1887

Abstract

Rapid accumulation of few polyhedra (FP) mutants was detected during serial passaging of nucleopolyhedrovirus (HaSNPV) in cell culture. 100% FP infected cells were observed by passage 6. The specific yield decreased from 178 polyhedra per cell at passage 2 to two polyhedra per cell at passage 6. The polyhedra at passage 6 were not biologically active, with a 28-fold reduction in potency compared to passage 3. Electron microscopy studies revealed that very few polyhedra were produced in an FP infected cell (<10 polyhedra per section) and in most cases these polyhedra contained no virions. A specific failure in the intranuclear nucleocapsid envelopment process in the FP infected cells, leading to the accumulation of naked nucleocapsids, was observed. Genomic restriction endonuclease digestion profiles of budded virus DNA from all passages did not indicate any large DNA insertions or deletions that are often associated with such FP phenotypes for the extensively studied nucleopolyhedrovirus and nucleopolyhedrovirus. Within an HaSNPV gene homologue, a single base-pair insertion (an adenine residue) within a region of repetitive sequences (seven adenine residues) was identified in one plaque-purified HaSNPV FP mutant. Furthermore, the sequences obtained from individual clones of the gene PCR products of a late passage revealed point mutations or single base-pair insertions occurring throughout the gene. The mechanism of FP mutation in HaSNPV is likely similar to that seen for nucleopolyhedrovirus, involving point mutations or small insertions/deletions of the gene.

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2002-04-01
2020-01-27
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References

  1. Ahrens, C. H., Russell, R. L., Funk, C. J., Evans, J. T., Harwood, S. H. & Rohrmann, G. F. ( 1997; ). The sequence of the Orgyia pseudotsugata multinucleocapsid nuclear polyhedrosis virus genome. Virology 229, 381-399.[CrossRef]
    [Google Scholar]
  2. 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.[CrossRef]
    [Google Scholar]
  3. Beames, B. & Summers, M. D. ( 1988; ). Comparisons of host cell DNA insertions and altered transcription at the site of insertions in few polyhedra baculovirus mutants. Virology 162, 206-220.[CrossRef]
    [Google Scholar]
  4. Beames, B. & Summers, M. D. ( 1989; ). Location and nucleotide sequence of the 25K protein missing from baculovirus few polyhedra (FP) mutants. Virology 168, 344-353.[CrossRef]
    [Google Scholar]
  5. Beniya, H., Braunagel, S. C. & Summers, M. D. ( 1998; ). Autographa californica nuclear polyhedrosis virus: subcellular localization and protein trafficking of BV/ODV-E26 to intranuclear membranes and viral envelopes. Virology 240, 64-75.[CrossRef]
    [Google Scholar]
  6. Bischoff, D. S. & Slavicek, J. M. ( 1997; ). Phenotypic and genotypic analysis of Lymantria dispar nucleopolyhedrovirus few polyhedra mutants: mutations in 25K FP gene may be caused by DNA replication errors. Journal of Virology 71, 1097-1106.
    [Google Scholar]
  7. Braunagel, S. C., Burks, J. K., Rosas-Acosta, G., Harrison, R. L., Ma, H. & Summers, M. D. ( 1999; ). Mutations within the Autographa californica nucleopolyhedrovirus FP25K gene decrease the accumulation of ODV-E66 and alter its intranuclear transport. Journal of Virology 73, 8559-8570.
    [Google Scholar]
  8. Chakraborty, S. (1997). In vitro production of wild-type Helicoverpa baculovirus. PhD thesis, University of Queensland, Australia.
  9. Chakraborty, S. & Reid, S. ( 1999; ). Serial passage of a Helicoverpa armigera nucleopolyhedrovirus in Helicoverpa zea cell cultures. Journal of Invertebrate Pathology 73, 303-308.[CrossRef]
    [Google Scholar]
  10. Chen, X., IJkel, F. W. J., Tarchini, R., Sun, X., Sandbrink, H., Wang, H., Peters, S., Zuidema, D., Lankhorst, R. K., Vlak, J. M. & Hu, Z. ( 2001; ). The sequence of the Helicoverpa armigera single nucleocapsid nucleopolyhedrovirus genome. Journal of General Virology 82, 241-257.
    [Google Scholar]
  11. Fraser, M. J. & Hink, W. F. ( 1982; ). Comparative sensitivity of several plaque assay techniques employing TN-368 and IPLB-SF 21AE insect cell lines for plaque variants of Galleria mellonella nuclear polyhedrosis virus. Journal of Invertebrate Pathology 40, 89-97.[CrossRef]
    [Google Scholar]
  12. 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. Journal of Virology 67, 287-300.
    [Google Scholar]
  13. Fraser, M. J., Brusca, J. S., Smith, G. E. & Summers, M. D. ( 1985; ). Transposon-mediated mutagenesis of a baculovirus. Virology 145, 356-361.[CrossRef]
    [Google Scholar]
  14. Gomi, S., Majima, K. & Maeda, S. ( 1999; ). Sequence analysis of the genome of Bombyx mori nucleopolyhedrovirus. Journal of General Virology 80, 1323-1337.
    [Google Scholar]
  15. Harrap, K. A. ( 1972; ). The structure of nuclear polyhedrosis viruses. III. Virus assembly. Virology 50, 133-139.[CrossRef]
    [Google Scholar]
  16. Harrison, R. L. & Summers, M. D. ( 1995a; ). Biosynthesis and localization of the Autographa californica nuclear polyhedrosis virus 25K gene product. Virology 208, 279-288.[CrossRef]
    [Google Scholar]
  17. Harrison, R. L. & Summers, M. D. ( 1995b; ). 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. Journal of General Virology 76, 1451-1459.[CrossRef]
    [Google Scholar]
  18. Harrison, R. L., Jarvis, D. L. & Summers, M. D. ( 1996; ). The role of the AcMNPV 25K gene, ‘FP25,’ in baculovirus polh and p10 expression. Virology 226, 34-46.[CrossRef]
    [Google Scholar]
  19. Hink, W. F. & Vail, P. V. ( 1973; ). A plaque assay for titration of alfalfa looper nuclear polyhedrosis virus in a cabbage looper (TN-368) cell line. Journal of Invertebrate Pathology 22, 168-174.[CrossRef]
    [Google Scholar]
  20. IJkel, W. F. J., Van Strien, E. A., Heldens, J. G. M., Broer, R., Zuidema, D., Goldbach, R. W. & Vlak, J. M. ( 1999; ). Sequence and organization of the Spodoptera exigua multicapsid nucleopolyhedrovirus genome. Journal of General Virology 80, 3289-3304.
    [Google Scholar]
  21. Katsuma, S., Noguchi, Y., Zhou, C. L., Kobayashi, M. & Maeda, S. ( 1999; ). Characterization of the 25K FP gene of the baculovirus Bombyx mori nucleopolyhedrovirus: implications for post-mortem host degradation. Journal General Virology 80, 783-91.
    [Google Scholar]
  22. Lua, L. H. L. & Reid, S. R. ( 2000; ). Virus morphogenesis of Helicoverpa armigera nucleopolyhedrovirus in Helicoverpa zea serum-free suspension culture. Journal of General Virology 81, 2531-2543.
    [Google Scholar]
  23. Lynn, D. E. ( 1994; ). Enhanced infectivity of occluded virions of the gypsy moth nuclear polyhedrosis virus for cell culture. Journal of Invertebrate Pathology 63, 268-274.[CrossRef]
    [Google Scholar]
  24. McIntosh, A. H. & Ignoffo, C. M. ( 1981; ). Replication and infectivity of the single-embedded nuclear polyhedrosis virus, baculovirus Heliothis, in homologous cell line. Journal of Invertebrate Pathology 37, 258-264.[CrossRef]
    [Google Scholar]
  25. Miller, L. K. ( 1986; ). The genetics of baculoviruses. In The Biology of Baculoviruses , pp. 217-238. Edited by R. R. Granados & B. A. Federici. Florida:CRC Press.
  26. Nielsen, L. K., Smyth, G. K. & Greenfield, P. F. ( 1991; ). Hemocytometer cell count distributions: implications of non-Poisson behavior. Biotechnology Progress 7, 560-563.[CrossRef]
    [Google Scholar]
  27. Potter, K. N., Faulkner, P. & MacKinnon, E. A. ( 1976; ). Strain selection during serial passage of Trichoplusia ni nuclear polyhedrosis virus. Journal of Virology 18, 1040-1050.
    [Google Scholar]
  28. Potter, K. N., Jaques, R. P. & Faulkner, P. ( 1978; ). Modification of Trichoplusia ni nuclear polyhedrosis virus passaged in vivo. Intervirology 9, 76-85.[CrossRef]
    [Google Scholar]
  29. Ramoska, W. A. & Hink, W. F. ( 1974; ). Electron microscope examination of two plaque variants from a nuclear polyhedrosis virus of the alfalfa looper, Autographa californica. Journal of Invertebrate Pathology 23, 197-201.[CrossRef]
    [Google Scholar]
  30. Rhodes, D. J. ( 1996; ). Economics of baculovirus–insect cell production systems. Cytotechnology 20, 291-297.[CrossRef]
    [Google Scholar]
  31. Russell, R. L. Q. & Rohrmann, G. F. ( 1993; ). A 25 kDa protein is associated with the envelopes of occluded baculovirus virions. Journal of General Virology 195, 532-540.
    [Google Scholar]
  32. Slavicek, J. M., Podgwaite, J. & Lanner-Herrera, C. ( 1992; ). Properties of two Lymantria dispar nuclear polyhedrosis virus isolates obtained from microbial pesticide Gypchek. Journal of Invertebrate Pathology 59, 142-148.[CrossRef]
    [Google Scholar]
  33. Slavicek, J. M., Hayes-Plazolles, N. & Kelly, M. E. ( 1995; ). Rapid formation of few polyhedra mutants of Lymantria dispar multinucleocapsid nuclear polyhedrosis virus during serial passage in cell culture. Biological Control 5, 251-261.[CrossRef]
    [Google Scholar]
  34. Sohi, S. S., Percy, J., Arif, B. M. & Cunningham, J. C. ( 1984; ). Replication and serial passage of a singly enveloped baculovirus of Orgyia leucostigma in homologous cell lines. Intervirology 21, 50-60.[CrossRef]
    [Google Scholar]
  35. Taylor, J. R. (1997). The binomial distribution. In An Introduction to Error Analysis: The Study of Uncertainties in Physical Measurements, 2nd edn, pp. 227–244. California: University Science Books.
  36. Wang, H. H., Fraser, M. J. & Cary, L. C. ( 1989; ). Transposon mutagenesis of baculoviruses: analysis of TFP3 lepidopteran transposon insertions at the FP locus of nuclear polyhedrosis virus. Gene 81, 97-108.[CrossRef]
    [Google Scholar]
  37. Wood, H. A. ( 1980; ). Isolation and replication of an occlusion body-deficient mutant of the Autographa californica nuclear polyhedrosis virus. Virology 105, 338-344.[CrossRef]
    [Google Scholar]
  38. Yamada, K., Sherman, K. E. & Maramorosch, K. ( 1982; ). Serial passage of Heliothis zea singly embedded nuclear polyhedrosis virus in a homologous cell line. Journal of Invertebrate Pathology 39, 185-191.[CrossRef]
    [Google Scholar]
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