1887

Abstract

The fate of multiple nucleopolyhedrovirus deletion mutants originating from serial passage in cell culture was investigated by passaging a population enriched in these mutants in insect larvae. The infectivity of polyhedra and occlusion-derived virion content per polyhedron were restored within two passages . The frequency of occurrence of deletion mutants was determined by real-time PCR. The frequency of the non-homologous region origin (non-HR ) of DNA replication was reduced to wild-type levels within two passages. The frequency of the polyhedrin gene did not increase and remained below wild-type levels. A low m.o.i. during the initial infection in insect larvae, causing strong purifying selection for autonomously replicating viruses, could explain these observations. The same virus population used was also passaged once at a different m.o.i. in cell culture. A similar effect (i.e. lower non-HR frequency) was observed at low m.o.i. only, indicating that m.o.i. was the key selective condition.

Loading

Article metrics loading...

/content/journal/jgv/10.1099/vir.0.83645-0
2008-05-01
2019-11-18
Loading full text...

Full text loading...

/deliver/fulltext/jgv/89/5/1220.html?itemId=/content/journal/jgv/10.1099/vir.0.83645-0&mimeType=html&fmt=ahah

References

  1. Bull, J. C., Godfray, H. C. J. & O'Reilly, D. R. ( 2001; ). Persistence of an occlusion-negative recombinant nucleopolyhedrovirus in Trichoplusia ni indicates high multiplicity of cellular infection. Appl Environ Microbiol 67, 5204–5209.[CrossRef]
    [Google Scholar]
  2. Bull, J. C., Godfray, H. C. J. & O'Reilly, D. R. ( 2003; ). A few-polyhedra mutant and wild-type nucleopolyhedrovirus remain as a stable polymorphism during serial coinfection in Trichoplusia ni. Appl Environ Microbiol 69, 2052–2057.[CrossRef]
    [Google Scholar]
  3. Dai, X., Hajós, J. P., Joosten, N. N., van Oers, M. M., IJkel, W. F. J., Zuidema, D., Pang, Y. & Vlak, J. M. ( 2000; ). Isolation of a Spodoptera exigua baculovirus recombinant with a 10.6 kbp genome deletion that retains biological activity. J Gen Virol 81, 2545–2554.
    [Google Scholar]
  4. de Gooijer, C. D., Koken, R. H. M., van Lier, F. L. J., Kool, M., Vlak, J. M. & Tramper, J. ( 1992; ). A structured dynamic model for the baculovirus infection process in insect-cell reactor configurations. Biotechnol Bioeng 40, 537–548.[CrossRef]
    [Google Scholar]
  5. Heldens, J. G. M., van Strien, E. A., Feldmann, A. M., Kulcsar, P., Muñoz, D., Leisy, D. J., Zuidema, D., Goldbach, R. W. & Vlak, J. M. ( 1996; ). Spodoptera exigua multicapsid nucleopolyhedrovirus deletion mutants generated in cell culture lack virulence in vivo. J Gen Virol 77, 3127–3134.[CrossRef]
    [Google Scholar]
  6. Hughes, P. R. & Wood, H. A. ( 1981; ). A synchronous peroral technique for the bioassay of insect viruses. J Invertebr Pathol 37, 154–159.[CrossRef]
    [Google Scholar]
  7. 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]
    [Google Scholar]
  8. Kovacs, G. R., Choi, J., Guarino, L. A. & Summers, M. D. ( 1992; ). Functional dissection of the Autographa californica nuclear polyhedrosis virus immediate early-1 transcriptional regulatory protein. J Virol 66, 7429–7437.
    [Google Scholar]
  9. Li, W. H. ( 1997; ). Molecular Evolution. Sunderland, MA: Sinauer Associates.
  10. López-Ferber, M., Simón, O., Williams, T. & Caballero, P. ( 2003; ). Defective or effective? Mutualistic interactions between virus genotypes. Proc R Soc Lond B Biol Sci 270, 2249–2255.[CrossRef]
    [Google Scholar]
  11. Luckow, V. A., Lee, S. C., Barry, G. F. & Olins, P. O. ( 1993; ). Efficient generation of infectious recombinant baculoviruses by site-specific transposon-mediated insertion of foreign genes into a baculovirus genome propagated in Escherichia coli. J Virol 67, 4566–4579.
    [Google Scholar]
  12. 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]
    [Google Scholar]
  13. Pijlman, G. P., Dortmans, J., 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]
    [Google Scholar]
  14. 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]
    [Google Scholar]
  15. Smith, I. R. L. & Crook, N. E. ( 1988; ). In vivo isolation of baculovirus genotypes. Virology 166, 240–244.[CrossRef]
    [Google Scholar]
  16. Smith, G. E., Vlak, J. M. & Summers, M. D. ( 1983; ). Physical analysis of Autographa californica nuclear polyhedrosis virus transcripts for polyhedrin and 10,000-molecular-weight protein. J Virol 45, 215–225.
    [Google Scholar]
  17. Smits, P. H., Vandevrie, M. & Vlak, J. M. ( 1986; ). Oviposition of beet armyworm (Lepidoptera, Noctuidae) on greenhouse crops. Environ Entomol 15, 1189–1191.[CrossRef]
    [Google Scholar]
  18. van Lier, F. L. J., van der Meijs, W. C. J., 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]
    [Google Scholar]
  19. van Lier, F. L. J., van den Hombergh, J. P. T. W., de Gooijer, C. D., den Boer, M. M., Vlak, J. M. & Tramper, J. ( 1996; ). Long-term semi-continuous production of recombinant baculovirus protein in a repeated (fed-)batch two-stage reactor system. Enzyme Microb Technol 18, 460–466.[CrossRef]
    [Google Scholar]
  20. Vaughn, J. L., Goodwin, R. H., Tompkins, G. J. & McCawley, P. ( 1977; ). Establishment of two cell lines from insect Spodoptera frugiperda (Lepidoptera-Noctuidae). In Vitro 13, 213–217.[CrossRef]
    [Google Scholar]
  21. von Magnus, P. ( 1954; ). Incomplete forms of influenza virus. Adv Virus Res 2, 59–79.
    [Google Scholar]
  22. 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]
  23. Zwart, M. P., van Oers, M. M., Cory, J. S., van Lent, J. W. M., van der Werf, W. & Vlak, J. M. ( 2008; ). Development of a quantitative real-time PCR for determination of genotype frequencies for studies in baculovirus population biology. J Virol Methods 148, 146–154.[CrossRef]
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jgv/10.1099/vir.0.83645-0
Loading
/content/journal/jgv/10.1099/vir.0.83645-0
Loading

Data & Media loading...

Most Cited This Month

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error