Skip to main content

Advertisement

SpringerLink
Log in

Development of Cucumber mosaic virus as a vector modifiable for different host species to produce therapeutic proteins

  • Original Article
  • Published:
Planta Aims and scope Submit manuscript

Abstract

We have developed Cucumber mosaic virus (CMV) as a plant virus vector especially for production of pharmaceutical proteins. The CMV vector is a vector modifiable for different host plants and does not require further engineering steps. CMV contains three genomic RNA molecules (RNAs 1–3) necessary for infectivity. With this system, instead of creating different vector constructs for each plant we use, we take advantage of the formation of pseudrecombinants between two CMV isolates by simply reassembling a vector construct (RNA 2 base) and an RNA molecule containing the host determinant (mostly RNA 3). In this study, the gene for acidic fibroblast growth factor (aFGF), one of the human cytokines, was cloned under the control of the subgenomic promoter for RNA 4A of the CMV-based vector, C2-H1. Infected Nicotiana benthamiana plants produced aFGF at levels up to 5–8% of the total soluble protein. The tobacco-produced aFGF was purified, and its biological activity was confirmed. Using this system, which provides a versatile and viable strategy for the production of therapeutic proteins in plants, we also demonstrated a high level of aFGF in Glycine max (soybean) and Arabidopsis thaliana.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7

Similar content being viewed by others

Abbreviations

RT-PCR:

Reverse transcription-polymerase chain reaction

aFGF:

Acidic fibroblast growth factor

CMV:

Cucumber mosaic virus

TMV:

Tobacco mosaic virus

References

  • Arazi T, Slutsky SG, Shiboleth YM, Wang Y, Rubinstein M, Barak S, Yang J, Gal-On A (2001) Engineering zucchini yellow mosaic potyvirus as a non-pathogenic vector for expression of heterologous proteins in cucurbits. J Biotechnol 87:67–82

    Article  PubMed  CAS  Google Scholar 

  • Brigneti G, Voinnet O, Li WX, Ji LH, Ding SW, Baulcombe DC (1998) Viral pathogenicity determinants are suppressors of transgene silencing in Nicotiana benthamiana. EMBO J 17:6739–6746

    Article  PubMed  CAS  Google Scholar 

  • Burgess WH (1991) Structure–function studies of acidic fibroblast growth factor. Ann N Y Acad Sci 638:89–97

    PubMed  CAS  Google Scholar 

  • Burgess WH, Maciag T (1989) The heparin-binding (fibroblast) growth factor family of proteins. Ann Rev Biochem 58:575–606

    Article  PubMed  CAS  Google Scholar 

  • Carrère L, Tepfer M, Jacquemond M (1999) Recombinants of cucumber mosaic virus (CMV): determinants of host range and symptomatology. Arch Virol 144:365–379

    Article  PubMed  Google Scholar 

  • Charmandari E, Pincus SM, Matthews DR, Johnston A, Brook CG, Hindmarsh PC (2002) Oral hydrocortisone administration in children with classic 21-hydroxylase deficiency leads to more synchronous joint GH and cortisol secretion. J Clin Endocrinol Metab 87:2238–2244

    Article  PubMed  CAS  Google Scholar 

  • Cummins MJ, Pruitt B (1999) Low-dose oral use of human interferon-alpha in cancer patients. J Interferon Cytokine Res 19:937–941

    Article  PubMed  CAS  Google Scholar 

  • De Zoeten GA, Penswick JR, Horisberger MA, Ahl P, Schultze M, Hohn T (1989) The expression, localization, and effect of a human interferon in plants. Virology 172:213–222

    Article  PubMed  Google Scholar 

  • Ding B, Li Q, Nguyen L, Palukaitis P, Lucas WJ (1995) Cucumber mosaic virus 3a protein potentiates cell-to-cell trafficking of CMV RNA in tobacco plants. Virology 207:345–353

    Article  PubMed  CAS  Google Scholar 

  • Dirnberger D, Steinkellner H, Abdennebi L, Remy JJ, van deWiel D (2001) Secretion of biologically active glycoforms of bovine follicle stimulating hormone in plants. Eur J Biochem 268:4570–4579

    Article  PubMed  CAS  Google Scholar 

  • Edwardson JR, Christie RG (1991) Cucumoviruses. In: CRC Handbook of viruses infecting legumes, CRC Press, Boca Raton, pp 293–319

    Google Scholar 

  • Gils M, Kandzia R, Marillonnet S, Klimyuk V, Bleba Y (2005) High-yield production of authentic human growth hormone using a plant virus-based expression system. Plant Biotech J 3:613–620

    Article  CAS  Google Scholar 

  • Hong JS, Masuta C, Nakano M, Abe J, Uyeda I (2003) Adaptation of Cucumber mosaic virus soybean strains (SSVs) to cultivated and wild soybeans. Theor Appl Genet 107:49–53

    PubMed  CAS  Google Scholar 

  • James EA, Wang C, Wang Z, Reeves R, Shin JH, Magnuson NS, Lee JM (2000) Production and characterization of biologically active human GM-CSF secreted by genetically modified plant cells. Protein Expr Purif 19:131–138

    Article  PubMed  CAS  Google Scholar 

  • Kwon TH, Seo JE, Kim J, Lee JH, Jang YS, Yang MS (2003) Expression and secretion of the heterodimeric protein interleukin-12 in plant cell suspension culture. Biotechnol Bioeng 81:870–875

    Article  PubMed  CAS  Google Scholar 

  • Landgren E, Klint P, Yokote K, Claesson-Welsh L (1998) Fibroblast growth factor receptor-1 mediates chemotaxis independently of direct SH2-domain protein binding. Oncogene 17:283–291

    Article  PubMed  CAS  Google Scholar 

  • Lee HJ (2002) Protein drug oral delivery: the recent progress. Arch Pharm Res 25:572–584

    Article  PubMed  CAS  Google Scholar 

  • Lee JH, Kim NS, Kwon TH, Jang YS, Yang MS (2002) Increased production of human granulocyte-macrophage colony stimulating factor (hGM-CSF) by the addition of stabilizing polymer in plant suspension cultures. J Biotechnol 96:205–211

    Article  PubMed  CAS  Google Scholar 

  • Ma S, Huang Y, Davis A, Yin Z, Mi Q, Menassa R, Brandle JE, Jevnikar AM (2005) Production of biologically active human interleukin-4 in transgenic tobacco and potato. Plant Biotechnol J 3:309–318

    Article  PubMed  CAS  Google Scholar 

  • Magnuson NS, Linzmaier PM, Reeves R, An G, HayGlass K, Lee JM (1998) Secretion of biologically active human interleukin-2 and interleukin-4 from genetically modified tobacco cells in suspension culture. Protein Expr Purif 13:45–52

    Article  PubMed  CAS  Google Scholar 

  • Marcus PI, van der Heide L, Sekellick MJ (1999) Interferon action on avian viruses. I. Oral administration of chicken interferon-alpha ameliorates Newcastle disease. J Interferon Cytokine Res 19:881–885

    Article  PubMed  CAS  Google Scholar 

  • Marillonnet S, Giritch A, Gils M, Kandzia R, Klimyuk V, Gleba Y (2004) In planta engineering of viral RNA replicons: efficient assembly by recombination of DNA modules delivered by Agrobacterium. Proc Natl Acad Sci USA 101:6852–6857

    Article  PubMed  CAS  Google Scholar 

  • Matsumoto S, Ikura K, Ueda M, Sasaki R (1995) Characterization of a human glycoprotein (erythropoietin) produced in cultured tobacco cells. Plant Mol Biol 27:1163–1172

    Article  PubMed  CAS  Google Scholar 

  • Matsumura T, Ohya K, Itchouda N, Sugimoto C (2000) Expression of human interferon alpha and tumor necrosis factor alpha in plants. In: Toutant JP, Balázs E (eds) Molecular farming. INRA Editions, Versailles Cedex, pp 177–185

    Google Scholar 

  • Matuo Y, Nishi N, Matsumoto K, Miyazaki K, Matsumoto K, Suzuki F, Nishikawa K (1991) 3-[(3-cholamidopropyl)dimethylammonio]-1-propane sulfonate as noncytotoxic stabilizing agent for growth factors. Methods Enzymol 198:511–518

    Article  PubMed  CAS  Google Scholar 

  • Menassa R, Jevnikar A, Brandle J (2000) A contained system for the field production of plant recombinant proteins. In: Toutant JP, Balázs E (eds) Molecular farming. INRA Editions, Versailles Cedex, pp 197–205

    Google Scholar 

  • Mori M, Fujihara N, Mise K, Furusawa I (2001) Inducible high-level mRNA amplification system by viral replicase in transgenic plants. Plant J 27:79–86

    Article  PubMed  CAS  Google Scholar 

  • Nitta N, Takanami Y, Kuwata S, Kubo S (1988) Inoculation with RNAs 1 and 2 of cucumber mosaic virus induces viral RNA replicase activity in tobacco mesophyll protoplasts. J Gen Virol 69:2695–2700

    CAS  Google Scholar 

  • Ohya K, Itchoda N, Ohashi K, Onuma M, Sugimoto C, Matsumura T (2002) Expression of biologically active human tumor necrosis factor-alpha in transgenic potato plant. J Interferon Cytokine Res 22:371–378

    Article  PubMed  CAS  Google Scholar 

  • Palukaitis P, García-Arenal F (2003) Cucumoviruses. Adv Virus Res 62:241–323

    PubMed  CAS  Google Scholar 

  • Park Y, Cheong H (2002) Expression and production of recombinant human interleukin-2 in potato plants. Protein Expr Purif 25:160–165

    Article  PubMed  CAS  Google Scholar 

  • Pérez Filgueira DM, Zamorano PI, Domínguez MG, Taboga O, Del Médico Zajac MP, Puntel M, Romera SA, Morris TJ, Borca MV, Sadir AM (2003) Bovine herpes virus gD protein produced in plants using a recombinant tobacco mosaic virus (TMV) vector possesses authentic antigenicity. Vaccine 21:4201–4209

    Article  PubMed  CAS  Google Scholar 

  • Sardana RK, Alli Z, Dudani A, Tackaberry E, Panahi M, Narayanan M, Ganz P, Altosaar I (2002) Biological activity of human granulocyte-macrophage colony stimulating factor is maintained in a fusion with seed glutelin peptide. Transgenic Res 11:521–531

    Article  PubMed  CAS  Google Scholar 

  • Sawahel WA (2002) The production of transgenic potato plants expressing human alpha-interferon using lipofectin-mediated transformation. Cell Mol Biol Lett 7:19–29

    PubMed  CAS  Google Scholar 

  • Schwinghamer MW, Symons RH (1975) Fractionation of cucumber mosaic virus RNA and its translation in a wheat embryo cell-free system. Virology 63:252–262

    Article  PubMed  CAS  Google Scholar 

  • Senda M, Masuta C, Ohnishi S, Goto K, Kasai A, Sano T, Hong JS, MacFarlane S (2004) Patterning of virus-infected Glycine max seed coat is associated with suppression of endogenous silencing of chalcone synthase genes. Plant Cell 16:807–818

    Article  PubMed  CAS  Google Scholar 

  • Shin YJ, Hong SY, Kwon TH, Jang YS, Yang MS (2003) High level of expression of recombinant human granulocyte-macrophage colony stimulating factor in transgenic rice cell suspension culture. Biotechnol Bioeng 82:778–783

    Article  PubMed  CAS  Google Scholar 

  • Suzuki M, Kuwata S, Kataoka J, Masuta C, Nitta N, Takanami Y (1991) Functional analysis of deletion mutants of cucumber mosaic virus RNA3 using an in vitro transcription system. Virology 183:106–113

    Article  PubMed  CAS  Google Scholar 

  • Wagner B, Hufnagl K, Radauer C, Wagner S, Baier K, Scheiner O, Wiedermann U, Breiteneder H (2004) Expression of the B subunit of the heat-labile enterotoxin of Escherichia coli in tobacco mosaic virus-infected Nicotiana benthamiana plants and its characterization as mucosal immunogen and adjuvant. J Immunol Methods 287:203–215

    Article  PubMed  CAS  Google Scholar 

  • Wang ZD, Ueda S, Uyeda I, Yagihashi H, Sekiguchi H, Takahashi Y, Sato M, Ohya K, Sugimoto C, Matsumura T (2003) Positional effect of gene insertion on genetic stability of a clover yellow vein virus-based expression vector. J Gen Plant Pathol 69:327–334

    Article  CAS  Google Scholar 

  • Yamaguchi N, Seshimo Y, Yoshimoto E, Ahn HI, Ryu KH, Choi JK, Masuta C (2005) Genetic mapping of the compatibility between a lily isolate of Cucumber mosaic virus and a satellite RNA. J Gen Virol 86:2359–2369

    Article  PubMed  CAS  Google Scholar 

  • Zhao Y, Hammond J, Tousignant ME, Hammond RW (2000) Development and evaluation of a complementation-dependent gene delivery system based on cucumber mosaic virus. Arch Virol 145:2285–2295

    Article  PubMed  CAS  Google Scholar 

Download references

Acknowledgments

We would like to thank Dr. I. Uyeda (Hokkaido University, Sapporo, Japan) for reviewing the manuscript. This work was supported in part by Grants-in-Aid for Scientific Research (B) (no. 17380197) from the Ministry of Education, Culture, Sports, Science and Technology in Japan.

Author information

Authors and Affiliations

  1. National Institute of Advanced Industrial Science and Technology (AIST), 2-17-2-1 Tsukisamu-higashi, Toyohira-ku, Sapporo, 062-8517, Japan

    Kouki Matsuo, Jin- Sung Hong, Akira Ito & Takeshi Matsumura

  2. Hokkai Sankyo Corp., 27-4 Kitanosato, Kitahiroshima, 061-1111, Japan

    Noriko Tabayashi

  3. Graduate School of Agriculture, Hokkaido University, Kita 9, Nishi 9, Kita-ku, Sapporo, 060-8589, Japan

    Chikara Masuta

Authors
  1. Kouki Matsuo
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Jin- Sung Hong
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Noriko Tabayashi
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Akira Ito
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Chikara Masuta
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Takeshi Matsumura
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Chikara Masuta.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Matsuo, K., Hong, J.S., Tabayashi, N. et al. Development of Cucumber mosaic virus as a vector modifiable for different host species to produce therapeutic proteins. Planta 225, 277–286 (2007). https://doi.org/10.1007/s00425-006-0346-5

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00425-006-0346-5

Keywords

Search

Navigation