Expression of tetanus toxin Fragment C in tobacco chloroplasts

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 stabiliz...

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Bibliographic Details
Published in:Nucleic acids research 2003-02, Vol.31 (4), p.1174-1179
Main Authors: Tregoning, John S., Nixon, Peter, Kuroda, Hiroshi, Svab, Zora, Clare, Simon, Bowe, Frances, Fairweather, Neil, Ytterberg, Jimmy, Wijk, Klaas J. van, Dougan, Gordon, Maliga, Pal
Format: Article
Language:English
Subjects:
Animals
Blotting, Northern
Blotting, Western
Chloroplasts - genetics
Gene Expression
Immunoglobulin G - immunology
Immunoglobulin G - metabolism
Mice
Nicotiana - genetics
Peptide Fragments - genetics
Peptide Fragments - immunology
Peptide Fragments - metabolism
Plant Extracts - immunology
Plant Leaves - chemistry
Plants, Genetically Modified
Plasmids - genetics
RNA, Messenger - genetics
RNA, Messenger - metabolism
Tetanus Toxin - genetics
Tetanus Toxin - immunology
Tetanus Toxin - metabolism
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Summary: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.
ISSN:0305-1048
1362-4962
1362-4962
DOI:10.1093/nar/gkg221