Biotechnology Letters

, Volume 25, Issue 13, pp 1087–1092 | Cite as

Foot-and-mouth disease virus VP1 protein fused with cholera toxin B subunit expressed in Chlamydomonas reinhardtii chloroplast

  • Meng Sun
  • Kaixian Qian
  • Ning Su
  • Huiyun Chang
  • Jixing Liu
  • Guifang Shen
Article

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.

Chlamydomonas reinhardtii chloroplast transformation cholera toxin foot-and-mouth disease virus VP1 protein 

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. Brown F (1992) Vaccination against foot and mouth disease virus. Vaccine 10: 1022–1026.Google Scholar
  2. 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.Google Scholar
  3. 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.Google Scholar
  4. 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.Google Scholar
  5. 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.Google Scholar
  6. Fan GC, Su N, Zhang ZL, Shen GF (1999) Establishment of Chlamydomonas reinhardtii chloroplast expression. Chin. Sci. Bull. 44: 1301–1306 (in Chinese).Google Scholar
  7. 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.Google Scholar
  8. Harris E (1989) The Chlamydomonas Sourcebook: A Comprehensive Guide to Biology and Laboratory Use. San Diego: Academic Press, pp. 31–52.Google Scholar
  9. 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.Google Scholar
  10. 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.Google Scholar
  11. Maliga P (2002) Engineering the plastid genome of higher plants. Curr. Opin. Plant Biol. 5: 164–172.Google Scholar
  12. 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.Google Scholar
  13. 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.Google Scholar
  14. Suzuki JY, Bauer CE (1992) Light-independent chlorophyll biosynthesis: involvement of the chloroplast gene chlL (frxC). Plant Cell 4: 929–940.Google Scholar
  15. 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.Google Scholar

Copyright information

© Kluwer Academic Publishers 2003

Authors and Affiliations

  • Meng Sun
    • 1
  • Kaixian Qian
    • 1
  • Ning Su
    • 2
  • Huiyun Chang
    • 3
  • Jixing Liu
    • 3
  • Guifang Shen
    • 2
  1. 1.Department of Biotechnology, College of Life ScienceZhejiang UniversityHangzhouP.R. China
  2. 2.Chinese Academy of Agricultural SciencesBiotechnology Research InstituteBeijingP.R. China
  3. 3.Chinese Academy of Agricultural SciencesLanzhou Veterinary Research InstituteLanzhouP.R. China

Personalised recommendations