Skip to main content
Journal of Virology logoLink to Journal of Virology
. 1990 Dec;64(12):5695–5700. doi: 10.1128/jvi.64.12.5695-5700.1990

Assembly of double-shelled, viruslike particles of bluetongue virus by the simultaneous expression of four structural proteins.

T J French 1, J J Marshall 1, P Roy 1
PMCID: PMC248707  PMID: 2173762

Abstract

Bluetongue is a disease of ruminants. The etiologic agent is bluetongue virus (BTV), a gnat-transmitted member of the Orbivirus genus of the Reoviridae. The virus has a genome of 10 double-stranded RNA species L1 to L3, M4 to M6, S7 to S10). The L2 and M5 genes of BTV which encode the outer capsid proteins VP2 and VP5, respectively, were inserted into a recombinant baculovirus downstream of duplicated copies of the baculovirus polyhedrin promoter. Insect cells coinfected with this virus plus a recombinant baculovirus expressing the two major core proteins VP3 and VP7 of BTV (T.J. French and P. Roy, J. Virol. 64:1530-1536, 1990) synthesized noninfectious, double-shelled, viruslike particles. When purified, these particles were found to have the same size and appearance as authentic BTV virions and exhibited high levels of hemagglutination activity. Antibodies raised to the expressed particles contained high titers of neutralizing activity against the homologous BTV serotype. The assembly of these bluetongue viruslike particles after the simultaneous expression of four separate proteins is indicative of the potential of this technology for the production of a new generation of viral vaccines and for the study of complex, multiprotein structures.

Full text

PDF
5700

Images in this article

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Cowley J. A., Gorman B. M. Genetic reassortants for identification of the genome segment coding for the bluetongue virus hemagglutinin. J Virol. 1987 Jul;61(7):2304–2306. doi: 10.1128/jvi.61.7.2304-2306.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Emery V. C., Bishop D. H. The development of multiple expression vectors for high level synthesis of eukaryotic proteins: expression of LCMV-N and AcNPV polyhedrin protein by a recombinant baculovirus. Protein Eng. 1987 Aug-Sep;1(4):359–366. doi: 10.1093/protein/1.4.359. [DOI] [PubMed] [Google Scholar]
  3. French T. J., Roy P. Synthesis of bluetongue virus (BTV) corelike particles by a recombinant baculovirus expressing the two major structural core proteins of BTV. J Virol. 1990 Apr;64(4):1530–1536. doi: 10.1128/jvi.64.4.1530-1536.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Huismans H., Erasmus B. J. Identification of the serotype-specific and group-specific antigens of bluetongue virus. Onderstepoort J Vet Res. 1981 Jun;48(2):51–58. [PubMed] [Google Scholar]
  5. Huismans H., van der Walt N. T., Cloete M., Erasmus B. J. Isolation of a capsid protein of bluetongue virus that induces a protective immune response in sheep. Virology. 1987 Mar;157(1):172–179. doi: 10.1016/0042-6822(87)90326-6. [DOI] [PubMed] [Google Scholar]
  6. Inumaru S., Roy P. Production and characterization of the neutralization antigen VP2 of bluetongue virus serotype 10 using a baculovirus expression vector. Virology. 1987 Apr;157(2):472–479. doi: 10.1016/0042-6822(87)90289-3. [DOI] [PubMed] [Google Scholar]
  7. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  8. Marshall J. J., Roy P. High level expression of the two outer capsid proteins of bluetongue virus serotype 10: their relationship with the neutralization of virus infection. Virus Res. 1990 Mar;15(3):189–195. doi: 10.1016/0168-1702(90)90027-9. [DOI] [PubMed] [Google Scholar]
  9. Mertens P. P., Burroughs J. N., Anderson J. Purification and properties of virus particles, infectious subviral particles, and cores of bluetongue virus serotypes 1 and 4. Virology. 1987 Apr;157(2):375–386. doi: 10.1016/0042-6822(87)90280-7. [DOI] [PubMed] [Google Scholar]
  10. Purdy M. A., Ghiasi H., Rao C. D., Roy P. Complete sequence of bluetongue virus L2 RNA that codes for the antigen recognized by neutralizing antibodies. J Virol. 1985 Sep;55(3):826–830. doi: 10.1128/jvi.55.3.826-830.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Purdy M. A., Ritter G. D., Roy P. Nucleotide sequence of cDNA clones encoding the outer capsid protein, VP5, of bluetongue virus serotype 10. J Gen Virol. 1986 May;67(Pt 5):957–962. doi: 10.1099/0022-1317-67-5-957. [DOI] [PubMed] [Google Scholar]
  12. Takehara K., Ireland D., Bishop D. H. Co-expression of the hepatitis B surface and core antigens using baculovirus multiple expression vectors. J Gen Virol. 1988 Nov;69(Pt 11):2763–2777. doi: 10.1099/0022-1317-69-11-2763. [DOI] [PubMed] [Google Scholar]
  13. Urakawa T., Ferguson M., Minor P. D., Cooper J., Sullivan M., Almond J. W., Bishop D. H. Synthesis of immunogenic, but non-infectious, poliovirus particles in insect cells by a baculovirus expression vector. J Gen Virol. 1989 Jun;70(Pt 6):1453–1463. doi: 10.1099/0022-1317-70-6-1453. [DOI] [PubMed] [Google Scholar]
  14. van der Walt N. T. A haemagglutination and haemagglutination inhibition test for bluetongue virus. Onderstepoort J Vet Res. 1980 Jun;47(2):113–117. [PubMed] [Google Scholar]

Articles from Journal of Virology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES