1887

Abstract

SUMMARY

A cotransfection method has been developed for the generation of recombinant baculoviruses. The permissive coleopteran cell-line DSIR-HA-1179 was transfected with a mixture of baculovirus DNA (strain PV505) and a transfer vector. The transfer vector, a pUC8-based plasmid, contained the polyhedrin gene from the nuclear polyhedrosis virus, flanked by baculovirus DNA from the III fragment N. baculovirus does not form plaques with DSIR-HA-1179 cells. Therefore an endpoint dilution method was used to screen for, recover and purify recombinants. There was no phenotypic character that could be used for detecting recombinants, so a dot blot assay was used to screen infected cultures for the presence of recombinants. A recombinant generated by this method contained the entire polyhedrin gene inserted at 98.03 map units from the designated start of the baculovirus physical map. No evidence of transcription or translation of the polyhedrin gene was obtained.

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1989-04-01
2024-04-19
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References

  1. Andrews M. E., Boyle D. B., Coupar B. E.H., Whitfield P. L., Both G. W., Bellamy A. R. 1987; Vaccinia virus recombinants expressing the SA11 rotavirus VP7 glycoprotein induce serotype-specific neutralizing antibodies. Journal of Virology 61:1054–1060
    [Google Scholar]
  2. Bedford G. O. 1981; Control of the rhinoceros beetle by baculovirus. Microbial Control of Pests and Plant Diseases 1970–1980409–426 Burges H. D. London: Academic Press;
    [Google Scholar]
  3. Burand J. P., Summers M. D., Smith G. E. 1980; Transfection with baculovirus DNA. Virology 101:286–290
    [Google Scholar]
  4. Carbonell L. F., Klowden M. J., Miller L. K. 1985; Baculovirus-mediated expression of bacterial genes in dipteran and mammalian cells. Journal of Virology 56:153–160
    [Google Scholar]
  5. Chirgwin J. M., Przybla A. E., Macdonald R. J., Rutter W. J. 1979; Isolation of biologically active RNA from sources enriched in ribonuclease. Biochemistry 18:5294–5299
    [Google Scholar]
  6. Crawford A. M. 1982; A Coleopteran cell line derived from Heteronychus arator (Coleoptera: Scarabaeidae). In Vitro 18:813–816
    [Google Scholar]
  7. Crawford A. M., Sheehan C. 1985; Replication of Oryctes baculovirus in cell culture: viral morphogenesis, infectivity and protein synthesis. Journal of General Virology 66:529–539
    [Google Scholar]
  8. Crawford A. M., Sheehan C. M., King P. D., Meekings J. 1985a; Oryctes baculovirus infectivity for New Zealand scarabs. Proceedings of the 4th Australasian Conference on Grassland Invertebrate Ecology224–227 Chapman. R. B., Christchurch N.Z. Caxton Press;
    [Google Scholar]
  9. Crawford A. M., Ashbridge K., Sheehan C., Faulkner P. 1985b; A physical map of the Oryctes baculovirus genome. Journal of General Virology 66:2649–2658
    [Google Scholar]
  10. Crawford A. M., Zelazny B., Alfiler A. R. 1986; Genotypic variation in geographical isolates of Oryctes baculovirus. Journal of General Virology 67:949–952
    [Google Scholar]
  11. Fung M. C., Emu K. Y. M., Weber T., Chang T. W., Chang N. T. 1988; Detection and purification of a recombinant human B lymph trophic virus (HIV-6) in the baculovirus expression system by limiting dilution and DNA dot-blot hybridisation. Journal of Virological Methods 19:33–42
    [Google Scholar]
  12. Gonnet P., Devauchelle G. 1987; Obtention par recombinaison dans le gène du polypeptide p10 d’un baculovirus exprimant le gène de résistance à la néomycine dans les cellules d’insecte. Comptes rendus de l’Acadámie des Sciences 305:III111–114
    [Google Scholar]
  13. Graham F. L., Van Der Eb A. J. 1973; A new technique for the assay of infectivity of human adenovirus 5 DNA. Virology 52:456–67
    [Google Scholar]
  14. Guilford P. J., Forster R. L. S. 1986; Detection of polyadenylated subgenomic RNAs in leaves infected with the potexvirus, daphne virus X. Journal of General Virology 67:83–90
    [Google Scholar]
  15. Hooft Van Iddekinge B. J. L., Smith G. E., Summers M. D. 1983; Nucleotide sequence of the polyhedrin gene of Autographa catifornica nuclear polyhedrosis virus. Virology 131:561–565
    [Google Scholar]
  16. Jaques R. P. 1975; Persistence, accumulation and denaturation of nuclear polyhedrosis and granulosis viruses. Baculovirusesfor Pest Control: Safety Considerations90–101 Summers M. D., Engler R., Falcon L. A., Vail P. V. Washington, D.C.: American Society for Microbiology;
    [Google Scholar]
  17. Luckow V. A., Summers M. D. 1988; Trends in the development of baculovirus expression vectors. Bio/Technology 6:47–55
    [Google Scholar]
  18. Maeda S., Kawai T., Obinata M., Fujiwara H, Horiuchi t., Saeki Y., Sato Y., Furusawa M. 1985; Production of human alpha interferon in silkworms using a baculovirus vector. Nature, London 315:592–594
    [Google Scholar]
  19. Maniatis T., Fritsch E. F., Sambrook J. 1982 Molecular Cloning: A Laboratory Manual New York: Cold Spring Harbor Laboratory;
    [Google Scholar]
  20. Pennock G. D., Shoemaker C., Miller L. K. 1984; Strong and regulated expression of Escherichia coli beta-galactosidase in insect cells with a baculovirus vector. Molecular and Cellular Biology 4:399–406
    [Google Scholar]
  21. Potter K. N., Miller L. K. 1980; Transfection of two invertebrate cell lines with DNA of Autographa californica NPV. Journal of Invertebrate Pathology 36:431–432
    [Google Scholar]
  22. Smith G. E., Summers M. D., Fraser M. J. 1983; Production of human beta-interferon in insect cells by a baculovirus expression vector. Molecular and Cellular Biology 3:2156–2165
    [Google Scholar]
  23. zelazny B. 1972; Studies on Rhabdionvirus Oryctes. I. Effect on larvae of Oryctes rhinoceros and inactivation of the virus. Journal of Invertebrate Pathology 20:235–241
    [Google Scholar]
  24. Zelazny B., Autar M. L., Singh R., Malone L. A. 1988; Papuana uninodis a new host for the baculovirus of Oryctes. Journal of Invertebrate Pathology 60:157–160
    [Google Scholar]
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