1887

Abstract

Summary

5′ deletion mutants of the nuclear polyhedrosis virus very late p 10 gene promoter have been prepared and subjected to a transient expression assay in infected cells. The control plasmid contained the chloramphenicol acetyltransferase (CAT) reporter gene under the control of the p 10 promoter, which was included in a 230 bp sequence upstream from the p 10 translation initiation codon. The control plasmid also contained a segment of the hr enhancer downstream from the CAT gene. Promoter activity was unaffected by 5′ deletion to position –77, which lies about 11 bp upstream from the p10 cap site. However, deletion of 12 more bp completely eliminated p10 promoter activity. Thus, the 5′ border of the p10 promoter lies downstream from position –77, and the region between positions –77 and –65 contains an element that is important to promoter activity. This is the region that is conserved near the cap sites of late baculovirus genes. Our studies also show that transient expression of CAT under the control of the p10 promoter and hr enhancer is higher when transfection occurs prior to infection by virus.

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1989-05-01
2024-04-25
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References

  1. Birnboim H. C., Doly J. 1979; A rapid alkaline extraction procedure for screening recombinant plasmid DNA. Nucleic Acids Research 1:1513–1523
    [Google Scholar]
  2. Chen E. Y., Seeburg P. H. 1985; Supercoil sequence: a fast simple method for sequencing plasmid DNA. DNA 4:165–170
    [Google Scholar]
  3. Fuchs L. Y., Woods M. S., Weaver R. F. 1983; Viral transcription during Autographa californica nuclear polyhedrosis virus infection: a novel RNA polymerase induced in infected Spodoptera frugiperda cells. Journal of Virology 48:641–646
    [Google Scholar]
  4. Garger S. J., Griffith O. M., Grill L. K. 1983; Rapid purification of plasmid DNA by a single centrifugation in a two-step cesium chloride-ethidium gradient. Biochemical and Biophysical Research Communications 117:835–842
    [Google Scholar]
  5. Gorman C. M., Moffat L. F., Howard B. H. 1982; Recombinant genomes which express chloramphenicol acetyltransferase in mammalian cells. Molecular and Cellular Biology 2:1044–1051
    [Google Scholar]
  6. Graham F. L., Van Der Eb A. J. 1973; A new technique for the assay of infectivity of human adenovirus 5 DNA. Virology 52:456–467
    [Google Scholar]
  7. Grula M., Buller P. L., Weaver R. F. 1981; a-amanitin-resistant viral RNA synthesis in nuclei isolated from nuclear polyhedrosis virus-infected Heliothis zea larvae and Spodoptera frugiperda cells. Journal of Virology 38:916–921
    [Google Scholar]
  8. Guarino L. A., Summers M. D. 1986; Interspersed homologous DNA of Autographa californica nuclear polyhedrosis virus enhances delayed-early gene expression. Journal of Virology 60:215–223
    [Google Scholar]
  9. Guo L. H., Wu R. 1983; Exonuclease III: use for DNA sequence analysis and in specific deletions of nucleotides. Methods in Enzymology 100:60–96
    [Google Scholar]
  10. Knebel D., Doerfler W. 1987; Activation of an insect baculovirus promoter in mammalian cells by adenovirus functions. Virus Research 8:317–326
    [Google Scholar]
  11. Kuzio J., Rohel D. Z., Curry C. J., Krebs A., Carstens E. B., Faulkner P. 1984; Nucleotide sequence of the p10 polypeptide gene of Autographa californica nuclear polyhedrosis virus. Virology 139:414–418
    [Google Scholar]
  12. Lee H. H., Miller L. K. 1978; Isolation of genotypic variants of Autographa californica nuclear polyhedrosis virus. Journal of Virology 27:754–767
    [Google Scholar]
  13. Liu A., Qin J., Rankin C, Hardin S. E., Weaver R. F. 1986; Nucleotide sequence of a portion of the Autographa californica nuclear polyhedrosis virus genome containing the EcoRI site-rich region (hr5) and an open reading frame just 5′ of the p10 gene. Journal of General Virology 67:2565–2570
    [Google Scholar]
  14. Lübbert H., Doerfler W. 1984; Transcription of overlapping sets of RNAs from the genome of Autographa californica nuclear polyhedrosis virus: a novel method for mapping RNAs. Journal of Virology 52:255–265
    [Google Scholar]
  15. Masters B. S., Stohl L. L., Clayton D. A. 1987; Yeast mitochondrial RNA polymerase is homologous to those encoded by bacteriophages T3 and T7. Cell 51:89–99
    [Google Scholar]
  16. Matsuura Y., Possee R. D., Overton H. A., Bishop D. H. L. 1987; Baculovirus expression vectors: the requirements for high level expression of proteins, including glycoproteins. Journal of General Virology 68:1233–1250
    [Google Scholar]
  17. Possee R. D., Howard S. C. 1987; Analysis of the polyhedrin gene promoter of the Autographa californica nuclear polyhedrosis virus. Nucleic Acids Research 15:10233–10248
    [Google Scholar]
  18. Rankin C, Ladin B. F., Weaver R. F. 1986; Physical mapping of temporally regulated, overlapping transcripts in the region of the 10K protein in Autographa californica nuclear polyhedrosis virus. Journal of Virology 57:18–27
    [Google Scholar]
  19. Rohel D. Z., Cochran M. A., Faulkner P. 1983; Characterization of two abundant mRNAs of Autographa californica nuclear polyhedrosis virus present late in infection. Virology 124:357–365
    [Google Scholar]
  20. Rohrmann G. F. 1986; Polyhedrin structure. Journal of General Virology 67:1499–1513
    [Google Scholar]
  21. Sanger F., Nicklen S., Coulson A. R. 1977; DNA sequencing with chain-terminating inhibitors. Proceedings of the National Academy of SciencesU.S.A 74:5463–5467
    [Google Scholar]
  22. Smith G. E., Vlak J. M., Summers M. D. 1982; In vitro translation of Autographa californica nuclear polyhedrosis virus early and late mRNAs. Journal of Virology 44:199–208
    [Google Scholar]
  23. 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. Journal of Virology 45:215–225
    [Google Scholar]
  24. Vaughn J. L., Goodwin R. H., Thompkins G. L., McCawly P. 1977; Establishment of two insect cell lines from the insect Spodoptera frugiperda (Lepidoptera: Noctuidae). In Vitro 13:213–217
    [Google Scholar]
  25. Vlak J. M., Klinkenberg F. A., Zaal K. J. M., Usmany M., Klinge-Roode E. C., Geervliet J. B. F., Roosien J., Van Lent J. W. M. 1988; Functional studies on the p10 gene of Autographa californica nuclear polyhedrosis virus using a recombinant expressing a pl0-β-galactosidase fusion gene. Journal of General Virology 69:765–776
    [Google Scholar]
  26. Weyer U., Possee R. D. 1988; Functional analysis of the p10 gene 5′ leader sequence of the Autographa californica nuclear polyhedrosis virus. Nucleic Acids Research 16:3635–3653
    [Google Scholar]
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