Abstract
Recently, the use of plants for the production of recombinant proteins has been well demonstrated with promising outcomes. In this study, an efficient Nicotiana tabacum L. cv. Bright Yellow 2 (BY-2) cells system expressing the ectodomain of hemagglutinin-neuraminidase (eHN) protein from Newcastle disease virus (NDV) strain AF2240 was established. Transgenic tobacco BY-2 cell cultures expressing the immunogenic eHN protein were generated and the translation efficiency of eHN protein was enhanced using the 5′-untranslated region of Nicotiana tabacum alcohol dehydrogenase gene (NtADH 5′-UTR) under the control of strong cauliflower mosaic virus (CaMV 35S) promoter. Transgenic lines verified by real-time PCR showed high level of eHN mRNA transcripts and immunoblotting confirmed the presence of 66 kD eHN protein. The eHN protein was stably produced in an average of 0.2–0.4 % total soluble protein. Green fluorescent protein-tagged eHN protein was expressed and localized at the cytosol of BY-2 cell. All mice receiving purified eHN protein from transgenic tobacco BY-2 cells produced specific anti-NDV antibodies. We concluded that plant made eHN elicit immune response and can serve as candidate vaccine against NDV.
Abbreviations
- eHN:
-
Ectodomain of hemagglutinin-neuraminidase
- GFP:
-
Green fluorescent protein
- LS:
-
Linsmaier and Skoog
- NtADH 5′-UTR:
-
5′-Untranslated region of Nicotiana tabacum alcohol dehydrogenase gene
- NDV:
-
Newcastle disease virus
References
Berinstein A, Rovere CV, Asurmendi S, Gómez E, Zanetti F, Zabal O, Tozzini A, Grand DC, Toboga O, Calmante G, Barrios H, Hopp E, Carillo E (2005) Mucosal and systemic immunization elicited by Newcastle disease virus (NDV) transgenic plants as antigens. Vaccine 23:5583–5589
Faye L, Boulaflous A, Benchabane M, Gomord V, Michaud D (2005) Protein modifications in plant secretory pathway: current status and practical implications in molecular pharming. Vaccine 23:1770–1778
Girard L, Bastin M, Courtois D (2004) Expression of the human milk protein sCD14 in tobacco plant cell culture. Plant Cell Tissue Organ Cult 78:253–260
Gómez E, Zoth SC, Asurmendi S, Rovere CV, Berinstein A (2009) Expression of hemagglutinin-neuraminidase glycoprotein of Newcastle disease virus in agroinfiltrated Nicotiana benthamiana plants. J Biotechnol 144:337–340
Haasen D, Kohler C, Neuhaus G, Merkle T (1999) Nuclear export of proteins in plants: AtXPO1 is the export receptor for leucine-rich nuclear export signals in Arabidopsis thaliana. Plant J 20:695–705
Hahn BS, Jeon IS, Jung YJ, Kim JB, Parl JS, Ha SH, Kim KH, Kim HM, Yang JS, Kim YH (2007) Expression of hemagglutinin-neuraminidase protein of Newcastle disease virus in transgenic tobacco. Plant Biotechnol Rep 1:85–92
Ko K, Koprowski H (2005) Plant biopharming of monoclonal antibodies. Virus Res 111:93–100
Martínez CA, Giulietti AM, Rodríguez Talou J (2011) Expression of a KDEL-tagged dengue virus protein in cell suspension cultures of Nicotiana tabacum and Morinda citrifolia. Plant Cell Tissue Organ Cult 107:91–100
Matsui T, Hori M, Shizawa N, Nakayama H, Shinmyo A, Yoshida K (2006) High-efficiency secretory production of peroxidase C1a using vesicular transport engineering in transgenic tobacco. J Biosci Bioeng 102:102–109
Matsui T, Asao H, Ki M, Sawada K, Kato K (2009) Transgenic lettuce producing a candidate protein for vaccine against edema disease. Biosci Biotechnol Biochem 73:1628–1634
Nakayama H, Yoshida K, Ono H, Murooka Y, Shinmyo A (2000) Ectoine, the compatible solute of Halomonas elongata, confers hyperosmotic tolerance in cultured tobacco cells. Plant Physiol 122:1239–1246
Obembe OO, Popoola JO, Leelavathi S, Reddy SV (2010) Advances in plant molecular farming. Biotechnol Adv 29:210–222
Ong HKA, Ali AM, Omar AR, Yusoff K (2000) Cloning and expression of the HN gene from velogenic viscerotropic Newcastle disease virus strain AF2240 in Sf9 insect cells. Cytotechnology 32:243–251
Pires AS, Rosa S, Castanheira S, Fevereiro P, Abranches R (2012) Expression of a recombinant human erythropoietin in suspension cultures of Arabidopsis, tobacco and Medicago. Plant Cell Tissue Organ Cult. doi:10.1007/s11240-012-0141-x
Satoh J, Kato K, Shinmyo A (2004) The 5′-untranslated region of the tobacco alcohol dehydrogenase gene functions as an effective translational enhancer in plant. J Biosci Bioeng 98:1–8
Soria-Guerra RE, Moreno-Fierros L, Roslaes-Mendoza S (2011) Two decades of plant-based candidates vaccine: a review of the chimeric protein approaches. Plant Cell Rep 30:1367–1382
Sorrentino A, Schillberg S, Fisher R, Rao R, Porta R, Mariniello L (2005) Recombinant human tissue transglutaminase produced into tobacco suspension cell cultures is active and recognizes autoantibodies in the serum of coeliac patients. Int J Biochem Cell Biol 37:842–851
Sun QY, Ding LW, Lomomossoff GP, Sun YB, Luo M, Li CQ, Jiang L, Xu ZF (2011) Improved expression and purification of recombinant human serum albumin from transgenic tobacco suspension culture. J Biotechnol 155:164–172
Urreta I, Oyanguren I, Castanón S (2011) Tobacco as biofactory for biologically active hPL production: a human hormone with the potential applications in type-1 diabetes. Transgenic Res 20:721–733
Wong SK, Tan WS, Omar AR, Tan CS, Yusoff K (2009) Immunogenic properties of recombinant ectodomian of Newcastle disease virus hemagglutinin-neuraminidase protein expressed in Escherichia coli. Act Virol 53:35–41
Yusoff K, Tan WS (2001) Newcastle disease virus: macromolecules and opportunities. Avian Pathol 30:439–455
Acknowledgments
Authors are thankful to the members of plant biotechnology and virology laboratories in UPM for their technical support.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Lai, K.S., Yusoff, K. & Mahmood, M. Functional ectodomain of the hemagglutinin-neuraminidase protein is expressed in transgenic tobacco cells as a candidate vaccine against Newcastle disease virus. Plant Cell Tiss Organ Cult 112, 117–121 (2013). https://doi.org/10.1007/s11240-012-0214-x
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s11240-012-0214-x