1887

Abstract

In this study, we have deleted the gene of the baculovirus BmNPV, which encodes one of the viral RNA polymerase subunits, in order to create a knockout bacmid, Δ directing cytopathology-free single-cell infections for gene transduction and recombinant protein production. However, while removal of the complete ORF produced the expected phenotype, it also affected the function of the closely linked essential gene thus hampering the mutant bacmid rescue in cultured cells expressing recombinant LEF8. Subsequently, we determined that several diverse sequences can substitute for the 5′-upstream sequences that were removed by the deletion of the gene and also showed that neither a physical linkage nor expression of the two relevant genes under native promoter control is a prerequisite for a fully functional virus. Based on these findings, we generated a rescue-competent -null vector, which contained a heterologous promoter-driven This deficient vector, which produces productive infections and progeny virus lacking in deficiency-complementing cells expressing LEF8, could be used as the basis for an alternative to current silkmoth transduction systems.

Loading

Article metrics loading...

/content/journal/jgv/10.1099/jgv.0.000383
2016-03-01
2024-04-26
Loading full text...

Full text loading...

/deliver/fulltext/jgv/97/3/786.html?itemId=/content/journal/jgv/10.1099/jgv.0.000383&mimeType=html&fmt=ahah

References

  1. Altschul S. F., Gish W., Miller W., Myers E. W., Lipman D. J. 1990; Basic local alignment search tool. J Mol Biol 215:403–410 [View Article][PubMed]
    [Google Scholar]
  2. Bailey T. L., Boden M., Buske F. A., Frith M., Grant C. E., Clementi L., Ren J., Li W. W., Noble W. S. 2009; MEME SUITE: tools for motif discovery and searching. Nucleic Acids Res 37:W202–W208 [View Article][PubMed]
    [Google Scholar]
  3. Benedict M. Q., Scott J. A., Cockburn A. F. 1994; High-level expression of the bacterial opd gene in Drosophila melanogaster: improved inducible insecticide resistance. Insect Mol Biol 3:247–252 [View Article][PubMed]
    [Google Scholar]
  4. Bézier A., Annaheim M., Herbinière J., Wetterwald C., Gyapay G., Bernard-Samain S., Wincker P., Roditi I., Heller M., other authors. 2009; Polydnaviruses of braconid wasps derive from an ancestral nudivirus. Science 323:926–930 [View Article][PubMed]
    [Google Scholar]
  5. Chen Y. R., Zhong S., Fei Z., Hashimoto Y., Xiang J. Z., Zhang S., Blissard G. W. 2013; The transcriptome of the baculovirus Autographa californica multiple nucleopolyhedrovirus in Trichoplusia ni cells. J Virol 87:6391–6405 [CrossRef]
    [Google Scholar]
  6. Chung C. T., Niemela S. L., Miller R. H. 1989; One-step preparation of competent Escherichia coli: transformation and storage of bacterial cells in the same solution. Proc Natl Acad Sci U S A 86:2172–2175 [View Article][PubMed]
    [Google Scholar]
  7. Chen Y. R., Zhong S., Fei Z., Hashimoto Y., Xiang J. Z., Zhang S., Blissard G. W. 2013; The transcriptome of the baculovirus Autographa californica multiple nucleopolyhedrovirus in Trichoplusia ni cells. J Virol 87:6391–6405 [View Article][PubMed]
    [Google Scholar]
  8. Cléry A., Blatter M., Allain F. H. 2008; RNA recognition motifs: boring? Not quite. Curr Opin Struct Biol 18:290–298 [View Article][PubMed]
    [Google Scholar]
  9. Datsenko K. A., Wanner B. L. 2000; One-step inactivation of chromosomal genes in Escherichia coli K-12 using PCR products. Proc Natl Acad Sci U S A 97:6640–6645 [View Article][PubMed]
    [Google Scholar]
  10. Douris V., Swevers L., Labropoulou V., Andronopoulou E., Georgoussi Z., Iatrou K. 2006; Stably transformed insect cell lines: tools for expression of secreted and membrane-anchored proteins and high-throughput screening platforms for drug and insecticide discovery. Adv Virus Res 68:113–156 [View Article][PubMed]
    [Google Scholar]
  11. Efrose R., Swevers L., Iatrou K. 2010; Baculoviruses deficient in ie1 gene function abrogate viral gene expression in transduced mammalian cells. Virology 406:293–301 [View Article][PubMed]
    [Google Scholar]
  12. Farrell P. J. 1998 The development of stably transformed lepidopteran insect cell technology for both the expression of recombinant proteins and the generation of baculovirus artificial chromosomes PhD thesis University of Calgary; Calgary, AB, Canada:
    [Google Scholar]
  13. Georgomanolis T., Iatrou K., Swevers L. 2009; BmCAP, a silkmoth gene encoding multiple protein isoforms characterized by SoHo and SH3 domains: expression analysis during ovarian follicular development. Insect Biochem Mol Biol 39:892–902 [View Article][PubMed]
    [Google Scholar]
  14. Gomi S., Zhou C. E., Yih W., Majima K., Maeda S. 1997; Deletion analysis of four of eighteen late gene expression factor gene homologues of the baculovirus, BmNPV. Virology 230:35–47 [View Article][PubMed]
    [Google Scholar]
  15. Gomi S., Majima K., Maeda S. 1999; Sequence analysis of the genome of Bombyx mori nucleopolyhedrovirus. J Gen Virol 80:1323–1337 [View Article][PubMed]
    [Google Scholar]
  16. Grace T. D. 1967; Establishment of a line of cells from the silkworm Bombyx mori . Nature 216:613 [View Article][PubMed]
    [Google Scholar]
  17. Guarino L. A., Xu B., Jin J., Dong W. 1998; A virus-encoded RNA polymerase purified from baculovirus-infected cells. J Virol 72:7985–7991[PubMed]
    [Google Scholar]
  18. Hall T. A. 1999; BioEdit: a user-friendly biological sequence alignment editor and analysis program for Windows 95/98/NT. In Nucl Acids Symp Ser 41:95–98
    [Google Scholar]
  19. Iatrou K., Swevers L. 2005; Transformed lepidopteran cells expressing a protein of the silkmoth fat body display enhanced susceptibility to baculovirus infection and produce high titers of budded virus in serum-free media. J Biotechnol 120:237–250 [View Article][PubMed]
    [Google Scholar]
  20. Invitrogen 2004 Bac-to-Bac® Baculovirus Expression System (version D, 10359)
  21. Johnson R., Meidinger R. G., Iatrou K. 1992; A cellular promoter-based expression cassette for generating recombinant baculoviruses directing rapid expression of passenger genes in infected insects. Virology 190:815–823 [View Article][PubMed]
    [Google Scholar]
  22. Lu M., Iatrou K. 1996; The genes encoding the P39 and CG30 proteins of Bombyx mori nuclear polyhedrosis virus. J Gen Virol 77:3135–3143 [View Article][PubMed]
    [Google Scholar]
  23. Lu M., Swevers L., Iatrou K. 1998; The p95 gene of Bombyx mori nuclear polyhedrosis virus: temporal expression and functional properties. J Virol 72:4789–4797[PubMed]
    [Google Scholar]
  24. 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[PubMed]
    [Google Scholar]
  25. Marchler-Bauer A., Lu S., Anderson J. B., Chitsaz F., Derbyshire M. K., DeWeese-Scott C., Fong J. H., Geer L. Y., Geer R. C., other authors. 2011; CDD: a Conserved Domain Database for the functional annotation of proteins. Nucleic Acids Res 39:D225–D229 [View Article][PubMed]
    [Google Scholar]
  26. Marshall O. J. 2004; PerlPrimer: cross-platform, graphical primer design for standard, bisulphite and real-time PCR. Bioinformatics 20:2471–2472 [View Article][PubMed]
    [Google Scholar]
  27. Motohashi T., Shimojima T., Fukagawa T., Maenaka K., Park E. Y. 2005; Efficient large-scale protein production of larvae and pupae of silkworm by Bombyx mori nuclear polyhedrosis virus bacmid system. Biochem Biophys Res Commun 326:564–569 [View Article][PubMed]
    [Google Scholar]
  28. O'Reilly D. R., Miller L. K., Luckow V. A. 1992 Baculovirus Expression Vectors: a Laboratory Manual New York: W.H. Freeman and Company;
    [Google Scholar]
  29. Ono C., Kamagata T., Taka H., Sahara K., Asano S., Bando H. 2012; Phenotypic grouping of 141 BmNPVs lacking viral gene sequences. Virus Res 165:197–206 [View Article][PubMed]
    [Google Scholar]
  30. Partington S., Yu H., Lu A., Carstens E. B. 1990; Isolation of temperature sensitive mutants of Autographa californica nuclear polyhedrosis virus: phenotype characterization of baculovirus mutants defective in very late gene expression. Virology 175:91–102 [View Article][PubMed]
    [Google Scholar]
  31. Post L. E., Palermo D. P., Thomsen D. R., Rottman F. M., Goodwin E. C., Woychik R. P. 1992 Use of a bGH gDNA polyadenylation signal in expression of non-bGH polypeptides in higher eukaryotic cells US patent US5122458 A
    [Google Scholar]
  32. Rohrmann G. F. 2013 Baculovirus Molecular Biology, 2nd edn. Bethesda, MD: National Center for Biotechnology Information (US);
    [Google Scholar]
  33. Rozen S., Skaletsky H. 2000; Primer3 on the WWW for general users and for biologist programmers. Methods Mol Biol 132:365–386[PubMed]
    [Google Scholar]
  34. Ruijter J. M., Ramakers C., Hoogaars W. M., Karlen Y., Bakker O., van den Hoff M. J., Moorman A. F. 2009; Amplification efficiency: linking baseline and bias in the analysis of quantitative PCR data. Nucleic Acids Res 37:e45 [View Article][PubMed]
    [Google Scholar]
  35. Sambrook J., Fritsch E. F., Maniatis T. 1989 Molecular Cloning: a Laboratory Manual, 2nd edn. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory Press;
    [Google Scholar]
  36. Schultz K. L., Friesen P. D. 2009; Baculovirus DNA replication-specific expression factors trigger apoptosis and shutoff of host protein synthesis during infection. J Virol 83:11123–11132 [View Article][PubMed]
    [Google Scholar]
  37. Sharma R. C., Schimke R. T. 1996; Preparation of electrocompetent E. coli using salt-free growth medium. Biotechniques 20:42–44[PubMed]
    [Google Scholar]
  38. Sheng Y., Mancino V., Birren B. 1995; Transformation of Escherichia coli with large DNA molecules by electroporation. Nucleic Acids Res 23:1990–1996 [View Article][PubMed]
    [Google Scholar]
  39. Shikata M., Sano Y., Hashimoto Y., Matsumoto T. 1998; Isolation and characterization of a temperature-sensitive mutant of Bombyx mori nucleopolyhedrovirus for a putative RNA polymerase gene. J Gen Virol 79:2071–2078 [View Article][PubMed]
    [Google Scholar]
  40. Stavroulakis D. A., Kalogerakis N., Behie L. A., Latrou K. 1991; Kinetic data for the BM-5 insect cell line in repeated-batch suspension cultures. Biotechnol Bioeng 38:116–126 [View Article][PubMed]
    [Google Scholar]
  41. Vanarsdall A. L., Okano K., Rohrmann G. F. 2005; Characterization of the replication of a baculovirus mutant lacking the DNA polymerase gene. Virology 331:175–180 [View Article][PubMed]
    [Google Scholar]
  42. Wang L., Yu J., Yin C., Li Z., Hu X., Pang Y. 2002; Characterization of a J domain gene of Spodoptera litura multicapsid nucleopolyhedrovirus. Virus Genes 25:291–297 [View Article][PubMed]
    [Google Scholar]
  43. Wang G.-H., Xia Q.-Y., Cheng D.-J., Duan J., Zhao P., Chen J., Zhu L. 2008; Reference genes identified in the silkworm Bombyx mori during metamorphism based on oligonucleotide microarray and confirmed by qRT-PCR. Insect Sci 15:405–413 [View Article]
    [Google Scholar]
  44. Weiss S. A., Smith G. C., Kalter S. S., Vaughn J. L. 1981; Improved method for the production of insect cell cultures in large volume. In Vitro 17:495–502 [View Article]
    [Google Scholar]
  45. Wilson D., Pethica R., Zhou Y., Talbot C., Vogel C., Madera M., Chothia C., Gough J. 2009; SUPERFAMILY—sophisticated comparative genomics, data mining, visualization and phylogeny. Nucleic Acids Res 37:D380–D386 [View Article][PubMed]
    [Google Scholar]
  46. Xu H. J., Liu Y. H., Yang Z. N., Zhang C. X. 2006; Characterization of ORF39 from Helicoverpa armigera single-nucleocapsid nucleopolyhedrovirus, the gene containing RNA recognition motif. J Biochem Mol Biol 39:263–269 [View Article][PubMed]
    [Google Scholar]
  47. Ye J., Coulouris G., Zaretskaya I., Cutcutache I., Rozen S., Madden T. L. 2012; Primer-BLAST: a tool to design target-specific primers for polymerase chain reaction. BMC Bioinformatics 13:134 [View Article][PubMed]
    [Google Scholar]
  48. Zhang J., Kalogerakis N., Behie L. A., Iatrou K. 1992; Investigation of reduced serum and serum-free media for the cultivation of insect cells (Bm5) and the production of baculovirus (BmNPV). Biotechnol Bioeng 40:1165–1172 [View Article][PubMed]
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jgv/10.1099/jgv.0.000383
Loading
/content/journal/jgv/10.1099/jgv.0.000383
Loading

Data & Media loading...

Supplements

Supplementary Data

PDF
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