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 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.
Tregoning
John S
JS
Waksman Institute, Rutgers University, 190 Frelinghuysen Road, Piscataway, NJ 08854-8020, USA.
Nixon
Peter
P
Kuroda
Hiroshi
H
Svab
Zora
Z
Clare
Simon
S
Bowe
Frances
F
Fairweather
Neil
N
Ytterberg
Jimmy
J
van Wijk
Klaas J
KJ
Dougan
Gordon
G
Maliga
Pal
P
D000818:Animals; D015152:Blotting, Northern; D015153:Blotting, Western; D002736:Chloroplasts / Q000235:genetics*; D015870:Gene Expression; D007074:Immunoglobulin G / Q000276:immunology / Q000378:metabolism; D051379:Mice; D010446:Peptide Fragments / Q000235:genetics* / Q000276:immunology / Q000378:metabolism; D010936:Plant Extracts / Q000276:immunology; D018515:Plant Leaves / Q000737:chemistry; D030821:Plants, Genetically Modified; D010957:Plasmids / Q000235:genetics; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D013744:Tetanus Toxin / Q000235:genetics* / Q000276:immunology / Q000378:metabolism; D014026:Nicotiana / Q000235:genetics*
D007074:Immunoglobulin G; D010446:Peptide Fragments; D010936:Plant Extracts; D012333:RNA, Messenger; D013744:Tetanus Toxin; C040635:tetanus toxin fragment C