The Use of Viral Vectors for the Production of Recombinant Proteins in Plants

  • G. P. Lomonossoff


The use of genetically engineered plant viruses to express foreign proteins or polypeptides is well established. The advantages of this method are the ease of manipulation of the small viral genome, the quick and simple procedure for inoculation of plants, and the production of high levels of recombinant proteins due to multiplication of the viruses within infected cells. This article will discuss the progress which has been made in the use of virus-based vectors and will also consider limitations to this technology.


recombinant proteins viral vectors 


  1. Ahlquist P and Janda M, 1984. cDNA cloning and in vitro transcription of the complete brome mosaic virus genome. Mol Cell Biol, 4:2876–2882Google Scholar
  2. Ahlquist P, French R, Janda M and Loesch-Fries S, 1984. Multicomponent RNA plant virus infection derived from cloned viral cDNA. Proc Natl Acad Sci USA, 81: 7066–7070PubMedCrossRefGoogle Scholar
  3. Baulcombe DC, 1999. Fast forward genetics based on virus-induced gene silencing. Curr Opin Plant Biol, 2: 109–113PubMedCrossRefGoogle Scholar
  4. Baulcombe DC, Chapman S and SantaCruz SS, 1995. Jellyfish green fluorescent protein as a reporter for virus-infections. Plant J, 7: 1045–1053PubMedCrossRefGoogle Scholar
  5. Bendahmane M, Koo M, Karrer E and Beachy RN, 1999. Display of epitopes on the surface of tobacco mosaic virus:Impact of charge and isoelectric point of the epitope on virus-host interactions. J Mol Biol, 290: 9–20PubMedCrossRefGoogle Scholar
  6. Brennan FR, Jones TD, Gilleland LB, Bellaby T, Xu F, North PC, Thompson A, Staczek J, Lin T, Johnson JE, Hamilton WDO and Gilleland HE, 1999. Pseudomonas aeroginosa outer-membrane protein F epitopes are highly immunogenic when expressed on a plant virus. Microbiology, 145: 211–220Google Scholar
  7. Burgyan J, Salanki K, Dalmay T and Russo M, 1994. Expression of homologous and heterologous viral coat protein-encoding genes using recombinant DI RNA from cymbidium ringspot tombusvirus. Gene, 138: 159–163PubMedCrossRefGoogle Scholar
  8. Chapman S, Kavanagh T and Baulcombe D, 1992. Potato virus-X as a vector for gene-expression in plants. Plant J, 2: 549–557PubMedGoogle Scholar
  9. Choi IR, Stenger DC, Morris TJ and French R, 2000. A plant virus vector for systemic expression of foreign genes in cereals. Plant J, 23: 547–555PubMedCrossRefGoogle Scholar
  10. Dalsgaard K, Uttenthal A, Jones TD, Xu F, Merryweather A, Hamilton WDO, Langeveld JPM, Boshuizen RS, Kamstrup S, Lomonossoff GP, Porta C, Vela C, Casal JI, Meloen RHand Rodgers PB, 1997. Plant-derived vaccine protects target animals against a virus disease. Nat Biotechnol, 15: 248–252PubMedCrossRefGoogle Scholar
  11. Dawson WO, Lewandowski DJ, Hilf ME, Bubrick P, Raffo AJ, Shaw JJ, Grantham GL and Desjardins PR, 1989. A tobacco mosaic virus-hybrid expresses and loses an added gene. Virology, 172: 285–292PubMedCrossRefGoogle Scholar
  12. Dolja VV, McBride HJ and Carrington JC, 1992. Tagging of plant potyvirus replication and movement by insertion of beta-glucuronidase into the viral polyprotein. Proc Natl Acad Sci USA, 89: 10208–10212PubMedCrossRefGoogle Scholar
  13. Donson J, Kearney CM, Hilf ME and Dawson WO, 1991. Systemic expression of bacterial gene by a tobacco mosaic virus-based vector. Proc Natl Acad Sci USA, 88: 7204–7208PubMedCrossRefGoogle Scholar
  14. Fernandez-Fernandez MR, Mourino M, Rivera J, Rodriguez F, Plana-Duran J and Garcia, JA, 20 01. Protection of rabbits against rabbit hemorrhagic disease virus by immunization with the VP60 ptotein expressed in plants with a potyvirus-based vector. Virology, 280: 283–291Google Scholar
  15. FitchenJ, BeachyRN, and Hein MB, 1995. Plant virus expressing hybrid coat protein with added murine epitope elicits autoantibody response. Vaccine, 13: 1051–1057CrossRefGoogle Scholar
  16. FranconiA, RoggeroP, PirazziP, AriasFJ, DesiderioA, BittiO, PashkoulovD, MatteiB, BracciL, MasengaV, MilneRG and Benvenuto E 1999. Functional expression in bacteria and plants of an scFv antibody fragment against tospoviruses. Immunotechnology, 4: 189–201CrossRefGoogle Scholar
  17. French R, Janda M and Ahlquist P,1986. Bacterial gene inserted in an engineered RNA virus–efficient expression in monocotyledonous plant-cells. Science, 231: 1294–1297Google Scholar
  18. Gopinath K, Wellink J, Porta C, Taylor KM, Lomonossoff GP and van Kammen A, 2000. Engineering cowpea mosaic virus RNA-2 into a vector to express heterologous proteins in plants. Virology, 267: 159–173PubMedCrossRefGoogle Scholar
  19. Guo HS Lopez-Moya JJ and Garcia JA, 1998. Susceptibility to recombination rearrangements of a chimeric plum pox potyvirus genome after insertion of a foreign gene. Virus Res, 57: 183–195CrossRefGoogle Scholar
  20. Hamamoto H, Sugiyama Y, Nakagawa N, Hashida E, Matsunaga Y, Takemoto S, Watanabe Y, and Okada Y, 1993. A new tobacco mosaic virus vector and its use for the systemic production of angiotensin-I-converting enzyme inhibitor in transgenic tobacco and tomato. Bio/Technology, 11: 930–932PubMedCrossRefGoogle Scholar
  21. Haynes JR, Cunningham J, von Seefried A, Lennick M, Garvin RT and Shen S-H, 1986. Development of a genetically engineered, candidate polio vaccine employing the self-assembling properties of the tobacco mosaic virus coat protein. Bio/Technology, 4: 637–641CrossRefGoogle Scholar
  22. Hendy S, Chen ZC, Barker H, SantaCruz S, Chapman S, Torrance L, Cockburn W and Whitelam, GC, 1999. Rapid production of single-chain Fv fragments in plants using a potato virus X episomal vector. J Immunol Methods, 231: 137–146PubMedCrossRefGoogle Scholar
  23. Hull R, 1978. The possible use of plant viral DNAs in genetic manipulation in plants. Trends Biochem Sci, 3: 254–256CrossRefGoogle Scholar
  24. Joelson T, Akerblom L, Oxelfelt P, Strandberg B, Tomenius K and Morris TJ, 1997. Presentation of a foreign peptide on the surface of tomato bushy stunt virus. J Gen Virol, 78: 1213–1217PubMedGoogle Scholar
  25. Kearney CM, Donson J, Jones GE and Dawson WO, 1993. Low level of genetic drift in foreign sequences replicating in a RNA virus in plants. Virology, 192: 11–17PubMedCrossRefGoogle Scholar
  26. Koo M, Bendahmane M, Lettieri GA, Paoletti AD, Lane TE, Hitchen JH, Buchmeier MJ and Beachy RN, 1999. Protective immunity against murine hepatitis virus ( MHV) induced by int.anasal or subcutaneous administration of hybrids of tobacco mosaic virus that carries an MHV epitope. Proc Nati Acad Sci USA, 96: 7774–7779Google Scholar
  27. Krebitz M, Wiedermann U, Essl D, Steinkellner H, Wagner B, Turpen TH, Ebner C, Scheirer O and Breiteneder H, 2000. Rapid production of the major birch pollen allergen Betvl in Nicotiana benthamiana plants and its immunological in vitro and in vivo characterization. FASEB J, 14: (10) 1279–1288Google Scholar
  28. Kumagai MH, Turpen TH, Weinzettl N, Della-Cioppa G, Turpen AM, Donson J, Hilf ME, Grantham GL, Dawson WO, Chow TP, Piatak M and Grill, LK, 1993. Rapid, high-level expression of biologically-active alpha-trichosanthin in transfected plants by an RNA viral vector. Proc Natl Acad Sci USA, 90: 427–430PubMedCrossRefGoogle Scholar
  29. Kumagai MH, Donson J, Della-Cioppa G and Grill LK, 2000. Rapid, high-level expression of glycosylated rice alpha-amylase in transfected plants by an RNA viral vector. Gene, 245: 169–174PubMedCrossRefGoogle Scholar
  30. Lewandowski DL and Dawson WO, 1998. Deletion of internal sequences results in tobacco mosaic virus defective RNAs that accumulate to high levels without interfering with replication of the helper virus. Virology, 251: 427–437PubMedCrossRefGoogle Scholar
  31. Lin NS, Lee YS, Lin BY, Lee CW and Hsu, YH, 1996. The open reading frame of bamboo mosaic potexvirus satellite RNA is not essential for its replication and can be replaced with a bacterial gene. Proc Natl Acad Sci USA, 93: 3138–3142PubMedCrossRefGoogle Scholar
  32. Lin T, Porta C, Lomonossoff GP and Johnson JE, 1996. Structure-based design of peptide presentation on a viral surface: The crystal structure of a plant/animal virus chimaera at 2.8A resolution. Folding and Design, 1: 179–187PubMedCrossRefGoogle Scholar
  33. Lomonossoff GP and Hamilton WDO, 1999. Cowpea mosaic virus-based vaccines. In Current topics in microbiology and immunology. Eds. Hammond J, McGarvey P and Yusibov V, Springer, Berlin, pp 177–189Google Scholar
  34. Lomonossoff GP and Johnson JE, 1996. Use of macromolecular assemblies as expression systems for peptides and synthetic vaccines. Curr Opin Struct Biol, 6: 176–182PubMedCrossRefGoogle Scholar
  35. MacFarlane SA and Popovich AH, 2000. Efficient expression of foreign proteins in roots from tobravirus vectors. Virology, 267: 29–35PubMedCrossRefGoogle Scholar
  36. Masuta C, Yamana T, Tacahashi Y, Uyeda I, Sato M, Ueda S and Matsumura T, 2000. Development of clover yellow vein virus as an efficient, stable gene-expression system for legume species. Plant J, 23: 539–546PubMedCrossRefGoogle Scholar
  37. McCormick AA, Kumagai MH, Hanley K, Turpen TH, Hakim I, Grill LK, Tuse D, Levy S and Levy R, 1999. Rapid production of specific vaccines for lymphoma by expression of the tumor-derived single-chain Fv epitopes in tobacco plants. Proc Natl Acad Sci USA, 96: 703708Google Scholar
  38. McLain L, Porta C, Lomonossoff GP, Durrani Z and Dimmock NJ, 1995. Human immunodeficiency virus type 1 neutralizing antibodies raised to a gp41 peptide expressed on the surface of a plant virus. AIDS Res Hum Retroviruses, 11: 327–334PubMedCrossRefGoogle Scholar
  39. Modelska A, Dietzschold B, Sleysh N, Fu ZF, Steplewski K, Hooper DC, Koprowski H and Yusibov V, 1998. Immunization against rabies with plant-derived antigen. Proc Natl Acad Sci USA, 95: 2481–2485PubMedCrossRefGoogle Scholar
  40. Mori M, Zhang GH, Kaido M, Okuno T and Furusawa I, 1993. Efficient production of human gamma-interferon in tobacco protoplasts by genetically-engineered brome mosaic-virus RNAs. J Gen Virol, 74: 1255–1260PubMedCrossRefGoogle Scholar
  41. O’Brien GJ, Bryant CJ, Voogd C, Greenberg HB, Gardner RC and Bellamy AR, 2000. Rotavirus VP6 expressed by PVX vectors in Nicotiana benthamiana coats PVX rods and also assembles into virus-like particles. Virology, 270: 444–453PubMedCrossRefGoogle Scholar
  42. Porta C and Lomonossoff GP, 1996. Use of viral replicons for the expression of genes in plants. Mol Biotechnol, 5: 209–211PubMedCrossRefGoogle Scholar
  43. Porta C and Lomonossoff GP, 1998. Scope for using plant viruses to present epitopes from animal pathogens. Reviews in Medical Virology, 8: 25–41PubMedCrossRefGoogle Scholar
  44. Porta C and Lomonossoff GP, 2001. Viruses as vectors for the expression of foreign sequences in Plants. Biotechnol Genet Eng Rev, 19:(in press)Google Scholar
  45. Porta C, Spall VE, Loveland J, Johnson JE, Barker PJ, and Lomonossoff GP, 1994. Development of cowpea mosaic virus as a high-yielding system for the presentation of foreign peptides. Virology, 202: 949–955Google Scholar
  46. Saitoh H, Kiba A, Nishihara M, Yamamura S, Suzuki K and Terauchi R, 2001. Production of antimicrobiol defensin in Nicotiana benthamiana with a potato virus X vector. Mol Plant-Microbe Interact, 14: 111–115PubMedCrossRefGoogle Scholar
  47. SantaCruz S, Chapman S, Roberts AG, Roberts IM, Prior DAM and Oparka KJ, 1996. Assembly and movement of a plant virus carrying a green fluorescent protein overcoat. Proc Natl Acad Sci USA, 93: 6286–6290CrossRefGoogle Scholar
  48. Scholthof HB, Moeris TJ and Jackson AO, 1993. The capsid protein gene of tomato bushy stunt virus is dispensable for systemic movement and can be replaced for localized expression of foreign genes. Mol Plant-Microbe Interact, 6: 309–322CrossRefGoogle Scholar
  49. Scholthof HB, 1999. Rapid delivery of foreign genes into plants by direct rub-inoculation with intact plasmid DNA of a tomato bushy stunt virus gene vector. J Virology, 73: 7823–7829PubMedGoogle Scholar
  50. Siegel A, 1985. Plant-virus-based vectors for gene transfer may be of considerable use despite a presumed high error frequency during RNA synthesis. Plant Mol Biol, 4: 327–329CrossRefGoogle Scholar
  51. Sjölander S, Hansen J-ES, Lövgren-Bengtsson Akerblom L and Morein B, 1996. Induction of homologous virus neutralizing antibodies in guinea pigs immunized with two human immunodeficiency virus type I glycoprotein gp 120-iscom preparations. A comparison with other adjuvant systems. Vaccine, 14: 344–352Google Scholar
  52. Smolenska L, Roberts IM, Leannonth D, Porter AJ, Harris WJ, Wilson TMA and SantaCruz S, 1998. Production of a functional single chain antibody attached to the surface of a plant virus. FEBS Lett, 441: 379–382PubMedCrossRefGoogle Scholar
  53. Sugiyama Y, I-Iamamoto H, Takemoto S, Watanabe Y and Okada Y, 1995. Systemic production of foreign peptides on the particle surface of tobacco mosaic virus. FEBS Lett, 359: 247–250PubMedCrossRefGoogle Scholar
  54. Szeto WW, Hamer DH, Carlson PS and Thomas CA, 1977. Cloning of cauliflower mosaic virus (CaMV) DNA in Escherichia coli. Science, 196: 210–212PubMedCrossRefGoogle Scholar
  55. Takamatsu N, Ishikawa M, Meshi T and Okada Y, 1987. Expression of bacterial chloramphenicol acetyltransferase gene in tobacco plants mediated by TMV-RNA. EMBO, 6: 307–311Google Scholar
  56. Takamatsu N, Watanabe Y, Yanagi H, Meshi T, Shiba T, Okada Y, 1990. Production of enkephalin in tobacco protoplasts using tobacco mosaic virus RNA vector. FEBS Lett, 269: 73–76PubMedCrossRefGoogle Scholar
  57. Taylor KM, Lin T, Porta C, Mosser AG, Giesing HA, Lomonossoff GP and Johnson JE, 2000. Influence of three-dimensional structure on the immunogenicity of a peptide expressed on the surface of a plant virus. J Mol Recognit, 13: 71–82PubMedCrossRefGoogle Scholar
  58. Taylor KM, Porta C, Lin T, Johnson JE, Barker PJ and Lomonossoff GP, 1999. Position-dependent processing of peptides presented on the surface of cowpea mosaic virus. Biol Chem, 380: 387–392PubMedCrossRefGoogle Scholar
  59. Turpen TH, Reinl SJ, Charoenvit Y, Hoffman SL, Fallarme V and Grill LK, 1995. Malarial epitopes expressed on the surface of recombinant tobacco mosaic virus. Bio/Technology, 13: 53–57PubMedCrossRefGoogle Scholar
  60. Usha R, Rohll JB, Spall VE, Shanks M, Maule AJ, Johnson JE, and Lomonossoff GP, 1993. Expression of an animal virus antigenic site on the surface of a plant virus particle. Virology, 197: 366–374PubMedCrossRefGoogle Scholar
  61. van V loten-Doting L, Bol JF and Cornelissen B, 1985. Plant virus-based vectors for gene transfer will be of limited use because of the high error frequency during viral RNA synthesis. Plant Mol Biol, 4: 323–326CrossRefGoogle Scholar
  62. Verch T, Yusibov V and Koprowski H, 1998. Expression and assembly of a full-length monoclonal antibody in plants using a plant virus vector. J Immunol Methods, 220: 69–75PubMedCrossRefGoogle Scholar
  63. Verver J, Weilink J, Van Lent J, Gopinath K and van Kammen A, 1998. Studies on the movement of cowpea mosaic virus using the jellyfish green fluorescent protein. Virology, 242: 22–27PubMedCrossRefGoogle Scholar
  64. Yusibov V, Modelska A, Steplewski K, Agadjanyan M, Weiner D, Hooper DC and Koprowski H, 1997. Antigens produced in plants by infection with chimeric plant viruses immunize against rabies virus and HIV-1. Proc Natl Acad Sci, USA, 94: 5784–5788Google Scholar
  65. Zhang GC, Leung C, Murdin L, Rovinski B and White KA, 2000. In planta expression of HIV-1 p24 protein using an RNA plant virus-based expression vector. Molec Biotechnol, 14:99–107Google Scholar
  66. Ziegler A, Cowan GH, Torrance L, Ross HA and Davies HV, 2000. Facile assessment of cDNA constructs for expression of functional antibodies in plants using the potato virus X vector. Mol Breed, 6: 327–335CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media Dordrecht 2002

Authors and Affiliations

  • G. P. Lomonossoff
    • 1
  1. 1.John Innes CentreNorwichUK

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