1887

Abstract

Repeated baculovirus infections in cultured insect cells lead to the generation of defective interfering viruses (DIs), which accumulate at the expense of the intact helper virus and compromise heterologous protein expression. In particular, multicapsid nucleopolyhedovirus (AcMNPV) DIs are enriched in an origin of viral DNA replication () not associated with the homologous regions (s). This non- is located within the coding sequence of the non-essential gene. We investigated the effect of a deletion of the AcMNPV non- on the heterologous protein expression levels following serial passage in Sf21 insect cells. Using homologous ET recombination in , deletions within the gene were made in a bacterial artificial chromosome (BAC) containing the entire AcMNPV genome (bacmid). All bacmids were equipped with an expression cassette containing the green fluorescent protein gene and a gene encoding the classical swine fever virus E2 glycoprotein (CSFV-E2). For the parental (intact) bacmid only, a strong accumulation of DIs with reiterated non-s was observed. This was not observed for the mutants, indicating that removal of the non- enhanced the genetic stability of the viral genome upon passaging. However, for all passaged viruses it was found that the entire BAC vector including the expression cassette was spontaneously deleted from the viral genome, leading to a rapid decrease in GFP and CSFV-E2 production. The rationale for the (intrinsic) genetic instability of the BAC vector in insect cells and the implications with respect to large-scale production of proteins with bacmid-derived baculoviruses are discussed.

Loading

Article metrics loading...

/content/journal/jgv/10.1099/vir.0.19438-0
2003-10-01
2020-07-07
Loading full text...

Full text loading...

/deliver/fulltext/jgv/84/10/vir842669.html?itemId=/content/journal/jgv/10.1099/vir.0.19438-0&mimeType=html&fmt=ahah

References

  1. Adler H., Messerle M., Koszinowski U. H.. 2001; Virus reconstituted from infectious bacterial artificial chromosome (BAC)-cloned murine gammaherpesvirus 68 acquires wild-type properties in vivo only after excision of BAC vector sequences. J Virol75:5692–5696
    [Google Scholar]
  2. Altschul S. F., Madden T. L., Schaffer A. A., Zhang J., Zhang Z., Miller W., Lipman D. J.. 1997; Gapped BLAST and PSI-BLAST: a new generation of protein database search programs. Nucleic Acids Res25:3389–3402
    [Google Scholar]
  3. 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. Virology202:586–605
    [Google Scholar]
  4. Bett A. J., Prevec L., Graham F. L.. 1993; Packaging capacity and stability of human adenovirus type 5 vectors. J Virol67:5911–5921
    [Google Scholar]
  5. Bigot Y., Stasiak K., Rouleux-Bonnin F., Federici B. A.. 2000; Characterization of repetitive DNA regions and methylated DNA in ascovirus genomes. J Gen Virol81:3073–3082
    [Google Scholar]
  6. Bloss T. A., Sugden B.. 1994; Optimal lengths for DNAs encapsidated by Epstein–Barr virus. J Virol68:8217–8222
    [Google Scholar]
  7. Brune W., Messerle M., Koszinowski U. H.. 2000; Forward with BACs – new tools for herpesvirus genomics. Trends Genet16:254–259
    [Google Scholar]
  8. Clem R. J., Robson M., Miller L. K.. 1994; Influence of infection route on the infectivity of baculovirus mutants lacking the apoptosis-inhibiting gene p35 and the adjacent gene p94. J Virol68:6759–6762
    [Google Scholar]
  9. Dai X. J., 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 Virol81:2545–2554
    [Google Scholar]
  10. Davis S. J., Vierstra R. D.. 1998; Soluble, highly fluorescent variants of green fluorescent protein (GFP) for use in higher plants. Plant Mol Biol36:521–528
    [Google Scholar]
  11. Friesen P. D.. 1997; Regulation of baculovirus early gene expression. In The Baculoviruses pp 141–170 Edited by Miller L. K.. New York: Plenum Press;
    [Google Scholar]
  12. Friesen P. D., Miller L. K.. 1987; Divergent transcription of early 35- and 94-kilodalton protein genes encoded by the Hin dIII K genome fragment of the baculovirus Autographa californica nuclear polyhedrosis virus. J Virol61:2264–2272
    [Google Scholar]
  13. Friesen P. D., Nissen M. S.. 1990; Gene organization and transcription of TED, a lepidopteran retrotransposon integrated within the baculovirus genome. Mol Cell Biol10:3067–3077
    [Google Scholar]
  14. Hayakawa T., Ko R., Okano K., Seong S. I., Goto C., Maeda S.. 1999; Sequence analysis of the Xestia c-nigrum granulovirus genome. Virology262:277–297
    [Google Scholar]
  15. Heldens J. G., 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 Virol77:3127–3134
    [Google Scholar]
  16. Heldens J. G., Broer R., Zuidema D., Goldbach R. W., Vlak J. M.. 1997; Identification and functional analysis of a non- hr origin of DNA replication in the genome of Spodoptera exigua multicapsid nucleopolyhedrovirus. J Gen Virol78:1497–1506
    [Google Scholar]
  17. Hu Z. H., Arif B. M., Sun J. S., Chen X. W., Zuidema D., Goldbach R. W., Vlak J. M.. 1998; Genetic organization of the Hin dIII-I region of the single-nucleocapsid nucleopolyhedrovirus of Buzura suppressaria . Virus Res55:71–82
    [Google Scholar]
  18. Huang J., Levin D. B.. 1999; Identification and functional analysis of a putative non- hr origin of DNA replication from the Spodoptera littoralis type B multinucleocapsid nucleopolyhedrovirus. J Gen Virol80:2263–2274
    [Google Scholar]
  19. IJkel W. F., van Strien E. A., Heldens J. G., Broer R., Zuidema D., Goldbach R. W., Vlak J. M.. 1999; Sequence and organization of the Spodoptera exigua multicapsid nucleopolyhedrovirus genome. J Gen Virol80:3289–3304
    [Google Scholar]
  20. Jehle J. A.. 2002; The expansion of a hypervariable, non- hr ori -like region in the genome of Cryptophlebia leucotreta granulovirus provides in vivo evidence for the utilization of baculovirus non- hr ori s during replication. J Gen Virol83:2025–2034
    [Google Scholar]
  21. Kamita S. G., Majima K., Maeda S.. 1993; Identification and characterization of the p35 gene of Bombyx mori nuclear polyhedrosis virus that prevents virus-induced apoptosis. J Virol67:455–463
    [Google Scholar]
  22. King L. A., Possee R. D.. 1992; The baculovirus expression system Dordrecht: Kluwer;
    [Google Scholar]
  23. Kitts P. A.. 1996; Construction of baculovirus recombinants. Cytotechnology20:111–123
    [Google Scholar]
  24. Kool M., Voncken J. W., van Lier F. L., Tramper J., Vlak J. M.. 1991; Detection and analysis of Autographa californica nuclear polyhedrosis virus mutants with defective interfering properties. Virology183:739–746
    [Google Scholar]
  25. Kool M., van den Berg P. M., Tramper J., Goldbach R. W., Vlak J. M.. 1993; Location of two putative origins of DNA replication of Autographa californica nuclear polyhedrosis virus. Virology192:94–101
    [Google Scholar]
  26. Kool M., Goldbach R. W., Vlak J. M.. 1994; A putative non- hr origin of DNA replication in the Hin dIII-K fragment of Autographa californica multiple nucleocapsid nuclear polyhedrosis virus. J Gen Virol75:3345–3352
    [Google Scholar]
  27. Krell P. J.. 1996; Passage effect of virus infection in insect cells. Cytotechnology20:125–137
    [Google Scholar]
  28. Kumar S., Miller L. K.. 1987; Effects of serial passage of Autographa californica nuclear polyhedrosis virus in cell culture. Virus Res7:335–349
    [Google Scholar]
  29. Lee H. Y., Krell P. J.. 1992; Generation and analysis of defective genomes of Autographa californica nuclear polyhedrosis virus. J Virol66:4339–4347
    [Google Scholar]
  30. Lee H., Krell P. J.. 1994; Reiterated DNA fragments in defective genomes of Autographa californica nuclear polyhedrosis virus are competent for AcMNPV-dependent DNA replication. Virology202:418–429
    [Google Scholar]
  31. Lu A., Krell P. J., Vlak J. M., Rohrmann G. F.. 1997; Baculovirus DNA replication. In The Baculoviruses pp 171–216 Edited by Miller L. K.. New York: Plenum Press;
    [Google Scholar]
  32. 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 Virol67:4566–4579
    [Google Scholar]
  33. Luque T., Finch R., Crook N., O'Reilly D. R., Winstanley D.. 2001; The complete sequence of the Cydia pomonella granulovirus genome. J Gen Virol82:2531–2547
    [Google Scholar]
  34. Muyrers J. P., Zhang Y., Testa G., Stewart A. F.. 1999; Rapid modification of bacterial artificial chromosomes by ET-recombination. Nucleic Acids Res27:1555–1557
    [Google Scholar]
  35. Pearson M. N., Bjornson R. M., Ahrens C., Rohrmann G. F.. 1993; Identification and characterization of a putative origin of DNA replication in the genome of a baculovirus pathogenic for Orgyia pseudotsugata . Virology197:715–725
    [Google Scholar]
  36. 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. Virology283:132–138
    [Google Scholar]
  37. Pijlman G. P., Dortmans J. C. F. M., 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 Virol76:5605–5611
    [Google Scholar]
  38. Possee R. D.. 1997; Baculoviruses as expression vectors. Curr Opin Biotechnol8:569–572
    [Google Scholar]
  39. Roosien J., Usmany M., Klinge-Roode E. C., Meijerink P. H. S., Vlak J. M.. 1986; Heterologous recombination between baculoviruses. In Fundamental and Applied Aspects of Invertebrate Pathology pp 389–392 Edited by Samson R. A., Vlak J. M., Peters D.. ICIP86 Foundation
    [Google Scholar]
  40. Sambrook J., Fritsch E. F., Maniatis T.. 1989; Molecular Cloning: a Laboratory Manual , 2nd edn. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory;
    [Google Scholar]
  41. Shizuya H., Birren B., Kim U. J., Mancino V., Slepak T., Tachiiri Y., Simon M.. 1992; Cloning and stable maintenance of 300-kilobase-pair fragments of human DNA in Escherichia coli using an F-factor-based vector. Proc Natl Acad Sci U S A89:8794–8797
    [Google Scholar]
  42. Smith G. A., Enquist L. W.. 1999; Construction and transposon mutagenesis in Escherichia coli of a full-length infectious clone of pseudorabies virus, an alphaherpesvirus. J Virol73:6405–6414
    [Google Scholar]
  43. Smith G. A., Enquist L. W.. 2000; A self-recombining bacterial artificial chromosome and its application for analysis of herpesvirus pathogenesis. Proc Natl Acad Sci U S A97:4873–4878
    [Google Scholar]
  44. Summers M. D., Smith G. E.. 1987; A Manual of Methods for Baculovirus Vectors and Insect Cell Culture Procedures . Texas Agricultural Experiment Station Bulletin No. 1555
    [Google Scholar]
  45. 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. Virology286:7–22
    [Google Scholar]
  46. Van Oers M. M., Thomas A. A. M., Moormann R. J. M., Vlak J. M.. 2001; Secretory pathway limits the enhanced expression of classical swine fever virus E2 glycoprotein in insect cells. J Biotechnol86:31–38
    [Google Scholar]
  47. Van Rijn P. A., van Gennip H. G., Moormann R. J.. 1999; An experimental marker vaccine and accompanying serological diagnostic test both based on envelope glycoprotein E2 of classical swine fever virus (CSFV). Vaccine17:433–440
    [Google Scholar]
  48. 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 Vitro13:213–217
    [Google Scholar]
  49. Vialard J. E., Arif B. M., Richardson C. D.. 1995; Introduction to the molecular biology of baculoviruses. Methods Mol Biol39:1–24
    [Google Scholar]
  50. Vlak J. M.. 1979; The proteins of nonoccluded Autographa californica nuclear polyhedrosis virus produced in an established cell line of Spodoptera frugiperda . J Invertebr34:110–118
    [Google Scholar]
  51. Wagner M., Jonjic S., Koszinowski U. H., Messerle M.. 1999; Systematic excision of vector sequences from the BAC-cloned herpesvirus genome during virus reconstitution. J Virol73:7056–7060
    [Google Scholar]
  52. 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 Lett13:483–488
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jgv/10.1099/vir.0.19438-0
Loading
/content/journal/jgv/10.1099/vir.0.19438-0
Loading

Data & Media loading...

Most cited this month Most Cited RSS feed

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