Molecular Biotechnology

, Volume 32, Issue 2, pp 117–127

Synthesis of a ricin toxin B subunit-rotavirus VP7 fusion protein in potato

  • Nak-Won Choi
  • Mary K. Estes
  • William H. R. Langridge
Research

DOI: 10.1385/MB:32:2:117

Cite this article as:
Choi, NW., Estes, M.K. & Langridge, W.H.R. Mol Biotechnol (2006) 32: 117. doi:10.1385/MB:32:2:117

Abstract

A gene encoding the outer capsid glycoprotein (VP7) of simian rotavirus SA11, was genetically linked to the amino terminus of the ricin toxin B subunit (RTB) isolated from castor-oil plant (Ricinus communis) seeds. To assess fusion protein expression in plant cells, the VP7::RTB fussion gene was transferred into potato (Solanum tuberosum) cells by Agrobacterium tumefaciens-mediated transformation methods and transformed plants regenerated. The fusion gene was detected in transformed potato genomic DNA by polymerase chain reaction DNA amplification methods. Immunoblot analysis with anti-SA11 antiserum as the primary antibody verified the presence of VP7::RTB fusion protein in transformed potato tuber tissues. The plant-synthesized fusion protein bound RTB membrane receptors as measured by asialofetuin-enzyme-linked immunosorbent assay (ELISA). The ELISA results indicated that the VP7::RTB fusion protein was biologically active and made up approx 0.03% of total soluble transformed tuber protein. The biosynthesis of receptor binding VP7::RTB fusion protein in potato tissues demonstrates the feasibility of producing monomeric ricin toxin B subunit adjuvant-virus antigen fusion proteins in crop plants for enhanced immunity.

Index Entries

Rotavirus outer capsid protein VP7 ricin toxin B subunit biological activity edible subunit vaccine Solanum tuberosum 

Copyright information

© Humana Press Inc 2006

Authors and Affiliations

  • Nak-Won Choi
    • 1
  • Mary K. Estes
    • 2
  • William H. R. Langridge
    • 1
  1. 1.Center for Molecular Biology and Gene Therapy, Department of Biochemistry and Microbiology, School of MedicineLoma Linda UniversityLoma Linda
  2. 2.Department of Molecular Virology and MicrobiologyBaylor College of MedicineHouston

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