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 article

190 Citations (Scopus)

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 - Feb 15 2003
Externally publishedYes

ASJC Scopus subject areas

  • Genetics

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    Tregoning, J. S., Nixon, P., Kuroda, H., Svab, Z., Clare, S., Bowe, F., Fairweather, N., Ytterberg, J., van Wijk, K. J., Dougan, G., & 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