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Virus-Derived Vectors for the Expression of Multiple Proteins in Plants

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Recombinant Proteins from Plants

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1385))

Abstract

This chapter constitutes a practical guide to using the “pEAQ” vector series for transient or stable expression of one or more protein(s) in Nicotiana benthamiana plants. The pEAQ vectors are a series of small binary vectors designed for controlled expression of multiple proteins in plants. To achieve high levels of expression, an expression system based on translational enhancement by the untranslated regions of RNA-2 from cowpea mosaic virus (CPMV), named CPMV-HT, is used. The expression vector pEAQ-HT combines the user-friendly pEAQ plasmid with CPMV-HT to provide a system for high-level expression of proteins in plants.

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References

  1. Giritch A, Marillonnet S, Engler C, van Eldik G, Botterman J, Klimyuk V, Gleba Y (2006) Rapid high-yield expression of full-size IgG antibodies in plants coinfected with noncompeting viral vectors. Proc Natl Acad Sci U S A 103(40):14701–14706

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  2. Sainsbury F, Lavoie PO, D'Aoust MA, Vezina LP, Lomonossoff GP (2008) Expression of multiple proteins using full-length and deleted versions of cowpea mosaic virus RNA-2. Plant Biotechnol J 6(1):82–92

    CAS  PubMed  Google Scholar 

  3. Sainsbury F, Lomonossoff GP (2008) Extremely high-level and rapid transient protein production in plants without the use of viral replication. Plant Physiol 148(3):1212–1218

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  4. Sainsbury F, Thuenemann EC, Lomonossoff GP (2009) pEAQ: versatile expression vectors for easy and quick transient expression of heterologous proteins in plants. Plant Biotechnol J 7(7):682–693

    Article  CAS  PubMed  Google Scholar 

  5. Voinnet O, Rivas S, Mestre P, Baulcombe D (2003) An enhanced transient expression system in plants based on suppression of gene silencing by the p19 protein of tomato bushy stunt virus. Plant J 33(5):949–956

    Article  CAS  PubMed  Google Scholar 

  6. Sainsbury F, Saxena P, Geisler K, Osbourn A, Lomonossoff GP (2012) Using a virus-derived system to manipulate plant natural product biosynthetic pathways. Methods Enzymol 517:185–202

    Article  CAS  PubMed  Google Scholar 

  7. Thuenemann EC, Lenzi P, Love AJ, Taliansky M, Bécares M, Zuñiga S, Enjuanes L, Zahmanova GG, Minkov IN, Matić S, Noris E, Meyers A, Hattingh A, Rybicki EP, Kiselev OI, Ravin NV, Eldarov MA, Skriabin KG, Lomonossoff GP (2013) The use of transient expression systems for the rapid production of virus-like particles in plants. Curr Pharm Des 19(31):5564–5573

    Article  CAS  PubMed  Google Scholar 

  8. Vardakou M, Sainsbury F, Rigby N, Mulholland F, Lomonossoff GP (2012) Expression of active recombinant human gastric lipase in Nicotiana benthamiana using the CPMV-HT transient expression system. Protein Expr Purif 81(1):69–74

    Article  CAS  PubMed  Google Scholar 

  9. Sainsbury F, Sack M, Stadlmann J, Quendler H, Fischer R, Lomonossoff GP (2010) Rapid transient production in plants by replicating and non-replicating vectors yields high quality functional anti-HIV antibody. PLoS One 5(11):e13976

    Article  PubMed  PubMed Central  Google Scholar 

  10. Thuenemann EC, Meyers AE, Verwey J, Rybicki EP, Lomonossoff GP (2013) A method for rapid production of heteromultimeric protein complexes in plants: assembly of protective bluetongue virus-like particles. Plant Biotechnol J 11(7):839–846

    Article  CAS  PubMed  Google Scholar 

  11. Matic S, Rinaldi R, Masenga V, Noris E (2011) Efficient production of chimeric human papillomavirus 16 L1 protein bearing the M2e influenza epitope in Nicotiana benthamiana plants. BMC Biotechnol 11:106

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  12. Peyret H, Lomonossoff GP (2013) The pEAQ vector series: the easy and quick way to produce recombinant proteins in plants. Plant Mol Biol 83(1–2):51–58

    Article  CAS  PubMed  Google Scholar 

  13. Montague NP, Thuenemann EC, Saxena P, Saunders K, Lenzi P, Lomonossoff GP (2011) Recent advances of cowpea mosaic virus-based particle technology. Hum Vaccin 7(3):|383–390

    Article  CAS  PubMed  Google Scholar 

  14. Hoekema A, Hirsch PR, Hooykaas PJJ, Schilperoort RA (1983) Binary vector strategy based on separation of vir- and T-region of the Agrobacterium tumefaciens Ti-plasmid. Nature 303:179–180

    Article  CAS  Google Scholar 

  15. Meshcheriakova YA, Saxena P, Lomonossoff GP (2014) Fine-tuning levels of heterologous gene expression in plants by orthogonal variation of the untranslated regions of a nonreplicating transient expression system. Plant Biotechnol J 12(6):718–727

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  16. Saxena P, Hsieh YC, Alvarado VY, Sainsbury F, Saunders K, Lomonossoff GP, Scholthof HB (2011) Improved foreign gene expression in plants using a virus-encoded suppressor of RNA silencing modified to be developmentally harmless. Plant Biotechnol J 9(6):703–712

    Article  CAS  PubMed  Google Scholar 

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Acknowledgement

This work was supported by grant BB/J004561/1 from the Biotechnology and Biological Research Council (BBSRC) UK and the John Innes Foundation.

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Correspondence to George P. Lomonossoff .

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Saxena, P., Thuenemann, E.C., Sainsbury, F., Lomonossoff, G.P. (2016). Virus-Derived Vectors for the Expression of Multiple Proteins in Plants. In: MacDonald, J., Kolotilin, I., Menassa, R. (eds) Recombinant Proteins from Plants. Methods in Molecular Biology, vol 1385. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-3289-4_3

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  • DOI: https://doi.org/10.1007/978-1-4939-3289-4_3

  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-3288-7

  • Online ISBN: 978-1-4939-3289-4

  • eBook Packages: Springer Protocols

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