Abstract
The L3 and M7 genes of bluetongue virus (BTV), which encode the two major core proteins of the virus (VP3 and VP7, respectively), were inserted into a baculovirus dual-expression transfer vector and a recombinant baculovirus expressing both foreign genes isolated following in vivo recombination with wild-type Autographa californica nuclear polyhedrosis virus DNA. Spodoptera frugiperda insect cells infected with the recombinant synthesized large amounts of BTV corelike particles. These particles have been shown to be similar to authentic BTV cores in terms of size, appearance, stoichiometric arrangement of VP3 to VP7 (ratio, 2:15), and the predominance of VP7 on the surface of the particles. In infected insect cells, the corelike particles were observed in paracrystalline arrays. The formation of these structures indicates that neither the BTV double-stranded viral RNA species nor the associated minor core proteins are necessary for assembly of cores in insect cells. Furthermore, the three BTV nonstructural proteins NS1, NS2, and NS3, are not required to assist or direct the formation of empty corelike particles from VP3 and VP7.
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Selected References
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- 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]
- Ghiasi H., Purdy M. A., Roy P. The complete sequence of bluetongue virus serotype 10 segment 3 and its predicted VP3 polypeptide compared with those of BTV serotype 17. Virus Res. 1985 Sep;3(2):181–190. doi: 10.1016/0168-1702(85)90007-3. [DOI] [PubMed] [Google Scholar]
- Huismans H., van Dijk A. A., Els H. J. Uncoating of parental bluetongue virus to core and subcore particles in infected L cells. Virology. 1987 Mar;157(1):180–188. doi: 10.1016/0042-6822(87)90327-8. [DOI] [PubMed] [Google Scholar]
- Hyatt A. D., Eaton B. T. Ultrastructural distribution of the major capsid proteins within bluetongue virus and infected cells. J Gen Virol. 1988 Apr;69(Pt 4):805–815. doi: 10.1099/0022-1317-69-4-805. [DOI] [PubMed] [Google Scholar]
- 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]
- 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]
- Miller L. K. Baculoviruses as gene expression vectors. Annu Rev Microbiol. 1988;42:177–199. doi: 10.1146/annurev.mi.42.100188.001141. [DOI] [PubMed] [Google Scholar]
- Prasad B. V., Wang G. J., Clerx J. P., Chiu W. Three-dimensional structure of rotavirus. J Mol Biol. 1988 Jan 20;199(2):269–275. doi: 10.1016/0022-2836(88)90313-0. [DOI] [PubMed] [Google Scholar]
- Purdy M., Petre J., Roy P. Cloning of the bluetongue virus L3 gene. J Virol. 1984 Sep;51(3):754–759. doi: 10.1128/jvi.51.3.754-759.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Ready K. F., Sabara M. In vitro assembly of bovine rotavirus nucleocapsid protein. Virology. 1987 Mar;157(1):189–198. doi: 10.1016/0042-6822(87)90328-x. [DOI] [PubMed] [Google Scholar]
- St Angelo C., Smith G. E., Summers M. D., Krug R. M. Two of the three influenza viral polymerase proteins expressed by using baculovirus vectors form a complex in insect cells. J Virol. 1987 Feb;61(2):361–365. doi: 10.1128/jvi.61.2.361-365.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 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]
- 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]
- Verwoerd D. W., Els H. J., De Villiers E. M., Huismans H. Structure of the bluetongue virus capsid. J Virol. 1972 Oct;10(4):783–794. doi: 10.1128/jvi.10.4.783-794.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Yu Y., Fukusho A., Ritter D. G., Roy P. Complete nucleotide sequence of the group-reactive antigen VP7 gene of bluetongue virus. Nucleic Acids Res. 1988 Feb 25;16(4):1620–1620. doi: 10.1093/nar/16.4.1620. [DOI] [PMC free article] [PubMed] [Google Scholar]