1887

Abstract

We have evaluated the use of baculoviruses to deliver polydnavirus (CsPDV) genomic DNA into lepidopteran larvae to facilitate the identification of functional CsPDV genes. Genomic fragments consisting of regulatory (promoter) and coding sequences for two CsPDV genes (VHv1.1 and WHv1.6) were used to generate CsPDV-baculovirus recombinants and evaluate the expression of genes under the regulation of the CsPDV promoters. Northern blot and primer extension studies established that CsPDV genes were expressed under the control of their own promoters in these CsPDV-baculovirus recombinants. Transcripts were detected as early as 4 h post-infection indicating that temporal activity of CsPDV promoters was retained. The VHv1.1 gene product as expressed from CsPDV-baculovirus recombinants was identical in size and in functional properties to that produced in CsPDV-infected insects. CsPDV-baculovirus recombinants may be useful for the screening and characterization of polydnavirus genes with functional activities that can only be evaluated in insect larvae.

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1996-07-01
2022-01-20
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References

  1. Blissard G. W., Rohrmann G. F. 1990; Baculovirus diversity and molecular biology. Annual Review of Entomology 35:127–155
    [Google Scholar]
  2. Blissard G. W., Vinson S. B., Summers M. D. 1986; Identification, mapping, and in vitro translation of Campoletis sonorensis virus mRNAs from parasitized Heliothis virescens larvae. Journal of Virology 57:318–327
    [Google Scholar]
  3. Blissard G. W., Smith O. P., Summers M. D. 1987; Two related viral genes are located on a single superhelical DNA segment of the multipartite Campoletis sonorensis virus genome. Virology 160:120–134
    [Google Scholar]
  4. Blissard G. W., Theilmann D. A., Summers M. D. 1989; Segment W of Campoletis sonorensis virus: expression, gene products, and organization. Virology 169:78–89
    [Google Scholar]
  5. Chomczynski P., Sacchi N. 1987; Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Analytical Biochemistry 162:156–159
    [Google Scholar]
  6. Cui L., Webb B. A. 1996; Isolation and characterization of a member of the cysteine-rich gene family from Campoletis sonorensis polydnavirus. Journal of General Virology 77:797–809
    [Google Scholar]
  7. Dib-Hajj S. D., Webb B. A., Summers M. D. 1993; Structure and evolutionary implications of a ‘cysteine-rich’ Campoletis sonorensis polydnavirus gene family. Proceedings of the National Academy of Sciences, USA 90:3765–3769
    [Google Scholar]
  8. Fathpour H., Dahlman D. L. 1995; Polydnavirus of Micropliiis croceipes prolongs the larval period and changes hemolymph protein content of the host, Heliothis virescens. Archives of Insect Biochemistry and Physiology 28:33–48
    [Google Scholar]
  9. Fleming J. G. W., Summers M. D. 1990; The integration of the genome of a segmented DNA virus in the host insect’s genome. In Molecular Insect Science pp 99–105 Edited by Hagedorn H. H. New York: Plenum Press;
    [Google Scholar]
  10. Granados R. R., Naughton M. 1976; Replication of Amsacta moorei entomopoxvirus in ovarian and hemocyte cultures from Estigmene acrea larvae. In Invertebrate Tissue Culture, Applications in Medicine, Biology, and Agriculture pp 472–476 Edited by Kurstak E., Maramorosch K. New York: Academic Press;
    [Google Scholar]
  11. Latrou K., Meidinger R. G. 1990; Tissue-specific expression of silkmoth chorion genes in vivo using Bombyx mori nuclear polyhedrosis virus as a transducing vector. Proceedings of the National Academy of Sciences, USA 87:3650–3654
    [Google Scholar]
  12. King L. A., Possee R. D. 1992 The Baculovirus Expression System. A Laboratory Guide London: Chapman and Hall;
    [Google Scholar]
  13. Kreil P. J., Summers M. D., Vinson S. B. 1982; Virus with a multipartite superhelical DNA genome from the ichneumonid parasitoid Campoletis sonorensis. Journal of Virology 43:859–870
    [Google Scholar]
  14. Li X., Webb B. A. 1994; Apparent functional role for a cysteine-rich polydnavirus protein in suppression of the insect cellular immune response. Journal of Virology 68:7482–7489
    [Google Scholar]
  15. Lingren P. D., Guerra R. J., Nickelsen J. W., White C. 1970; Hosts and host-age preference of Campoletis perdistinctus. Journal of Economic Entomology 63:518–522
    [Google Scholar]
  16. O’Reilly D. R., Miller L. K., Luckow V. A. 1992 Baculovirus Expression Vectors: A Laboratory Manual New York: W. H. Freeman;
    [Google Scholar]
  17. Sambrook J., Fritsch E., Maniatis T. 1989 Molecular Cloning: A Laboratory Manual NY: Cold Spring Harbor Laboratory;
    [Google Scholar]
  18. Shelby K. S., Webb B. A. 1994; Polydnavirus infection inhibits synthesis of an insect plasma protein, arylphorin. Journal of General Virology 75:2285–2292
    [Google Scholar]
  19. Strand M. R., Pech L. L. 1995; Microplitis demolitor polydnavirus induces apoptosis of a specific haemocyte morphotype in Pseudoplusia includens. Journal of General Virology 76:283–291
    [Google Scholar]
  20. Summers M. D., Dib-Hajj S. D. 1995; Polydnavirus-facilitated endoparasite protection against host immune defenses. Proceedings of the National Academy of Sciences, USA 92:29–36
    [Google Scholar]
  21. Theilmann D. A., Stewart S. 1992; Molecular analysis of the transactivating IE-2 gene of Orgyia pseudotsugata multicapsid nuclear polyhedrosis virus. Virology 187:84–96
    [Google Scholar]
  22. Webb N. R., Summers M. D. 1990; Expression of proteins using recombinant baculovirus. Techniques-A Journal of Methods in Cell and Molecular Biology 2:173–188
    [Google Scholar]
  23. Wickman T. J., Nemerow G. R. 1993; Optimization of growth methods and recombinant protein production in BTI-Tn-5B1-4 insect cells using the baculovirus expression system. Biotechnology Progress 9:25–30
    [Google Scholar]
  24. Wickman T. J., Davis T., Granados G. R., Shuler M. L., Wood H. A. 1992; Screening of insect cell lines for the production of recombinant proteins and infectious virus in the baculovirus expression system. Biotechnology Progress 8:391–396
    [Google Scholar]
  25. Xu D., Stoltz D. 1993; Polydnavirus genome segment families in the ichneumonid parasitoid Hyposoter fugitivus. Journal of Virology 67:1340–1349
    [Google Scholar]
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