Plant Molecular Biology

, Volume 72, Issue 1, pp 75–89

Higher accumulation of F1-V fusion recombinant protein in plants after induction of protein body formation

Authors

    • Center for Infectious Diseases and Vaccinology (CIDV)The Biodesign Institute at Arizona State University
    • Diabetes, Cardiovascular and Metabolic Diseases DivisionTranslational Genomic Research Institute (TGen)
  • Emel Topal
    • Center for Infectious Diseases and Vaccinology (CIDV)The Biodesign Institute at Arizona State University
  • Federico Martin
    • Center for Infectious Diseases and Vaccinology (CIDV)The Biodesign Institute at Arizona State University
    • Plant Molecular & Cellular Biology DepartmentUniversity of Florida
  • Guy A. Cardineau
    • Center for Infectious Diseases and Vaccinology (CIDV)The Biodesign Institute at Arizona State University
    • Departamento de Agrobiotecnología y AgronegociosTecnológico de Monterrey, Campus Monterrey
Article

DOI: 10.1007/s11103-009-9552-4

Cite this article as:
Alvarez, M.L., Topal, E., Martin, F. et al. Plant Mol Biol (2010) 72: 75. doi:10.1007/s11103-009-9552-4

Abstract

Improving foreign protein accumulation is crucial for enhancing the commercial success of plant-based production systems since product yields have a major influence on process economics. Cereal grain evolved to store large amounts of proteins in tightly organized aggregates. In maize, γ-Zein is the major storage protein synthesized by the rough endoplasmic reticulum (ER) and stored in specialized organelles called protein bodies (PB). Zera® (γ-Zein ER-accumulating domain) is the N-terminal proline-rich domain of γ-zein that is sufficient to induce the assembly of PB formation. Fusion of the Zera® domain to proteins of interest results in assembly of dense PB-like, ER-derived organelles, containing high concentration of recombinant protein. Our main goal was to increase recombinant protein accumulation in plants in order to enhance the efficiency of orally-delivered plant-made vaccines. It is well known that oral vaccination requires substantially higher doses than parental formulations. As a part of a project to develop a plant-made plague vaccine, we expressed our model antigen, the Yersinia pestis F1-V antigen fusion protein, with and without a fused Zera® domain. We demonstrated that Zera®-F1-V protein accumulation was at least 3× higher than F1-V alone when expressed in three different host plant systems: Ncotiana benthamiana, Medicago sativa (alfalfa) and Nicotiana tabacum NT1 cells. We confirmed the feasibility of using Zera® technology to induce protein body formation in non-seed tissues. Zera® expression and accumulation did not affect plant development and growth. These results confirmed the potential exploitation of Zera® technology to substantially increase the accumulation of value-added proteins in plants.

Keywords

Protein bodyZeraPlant-derived vaccinesPlant bioreactorsPlagueTransgenic plants

Abbreviations

Aa

Amino acid

ER

Endoplasmic reticulum

FD

Freeze-dried (powdered)

NT1

Nicotiana tabacum NT1 cells

PB

Protein body

SP

Signal peptide

TSP

Total soluble protein

UPR

Unfolded protein response

W.T

Wild type

Zera®

γ-Zein ER-accumulating domain

Copyright information

© Springer Science+Business Media B.V. 2009