frances; Fairweather, Neil; Ytterberg, Jimmy; van Wijk, Klaas J.; Dougan, Gordon; Maliga, Pal.

In: Nucleic Acids Research, Vol. 31, No. 4, 15.02.2003, p. 1174-1179.

Research output: Contribution to journalReview ArticleResearchpeer-review

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T1 - Expression of tetanus toxin Fragment C in tobacco chloroplasts

AU - Tregoning, John S.

AU - Nixon, Peter

AU - Kuroda, Hiroshi

AU - Svab, Zora

AU - Clare, Simon

AU - Bowe, Frances

AU - Fairweather, Neil

AU - Ytterberg, Jimmy

AU - van Wijk, Klaas J.

AU - Dougan, Gordon

AU - Maliga, Pal

PY - 2003/2/15

Y1 - 2003/2/15

N2 - Fragment C (TetC) is a non-toxic 47 kDa polypeptide fragment of tetanus toxin that can be used as a subunit vaccine against tetanus. Expression of TetC in Escherichia coli and yeast was dependent on the availability of synthetic genes that were required to improve translation efficiency and stabilize the mRNA. To explore the feasibility of producing TetC in tobacco leaves, we attempted expression of both the bacterial high-AT (72.3% AT) and the synthetic higher-GC genes (52.5% AT) in tobacco chloroplasts. We report here that the bacterial high-AT mRNA is stable in tobacco chloroplasts. Significant TetC accumulation was obtained from both genes, 25 and 10% of total soluble cellular protein, respectively, proving the versatility of plastids for expression of unmodified high-AT and high-GC genes. Mucosal immunization of mice with the plastid-produced TetC induced protective levels of TetC antibodies. Thus, expression of TetC in chloroplasts provides a potential route towards the development of a safe, plant-based tetanus vaccine for nasal and oral applications.

AB - Fragment C (TetC) is a non-toxic 47 kDa polypeptide fragment of tetanus toxin that can be used as a subunit vaccine against tetanus. Expression of TetC in Escherichia coli and yeast was dependent on the availability of synthetic genes that were required to improve translation efficiency and stabilize the mRNA. To explore the feasibility of producing TetC in tobacco leaves, we attempted expression of both the bacterial high-AT (72.3% AT) and the synthetic higher-GC genes (52.5% AT) in tobacco chloroplasts. We report here that the bacterial high-AT mRNA is stable in tobacco chloroplasts. Significant TetC accumulation was obtained from both genes, 25 and 10% of total soluble cellular protein, respectively, proving the versatility of plastids for expression of unmodified high-AT and high-GC genes. Mucosal immunization of mice with the plastid-produced TetC induced protective levels of TetC antibodies. Thus, expression of TetC in chloroplasts provides a potential route towards the development of a safe, plant-based tetanus vaccine for nasal and oral applications.

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JF - Nucleic Acids Research

SN - 0305-1048

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Tregoning JS, Nixon P, Kuroda H, Svab Z, Clare S, Bowe F et al. Expression of tetanus toxin Fragment C in tobacco chloroplasts. Nucleic Acids Research. 2003 Feb 15;31(4):1174-1179. https://doi.org/10.1093/nar/gkg221