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High Level Production of Hybrid Poty virus–like Particles Carrying Repetitive Copies of Foreign Antigens in Escherichia coli


Synthesis in E. coli of native coat protein of Johnsongrass mosaic virus, and hybrid protein molecules containing foreign antigens, resulted in the intracellular formation of potyvirus-like particles (PVLPs). The foreign antigens used were an octapeptide epitope from Plasmodium falciparum and a decapeptide hormone (luetinizing hormone releasing hormone) at the N- or at both N- and C- terminal regions of the coat protein molecule, and a full length protein antigen (Sj26-glutathione S-transferase of 26 kD from Schistosoma japonicum) replacing the N- terminal 62 amino acids of the coat protein. Electron microscopy of ultrathin sections of E. coli revealed that PVLPs resulting from coat protein molecules containing peptide fusions appeared in vast arrays of parallel strands within the cytoplasm sometimes extending the length of the cell and at times the cells were strung together, with “threads” of PVLPs appearing to connect individual bacterial cells. PVLPs resulting from the fusion of the 26 kD antigen Sj26 to coat protein were shorter and wider. The physical form of the high molecular weight PVLPs enabled purification by simple size exclusion column chromatography. The Sj26-PVLPs administered to mice without adjuvant elicited antibody responses comparable to monomeric Sj26 administered with Freund's Complete Adjuvant.

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  1. Hoilings, M. and Brunt, A.A. 1981. Potyviruses, p. 731–807. In: Handbook of Plant Virus Infections: Comparative Diagnosis. E. Kurstak (Ed.). Elsevier/ North-Holland, Amsterdam.

    Google Scholar 

  2. Shukla, D.D. and Ward, C.W. 1989. Structure of potyvims coat proteins and its application in the taxonomy of the poty virus group. Advances in Virus Research 36: 273–314.

    Article  CAS  Google Scholar 

  3. Bamet, O.W. 1992. Potyvirus Taxonomy. Arch. Virol. Supplementum 5.

  4. Reichmann, J.L., Lain, S. and Garcia, J.A. 1992. Highlights and prospects of potyvirus molecular biology. J. Gen. Virol. 73: 1–16.

    Article  Google Scholar 

  5. Shukla, D.D. and Ward, C.W. 1989. Identification and classification of poty viruses on the basis of coat protein sequence data and serology. Arch. Virol. 106: 171–200.

    Article  CAS  Google Scholar 

  6. Allison, R.P., Dougherty, W.G., Parks, T.D., Willis, J., Johnston, R.E., Kelly, M.E. and Armstrong, F.B. 1985. Biochemical analysis of the capsid protein gene and capsid protein of tobacco etch virus: N-terminal amino acids are located on the virion's surface. Virology 147: 309–316.

    Article  CAS  Google Scholar 

  7. Dougherty, W.G., Willis, L. and Johnston, R.F. 1985. Topographic analysis of tobacco etch virus capsid protein epitopes. Virology 144: 66–72.

    Article  CAS  Google Scholar 

  8. Shukla, D.D., Strike, P.M., Tracy, S.L., Gough, K.H. and Ward, C.W. 1988. The N and C termini of the coat proteins of poty viruses are surface-located and the N terminus contains the major virus-specific epitopes. J. Gen. Virol. 69: 1497–1508.

    Article  CAS  Google Scholar 

  9. Shukla, D.D., Tribbick, G., Mason, T.J., Hewish, D.R., Geysen, H.M. and Ward, C.W. 1989. Localization of virus-specific and group-specific epitopes of plant poryviruses by systematic immunochemical analysis of overlapping peptide fragments. Proc. Natl. Acad. Sci. USA 86: 8192–8196.

    Article  CAS  Google Scholar 

  10. McDonald, J.G., Beveridge, T.J. and Bancroft, J.B. 1976. Self-assembly of protein from a flexuous vims. Virology 69: 327–331.

    Article  CAS  Google Scholar 

  11. McDonald, J.G. and Bancroft, J.B. 1977. Assembly studies on potato virus Y and its coat protein. J. Gen. Virol. 35: 251–263.

    Article  Google Scholar 

  12. Jagadish, M.N., Ward, C.W., Gough, K.H., Tulloch, P.A., Whittaker, L.A. and Shukla, D.D. 1991. Expression of poty virus coat protein in Escherichia coli and yeast and its assembly into virus-like particles. J. Gen. Virol. 72: 1543–1550.

    Article  CAS  Google Scholar 

  13. Jagadish, M.N. and Ward, C.W. 1993. Site-directed mutagenesis of a potyvirus coat protein and its assembly in Escherichia coli . J. Gen. Virol. 74: 893–896.

    Article  CAS  Google Scholar 

  14. Saul, A., Lord, R., Jones, G.L. and Spencer, L. 1992. Protective immunization with invariant peptides of the Plasmodium falciparum antigen MSA2. J. Immunol. 148: 208–211.

    CAS  PubMed  Google Scholar 

  15. Smythe, J.A., Coppel, R.L., Brown, G.V., Ramasamy, R., Kemp, D.J. and Anders, R.F. 1988. Identification of two integral membrane proteins of Plasmodium falciparum . Proc. Natl. Acad. Sci. USA 85: 5195–5199.

    Article  CAS  Google Scholar 

  16. Fraser, H.M. and Gunn, A. 1973. Effects of antibodies to luteinizing hormone-releasing hormone in the male rabbit and on the rat oestrus cycle. Nature 244: 160–161.

    Article  CAS  Google Scholar 

  17. Smith, D.B., Davern, K.M., Board, P.G., Tiu, W.U., Garcia, E.G. and Mitchell, G.F. 1986. Mr 26,000 antigen of Schistosoma japonicum recognized by resistant WEHI 129/J mice is a parasite glutathione S-transferase. Proc. Natl. Acad. Sci. USA 83: 8703–8707.

    Article  CAS  Google Scholar 

  18. Smith, D.B. and Johnson, K.S. 1988. Single-step purification of polypeptides expressed in Escherichia coli as fusions with glutathione S-transeferase. Gene 67: 31–40.

    Article  CAS  Google Scholar 

  19. Sambrook, J., Fritsch, E.F. and Maniatis, T. 1989. Molecular Cloning: A Laboratory Manual. Cold Spring Harbor Laboratory Press, NY.

    Google Scholar 

  20. Harlow, E. and Lane, D. 1988. Antibodies: A Laboratory Manual. Cold Spring Harbor Laboratory Press, NY.

    Google Scholar 

  21. Davern, K.M., Wright, M.D., Hermann, V.R. and Mitchell, G.F. 1991. Further characterisation of Schistosoma japonicum protein SJ26, a target antigen of an immunodiagnostic monoclonal antibody. Mol. and Bloc-hem. Parasitology 48: 67–76.

    Article  CAS  Google Scholar 

  22. Smith, D.B., Rubira, M.R., Simpson, R.J., Davern, K.M., Tiu, W.U., Board, P.G. and Mitchell, G.F. 1988. Expression of an enzymatically active parasite molecule in Escherichia coli: Schistosoma japonicum glutathione S-transferase. Mol. Biochem. Parasitol. 27: 249–256.

    Article  CAS  Google Scholar 

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Jagadish, M., Hamilton, R., Fernandez, C. et al. High Level Production of Hybrid Poty virus–like Particles Carrying Repetitive Copies of Foreign Antigens in Escherichia coli. Nat Biotechnol 11, 1166–1170 (1993).

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