Abbreviated Journal Title
Plant Biotechnol. J.
Keywords
canine parvovinus; CTB; fusion protein; GFP; peptide vaccine; plastid; transformation; TOXIN-B-SUBUNIT; NSP4 FUSION PROTEIN; CHOLERA-TOXIN; CANINE PARVOVIRUS; EDIBLE VACCINES; GENE-EXPRESSION; TRANSLATION EFFICIENCY; SYNTHETIC; PEPTIDE; SURFACE-ANTIGEN; DISEASE VIRUS; Biotechnology & Applied Microbiology; Plant Sciences
Abstract
The 2L21 peptide, which confers protection to dogs against challenge with virulent canine parvovirus (CPV), was expressed in tobacco chloroplasts as a C-terminal translational fusion with the cholera toxin B subunit (CTB) or the green fluorescent protein (GFP). Expression of recombinant proteins was dependent on plant age. A very high-yield production was achieved in mature plants at the time of full flowering (310 mg CTB-2L21 protein per plant). Both young and senescent plants accumulated lower amounts of recombinant proteins than mature plants. This shows the importance of the time of harvest when scaling up the process. The maximum level of CTB-2L21 was 7.49 mg/g fresh weight (equivalent to 31.1% of total soluble protein, TSP) and that of GFP-2L21 was 5.96 mg/g fresh weight (equivalent to 22.6% of TSP). The 2L21 inserted epitope could be detected with a CPV-neutralizing monoclonal antibody, indicating that the epitope is correctly presented at the C-terminus of the fusion proteins. The resulting chimera CTB-2L21 protein retained pentamerization and G(M1)-ganglioside binding characteristics of the native CTB and induced antibodies able to recognize VP2 protein from CPV. To our knowledge, this is the first report of an animal vaccine epitope expression in transgenic chloroplasts. The high expression of antigens in chloroplasts would reduce the amount of plant material required for vaccination (similar to100 mg for a dose of 500 mug antigen) and would permit encapsulation of freeze-dried material or pill formation.
Journal Title
Plant Biotechnology Journal
Volume
2
Issue/Number
2
Publication Date
1-1-2004
Document Type
Article
Language
English
First Page
141
Last Page
153
WOS Identifier
ISSN
1467-7644
Recommended Citation
Molina, Andrea; Hervás-Stubbs, Sandra; Daniell, Henry; Mingo-Castel, Angel M.; and Veramendi, Jon, "High-yield expression of a viral peptide animal vaccine in transgenic tobacco chloroplasts" (2004). Faculty Bibliography 2000s. 2590.
https://stars.library.ucf.edu/facultybib2000/2590