1887

Abstract

Accumulation of baculovirus defective interfering particle (DIP) and few polyhedra (FP) mutants is a major limitation to continuous large-scale baculovirus production in insect-cell culture. Although overcoming these mutations would result in a cheaper platform for producing baculovirus biopesticides, little is known regarding the mechanism of FP and DIP formation. This issue was addressed by comparing DIP production of wild-type (WT) multiple nucleopolyhedrovirus (AcMNPV) with that of a recombinant AcMNPV (denoted Ac-FPm) containing a modified gene with altered transposon insertion sites that prevented transposon-mediated production of the FP phenotype. In addition to a reduction in the incidence of the FP phenotype, DIP formation was delayed on passaging of Ac-FPm compared with WT AcMNPV. Specifically, the yield of DIP DNA in Ac-FPm was significantly lower than in WT AcMNPV up to passage 16, thereby demonstrating that modifying the transposon insertion sites increases the genomic stability of AcMNPV. A critical component of this investigation was the optimization of a systematic method based on the use of pulsed-field gel electrophoresis (PFGE) to characterize extracellular virus DNA. Specifically, PFGE was used to detect defective genomes, determine defective genome sizes and quantify the amount of defective genome within a heterogeneous genome population of passaged virus.

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2012-02-01
2020-11-30
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References

  1. Ayres M. D., Howard S. C., Kuzio J., Lopez-Ferber M., Possee R. D. 1994; The complete DNA sequence of Autographa californica nuclear polyhedrosis virus. Virology 202:586–605 [CrossRef][PubMed]
    [Google Scholar]
  2. Black B. C., Brennan L. A., Dierks P. M., Gard I. E. 1997; Commercialization of baculoviral insecticides. In The Baculoviruses pp. 341–388 Edited by Miller L. K. New York: Plenum Press; [CrossRef]
    [Google Scholar]
  3. Burand J. P., Summers M. D. 1982; Alteration of Autographa californica nuclear polyhedrosis virus DNA upon serial passage in cell culture. Virology 119:223–229 [CrossRef][PubMed]
    [Google Scholar]
  4. Burand J. P., Wood H. A., Summers M. D. 1983; Defective particles from a persistent baculovirus infection in Trichoplusia ni tissue culture cells. J Gen Virol 64:391–398 [CrossRef]
    [Google Scholar]
  5. Carstens E. B. 1987; Identification and nucleotide sequence of the regions of Autographa californica nuclear polyhedrosis virus genome carrying insertion elements derived from Spodoptera frugiperda. . Virology 161:8–17 [CrossRef][PubMed]
    [Google Scholar]
  6. Cochran M. A., Faulkner P. 1983; Location of homologous DNA sequences intercepted at five regions in the baculovirus AcMNPV genome. J Gen Virol 45:961–970
    [Google Scholar]
  7. DuPont 1996; Notification to conduct small-scale field testing of a genetically altered baculovirus. EPA No. 352-NMP-4.
  8. Fraser M. J., Smith G. E., Summers M. D. 1983; Acquisition of host cell DNA sequences by baculoviruses: relationship between host DNA insertions and FP mutants of Autographa californica and Galleria mellonella nuclear polyhedrosis viruses. J Virol 47:287–300[PubMed]
    [Google Scholar]
  9. Gard I. E. 1997; Field testing a genetically modified baculovirus. In Brighton Crop Protection Conference Symposium Proceedings No. 68: Microbial Insecticides: Novelty or Necessity? pp. 101–114
    [Google Scholar]
  10. Giri L., Li H., Sandgren D., Feiss M. G., Roller R., Bonning B. C., Murhammer D. W. 2010; Removal of transposon target sites from the Autographa californica multiple nucleopolyhedrovirus fp25k gene delays, but does not prevent, accumulation of the few polyhedra phenotype. J Gen Virol 91:3053–3064 [CrossRef][PubMed]
    [Google Scholar]
  11. Guarino L. A., Gonzalez M. A., Summers M. D. 1986; Complete sequence and enhancer function of the homologous DNA regions of Autographa californica nuclear polyhedrosis virus. J Virol 60:224–229[PubMed]
    [Google Scholar]
  12. Harrison R. L., Summers M. D. 1995; Mutations in the Autographa californica multinucleocapsid nuclear polyhedrosis virus 25 kDa protein gene result in reduced virion occlusion, altered intranuclear envelopment and enhanced virus production. J Gen Virol 76:1451–1459 [CrossRef][PubMed]
    [Google Scholar]
  13. Jarvis D. L., Garcia A. Jr 1994; Long-term stability of baculoviruses stored under various conditions. Biotechniques 16:508–513[PubMed]
    [Google Scholar]
  14. Kelly B. J., King L. A., Possee R. D. 2007; Introduction to baculovirus molecular biology. In Baculovirus and Insect Cell Expression Protocols pp. 25–53 Edited by Murhammer D. W. Totowa, New Jersey: Humana Press;
    [Google Scholar]
  15. Kompier R., Tramper J., Vlak J. M. 1988; A continuous process for the production of baculovirus using insect-cell cultures. Biotechnol Lett 10:849–854 [CrossRef]
    [Google Scholar]
  16. Kool M., Voncken J. W., van Lier F. L. J., Tramper J., Vlak J. M. 1991; Detection and analysis of Autographa californica nuclear polyhedrosis virus mutants with defective interfering properties. Virology 183:739–746 [CrossRef][PubMed]
    [Google Scholar]
  17. Krell P. J. 1996; Passage effect of virus infection in insect cells. Cytotechnology 20:125–137 [CrossRef]
    [Google Scholar]
  18. Kumar S., Miller L. K. 1987; Effects of serial passage of Autographa californica nuclear polyhedrosis virus in cell culture. Virus Res 7:335–349 [CrossRef][PubMed]
    [Google Scholar]
  19. Lee H. Y., Krell P. J. 1992; Generation and analysis of defective genomes of Autographa californica nuclear polyhedrosis virus. J Virol 66:4339–4347[PubMed]
    [Google Scholar]
  20. Miller D. W., Miller L. K. 1982; A virus mutant with an insertion of a copia-like transposable element. Nature 299:562–564 [CrossRef][PubMed]
    [Google Scholar]
  21. Moscardi F. 1999; Assessment of the application of baculoviruses for control of Lepidoptera. Annu Rev Entomol 44:257–289 [CrossRef][PubMed]
    [Google Scholar]
  22. Murhammer D. W. 1996; Use of viral insecticides for pest control and production in cell culture. Appl Biochem Biotechnol 59:199–220 [CrossRef]
    [Google Scholar]
  23. O’Reilly D. R., Passarelli A. L., Goldman I. F., Miller L. K. 1990; Characterization of the DA26 gene in a hypervariable region of the Autographa californica nuclear polyhedrosis virus genome. J Gen Virol 71:1029–1037 [CrossRef][PubMed]
    [Google Scholar]
  24. Parekh D. 2008 Development and Scale-up of Protein Production Using the Baculovirus Expression System Boston, MA: Process Sciences, Diosynth Biotechnology, Baculovirus Technology;
    [Google Scholar]
  25. Pfaller M. A., Hollis R. J., Sader H. S. 1994; Chromosomal restriction fragment analysis by pulsed field gel electrophoresis. In Clinical Microbiology Procedures HandbookSuppl. 1pp. 10.5c.1–10.5c12 Edited by Isenberg H. D. Washington, DC: American Society for Microbiology;
    [Google Scholar]
  26. Pijlman G. P., van den Born E., Martens D. E., Vlak J. M. 2001; Autographa californica baculoviruses with large genomic deletions are rapidly generated in infected insect cells. Virology 283:132–138 [CrossRef][PubMed]
    [Google Scholar]
  27. Pijlman G. P., Dortmans J. C., Vermeesch A. M. G., Yang K., Martens D. E., Goldbach R. W., Vlak J. M. 2002; Pivotal role of the non-hr origin of DNA replication in the genesis of defective interfering baculoviruses. J Virol 76:5605–5611 [CrossRef][PubMed]
    [Google Scholar]
  28. Pijlman G. P., van Schijndel J. E., Vlak J. M. 2003; Spontaneous excision of BAC vector sequences from bacmid-derived baculovirus expression vectors upon passage in insect cells. J Gen Virol 84:2669–2678 [CrossRef][PubMed]
    [Google Scholar]
  29. Pijlman G. P., de Vrij J., van den End F. J., Vlak J. M., Martens D. E. 2004; Evaluation of baculovirus expression vectors with enhanced stability in continuous cascaded insect-cell bioreactors. Biotechnol Bioeng 87:743–753 [CrossRef][PubMed]
    [Google Scholar]
  30. Potter K. N., Miller L. K. 1980; Correlating genetic mutation of a baculovirus with the physical map of the DNA genome. ICN-UCLA Symp Mol Cell Biol 18:71–80
    [Google Scholar]
  31. Slansky J. 2008 Generation of Antigen-specific Antitumor Immunity Using Insect Cells Infected with Recombinant Baculoviruses Boston, MA: Health Sciences Center, University of Colorado, Baculovirus Technology;
    [Google Scholar]
  32. Tramper J., Vlak J. M. 1986; Some engineering and economic aspects of continuous cultivation of insect cells for the production of baculoviruses. Ann N Y Acad Sci 469:279–288 [CrossRef]
    [Google Scholar]
  33. Vanarsdall A. L., Mikhailov V. S., Rohrmann G. F. 2007; Characterization of a baculovirus lacking the DBP (DNA-binding protein) gene. Virology 364:475–485 [CrossRef][PubMed]
    [Google Scholar]
  34. van Hulten M. C. W., Witteveldt J., Peters S., Kloosterboer N., Tarchini R., Fiers M., Sandbrink H., Lankhorst R. K., Vlak J. M. 2001; The white spot syndrome virus DNA genome sequence. Virology 286:7–22 [CrossRef][PubMed]
    [Google Scholar]
  35. van Lier F. L. J., van den End E. J., de Gooijer C. D., Vlak J. M., Tramper J. 1990; Continuous production of baculovirus in a cascade of insect-cell reactors. Appl Microbiol Biotechnol 33:43–47[PubMed] [CrossRef]
    [Google Scholar]
  36. van Lier F. L. J., van der Meijs W. C., Grobben N. G., Olie R. A., Vlak J. M., Tramper J. 1992; Continuous β-galactosidase production with a recombinant baculovirus insect-cell system in bioreactors. J Biotechnol 22:291–298 [CrossRef][PubMed]
    [Google Scholar]
  37. van Lier F. L. J., van Duijnhoven G. C., de Vaan M. M., Vlak J. M., Tramper J. 1994; Continuous β-galactosidase production in insect cells with a p10 gene based baculovirus vector in a two-stage bioreactor system. Biotechnol Prog 10:60–64 [CrossRef][PubMed]
    [Google Scholar]
  38. Vaughn J. L., Goodwin R. H., Tompkins G. J., McCawley P. 1977; The establishment of two cell lines from the insect Spodoptera frugiperda (Lepidoptera; Noctuidae). In Vitro 13:213–217 [CrossRef][PubMed]
    [Google Scholar]
  39. Vlak J. M., Smith G. E. 1982; Orientation of the genome of Autographa californica nuclear polyhedrosis virus: a proposal. J Virol 41:1118–1121
    [Google Scholar]
  40. Wickham T. J., Davis T., Granados R. R., Hammer D. A., Shuler M. L., Wood H. A. 1991; Baculovirus defective interfering particles are responsible for variations in recombinant protein production as a function of multiplicity of infection. Biotechnol Lett 13:483–488 [CrossRef]
    [Google Scholar]
  41. Zwart M. P., Erro E., van Oers M. M., de Visser J. A. G. M., Vlak J. M. 2008; Low multiplicity of infection in vivo results in purifying selection against baculovirus deletion mutants. J Gen Virol 89:1220–1224 [CrossRef][PubMed]
    [Google Scholar]
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