Abstract
A Chlamydomonas reinhardtii chloroplast expression vector, pACTBVP1, containing the fusion of the foot and mouth disease virus (FMDV) VP1 gene and the cholera toxin B subunit (CTB) gene was constructed and transfered to the chloroplast genome of C. reinhardtii by the biolistic method. The transformants were identified by PCR, Southern blot, Western blot and ELISA assays after selection on resistant medium and incubation in the dark. The CTBVP1 fusion protein was expressed in C. reinhardtii chloroplast and accounted for up to 3% of the total soluble protein. The fusion protein also retained both GM1-ganglioside binding affinity and antigenicity of the FMDV VP1 and CTB proteins. These experimental results support the possibility of using transgenic chloroplasts of green alga as a mucosal vaccine source.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Brown F (1992) Vaccination against foot and mouth disease virus. Vaccine 10: 1022–1026.
Carrillo C, Wigdorovitz A, Oliveros JC, Zamorano PI, Sadir AM, Gomez N, Salinas J, Escribano JM, Borca MV (1998) Protective immune response to foot-and-mouth disease virus with VP1 expressed in transgenic plants. J. Virol. 72: 1688–1690.
Daniell H, Lee S-B, Panchal T, Wiebe PO (2001) Expression of the native cholera toxin B subunit gene and assembly as functional oligomers in transgenic tobacco chloroplasts. J. Mol. Biol. 311: 1001–1009.
DeGray G, Rajasekaran K, Smith F, Sanford J, Daniell H (2001) Expression of an antimicrobial peptide via the chloroplast genome to control phytopathogenic bacteria and fungi. Plant Physiol. 127: 852–862.
Erickson JM (1997) Chloroplast transformation: current results and future prospects. In: Ort DR, Yocum C, eds. Oxygenic Photosynthesis: The Light Reactions. Dordrecht: Kluwer Academic Publishers, pp. 589–619.
Fan GC, Su N, Zhang ZL, Shen GF (1999) Establishment of Chlamydomonas reinhardtii chloroplast expression. Chin. Sci. Bull. 44: 1301–1306 (in Chinese).
Goldschmidt-Clermont M (1991) Transgenic expression of aminoglycoside adenine transferase in the chloroplast: a selectable marker for site-directed transformation of Chlamydomonas. Nucl. Acids Res. 19: 4083–4089.
Harris E (1989) The Chlamydomonas Sourcebook: A Comprehensive Guide to Biology and Laboratory Use. San Diego: Academic Press, pp. 31–52.
Kindle KL, Richards KL, Stem DB (1991) Engineering the chloroplast genome: techniques and capabilities for chloroplast transformation in Chlamydomonas reinhardtii. Proc. Natl. Acad. Sci. USA 88: 1721–1725.
Lipscombe M, Charies IG, Roberts M, Dougan G, Tite J, Fairweather NF (1991) Intranasal immunzation using the B subunit of the Escherichia coli heat-labile toxin fused to an epitope of the Bordetella pertussis P.69 antigen. Mol. Microbiol. 5: 1385–1392.
Maliga P (2002) Engineering the plastid genome of higher plants. Curr. Opin. Plant Biol. 5: 164–172.
Sobrino F, Saiz M, Jimenez-Clavero MA, Nunez JI, Rosas MF, Baranowski E, Ley V (2001) Foot-and-mouth disease virus: a long known virus, but a current threat. Vet. Res. 32: 1–30.
Staub JM, Garcia B, Graves J, Hajdukiewicz PT, Hunter P, Nehra N (2000) High-yield production of a human therapeutic protein in tobacco chloroplasts. Nat. Biotechnol. 18: 333–338.
Suzuki JY, Bauer CE (1992) Light-independent chlorophyll biosynthesis: involvement of the chloroplast gene chlL (frxC). Plant Cell 4: 929–940.
Wigdorovitz A, Carrillo C, Dus-Santos MJ, Trono K, Peralta A, Gomez MC, Rios RD, Franzone PM, Sadir AM, Escribano JM, Borca MV (1999) Induction of a protective antibody response to foot and mouth disease virus in mice following oral or parenteral immunization with alfalfa transgenic plants expressing the viral structural protein VP1. Virology 255: 347–353.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Sun, M., Qian, K., Su, N. et al. Foot-and-mouth disease virus VP1 protein fused with cholera toxin B subunit expressed in Chlamydomonas reinhardtii chloroplast. Biotechnology Letters 25, 1087–1092 (2003). https://doi.org/10.1023/A:1024140114505
Issue Date:
DOI: https://doi.org/10.1023/A:1024140114505