1887

Abstract

To characterize the infection cycle of the multicapsid nuclear polyhedrosis virus in cells, the time course of DNA synthesis and polyhedron production, and the onset and rate of budded virus production were investigated at three different m.o.i. (5,10 and 100). In addition, the time course of expression of three proteins (gp64, p39 and polyhedrin) representative of three temporal classes of baculovirus genes was also analysed using Western blot analysis. DNA synthesis began at 12 to 18 h post-infection (p.i.). The rate of budded virus (BV) production reached maximal levels at 24 to 36 h p.i. and continued at high levels indicating that BV production was not turned off late in infection. Polyhedra were first observed at 48 h p.i. The m.o.i. appeared to influence the magnitude but not timing of early events in the viral infection cycle (gp64 expression and DNA synthesis) and also influenced the initial levels of BV production and the percentage of cells containing occlusion bodies. The m.o.i. had little influence on the final rates of BV production and the time of detection of p39 and polyhedrin on Western blots.

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1990-12-01
2024-12-08
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References

  1. Blissard G. W., Rohrmann G. F. 1989; Location, sequence, transcriptional mapping, and temporal expression of the gp64 envelope glycoprotein gene of the Orgyia pseudotsugata multicapsid nuclear polyhedrosis virus. Virology 170:537–555
    [Google Scholar]
  2. Blissard G. W., Rohrmann G. F. 1990; Baculovirus diversity and molecular biology. Annual Review of Entomology 35:127–155
    [Google Scholar]
  3. Blissard G., Quant-Russell R., Rohrmann G. F., Beaudreau G. S. 1989; Nucleotide sequence, transcriptional mapping, and temporal expression of the gene encoding p39, a major structural protein of the multicapsid nuclear polyhedrosis virus of Orgyia pseudotsugata. Virology 168:354–362
    [Google Scholar]
  4. Chisolm G. E., Henner D. J. 1988; Multiple early transcripts and splicing of the Autographa califomica nuclear polyhedrosis virus IE-1 gene. Journal of Virology 62:3193–3200
    [Google Scholar]
  5. Erlandson M. A., Carstens E. B. 1983; Mapping early transcription products of Autographa califomica nuclear polyhedrosis virus. Virology 126:398–402
    [Google Scholar]
  6. Feinberg A. P., Vogelstein B. 1983; A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity. Analytical Biochemistry 132:6–13
    [Google Scholar]
  7. Gombart A. F., Blissard G. W., Rohrmann G. F. 1989; Characterization of the genetic organization of the HindIII M region of the multicapsid nuclear polyhedrosis virus of Orgyia pseudotsugata reveals major differences among baculoviruses. Journal of General Virology 70:1815–1828
    [Google Scholar]
  8. Guarino L. A., Summers M. D. 1986; Interspersed homologous DNA of Autographa califomica nuclear polyhedrosis virus enhances delayed early gene expression. Journal of Virology 60:215–223
    [Google Scholar]
  9. Guarino L. A., Summers M. D. 1987; Nucleotide sequence and temporal expression of a baculovirus regulatory gene. Journal of Virology 61:2091–2099
    [Google Scholar]
  10. Hohmann A. W., Faulkner P. 1983; Monoclonal antibodies to baculovirus structural proteins: determination of specificities by Western blot analysis. Virology 125:432–444
    [Google Scholar]
  11. Kafatos F. C., Jones C. W., Efstratladis A. 1979; Determination of nucleic acid sequence homologies and relative concentrations by dot hybridization procedures. Nucleic Acids Research 7:1541–1552
    [Google Scholar]
  12. Laemmli U. K. 1970; Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature; London: 227680–685
    [Google Scholar]
  13. Leisy D. J., Rohrmann G. F., Beaudreau G. S. 1984; Conservation of genome organization in two multicapsid nuclear polyhedrosis viruses. Journal of Virology 52:699–702
    [Google Scholar]
  14. McClintock J. T., Dougherty E. M., Weiner R. M. 1986; Protein synthesis in gypsy moth cells infected with a nuclear polyhedrosis virus of Lymantria dispar. Virus Research 5:307–322
    [Google Scholar]
  15. Maniatis T., Fritsch E. F., Sambrook J. 1982 Molecular Cloning: A Laboratory Manual New York: Cold Spring Harbor Laboratory;
    [Google Scholar]
  16. O’Reilly D. R., Crawford A. M., Miller L. K. 1989; A viral-encoded PCNA accelerates DNA replication. Nature; London: 337606
    [Google Scholar]
  17. Pearson M. N., Russell R. L., Rohrmann G. F., Beaudreau G. S. 1988; P39, a major baculovirus structural protein: immunocyto-chemical characterization and genetic location. Virology 167:407–413
    [Google Scholar]
  18. Quant-Russell R. L., Pearson M. N., Rohrmann G. F., Beaudreau G. S. 1987; Characterization of baculovirus plO synthesis using monoclonal antibodies. Virology 160:9–19
    [Google Scholar]
  19. Reed L. J., Muench H. 1938; A simple method of estimating fifty per cent endpoints. American Journal of Hygiene 27:493–497
    [Google Scholar]
  20. Rohrmann G. F. 1977; Characterization of N-polyhedrin of two baculovirus strains pathogenic for Orgyia pseudotsugata. Biochemistry 16:1631–1634
    [Google Scholar]
  21. Rohrmann G. F. 1986; Polyhedrin structure. Journal of General Virology 61:1499–1513
    [Google Scholar]
  22. Summers M. D., Smith G. E. 1987 A Manual of Methods for Baculovirus Vectors and Insect Cell Culture Procedures Texas Agricultural Experiment Station Bulletin 1555
    [Google Scholar]
  23. Thiem S. M., Miller L. K. 1989; Identification, sequence, and transcriptional mapping of the major capsid protein gene of the baculovirus Autographa califomica nuclear polyhedrosis virus. Journal of Virology 63:2008–2018
    [Google Scholar]
  24. Tjia S., Carstens E. B., Doerfler W. 1979; Infection of Spodoptera frugiperda cells with Autographa califomica nuclear polyhedrosis virus. II. The viral DNA and the kinetics of its replication. Virology 99:399–409
    [Google Scholar]
  25. Tomalski M. D., Wu J., Miller L. K. 1988; The location, sequence, transcription and regulation of a baculovirus DNA polymerase gene. Virology 167:591–600
    [Google Scholar]
  26. Volkman L. E., Summers M. D., Hsieh C. -H. 1976; Occluded and nonoccluded nuclear polyhedrosis virus grown in Trichoplusia ni: comparative neutralization, comparative infectivity, and in vitro growth studies. Journal of Virology 19:820–832
    [Google Scholar]
  27. Whitford M., Stewart S., Kuzio J., Faulkner P. 1989; Identification and sequence analysis of a gene encoding gp67, an abundant envelope glycoprotein of the baculovirus, Autographa califomica nuclear polyhedrosis virus. Journal of Virology 63:1393–1399
    [Google Scholar]
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