1887

Abstract

The consequences of locating the polyhedrin gene coding sequences and the p10 promoter at heterologous positions within the nuclear polyhedrosis virus (AcMNPV) genome were investigated. Positioning the polyhedrin or -galactosidase coding sequences under the control of the p10 gene promoter via the use of the new transfer vector, pAcUW 1, resulted in viable recombinant viruses able to produce high levels of each non-fused gene product at the appropriate time. Polyhedra were also produced by the virus with the p10 promoter-polyhedrin hybrid gene and appeared normal in thin sections. Therefore the combination of polyhedrin promoter and coding sequences is evidently not essential for efficient expression of this protein. The p10 promoter can serve this function equally well. Viruses with the p10 promoter and -galactosidase coding sequences placed upstream from the polyhedrin gene in either orientation produced large amounts of -galactosidase protein in infected cells, thus demonstrating that the p10 promoter can function at an alternative position within the virus genome. A second transfer vector, pAc- UW2B, was constructed, with a copy of the p10 gene promoter placed upstream and in opposition to the polyhedrin gene. This mediates the insertion of any foreign gene under the control of the p10 promoter while preserving normal p10 gene expression. The advantages of these constructs over the conventional vectors presently used to express foreign genes in insect cell systems and their utilization in the production of virus insecticides are discussed.

Loading

Article metrics loading...

/content/journal/jgv/10.1099/0022-1317-71-7-1525
1990-07-01
2024-12-07
Loading full text...

Full text loading...

/deliver/fulltext/jgv/71/7/JV0710071525.html?itemId=/content/journal/jgv/10.1099/0022-1317-71-7-1525&mimeType=html&fmt=ahah

References

  1. Bishop D. H. L., Entwistle P. F., Cameron I. R., Allen C. J., Possee R. D. 1988; Field trials with genetically engineered Vaculovirus insecticides. In The Release of Genetically Engineered Micro-organisms pp. 143–179 Sussman M., Collins C. H., Skinner F. A., Stewart-Tull D. E. Edited by London: Academic Press;
    [Google Scholar]
  2. Carstens E. B., Tjia S. T., Doerfler W. 1979; Infection of Spodoptera frugiperda cells with Autographa californica nuclear polyhedrosis virus. Virology 99:386–398
    [Google Scholar]
  3. Cochran M. A., Faulkner P. 1983; Location of homologous DNA sequences interspersed at five regions in the baculovirus AcMNPV genome. Journal of Virology 45:961–970
    [Google Scholar]
  4. Cook R. F., Avery R. J., Dimmock N. J. 1979; Infection of chicken erythrocytes with influenza and other viruses. Infection and Immunity 25:396–402
    [Google Scholar]
  5. Emery V. C., Bishop D. H. L. 1987; The development of multiple expression vectors for high level synthesis of eukaryotic proteins: expression of LCMV N and AcNPV polyhedrin protein by a recombinant baculovirus. Protein Engineering 1:359–366
    [Google Scholar]
  6. Fraser M. J. 1989; Expression of eukaryotic genes in insect cell cultures. In Vitro Cellular and Developmental Biology 25:225–235
    [Google Scholar]
  7. Gonnet P., Devauchelle G. 1987; Obtention par recombination dans le gène du polypeptide plO d’un baculovirus exprimant le gène de resistance à la neomycinedans les cellules d’insects. Comptes rendus de l’Académie des sciences 305:111–114
    [Google Scholar]
  8. Guarino L. A., Summers M.D. 1986; Functional mapping of a trans-activating gene required for expression of a baculovirus delayed-early gene. Journal of Virology 57:563–571
    [Google Scholar]
  9. Howard S. C., Ayres M. D., Possee R. D. 1986; Mapping the 5′ and 3′ ends of Autographa californica polyhedrin mRNA. Virus Research 5:109–119
    [Google Scholar]
  10. Kelly D. C., Lescott T. 1981; Baculovirus replication: protein synthesis in Spodoptera frugiperda cells infected with Trickoplusia ni nuclear polyhedrosis virus. Microbiologica 4:35–57
    [Google Scholar]
  11. Kuzio J., Rohel D. Z., Curry C. J., Krebs A., Carstens E. B., Faulkner P. 1984; Nucleotide sequence of the plO polypeptide gene of Autographa californica nuclear polyhedrosis virus. Virology 139:414–418
    [Google Scholar]
  12. 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]
  13. Luckow V. A., Summers M. D. 1988; Trends in the development of baculovirus expression vectors. Biol Technology 6:47–55
    [Google Scholar]
  14. Maeda S. 1989; Expression of foreign genes in insect cells using baculovirus vectors. Annual Review of Entomology 34:351–372
    [Google Scholar]
  15. Maniatis T., Fritsch E. F., Sambrook J. 1982 Molecular Cloning: A Laboratory Manual New York: Cold Spring Harbor Laboratory;
    [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. Miller L. K. 1988; Baculoviruses as gene expression vectors. Annual Review of Microbiology 42:177–199
    [Google Scholar]
  18. Miller L. K., Trimarchi R. E., Browne D., Pennock G. D. 1983; A temperature-sensitive mutant of the baculovirus Autographa californica nuclear polyhedrosis virus defective in an early function required for further gene expression. Virology 126:376–380
    [Google Scholar]
  19. Pennock G. D., Shoemaker C., Miller L. K. 1984; Strong and regulated expression of Escherichia coli β-galactosidase in insect cells using a baculovirus vector. Molecular and Cellular Biology 4:399–406
    [Google Scholar]
  20. Possee R. D. 1986; Cell-surface expression of influenza virus haemagglutinin in insect cells using a baculovirus vector. Virus Research 5:43–59
    [Google Scholar]
  21. 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]
  22. Qin J., Liu A., Weaver R. F. 1989; Studies on the control region of the plO gene of the Autographa californica nuclear polyhedrosis virus. Journal of General Virology 70:1273–1279
    [Google Scholar]
  23. Rankin C., Ooi B. G., Miller L. K. 1988; Eight base pairs encompassing the transcriptional start point are the major determinants for baculovirus polyhedrin gene expression. Gene 70:39–49
    [Google Scholar]
  24. Rohel D. A., 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]
  25. Rohrmann G. F. 1986; Polyhedrin structure. Journal of General Virology 67:1499–1513
    [Google Scholar]
  26. Smith G. E., Fraser M. J., Summers M. D. 1983a; Molecular engineering of the Autographa californica nuclear polyhedrosis virus genome: deletion mutations within the polyhedrin gene. Journal of Virology 46:584–593
    [Google Scholar]
  27. Smith G. E., Summers M. D., Fraser M. J. 1983b; Production of human β-interferon in insect cells infected with a baculovirus expression vector. Molecular and Cellular Biology 3:2156–2165
    [Google Scholar]
  28. Smith G. E., Vlak J. M., Summers M. D. 1983c; Physical analysis of Autographa californica nuclear polyhedrosis virus transcripts for polyhedrin and 10000-molecular weight protein. Journal of Virology 45:215–225
    [Google Scholar]
  29. Vaughn J. L., Goodwin R. H., Thompkins G. L., Mccawley P. 1977; The establishment of two cell lines from the insect Spodoptera frugiperda (Lepidoptera: Noctuidae). In Vitro 13:213–217
    [Google Scholar]
  30. 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 plO gene of Autographa californica nuclear polyhedrosis virus using a recombinant expressing a p 10-β-galactosidasc fusion gene. Journal of General Virology 69:765–776
    [Google Scholar]
  31. Weyer U., Possee R. D. 1988; Functional analysis of the plO gene 5′ leader sequence of the Autographa californica nuclear polyhedrosis virus. Nucleic Acids Research 16:3635–3653
    [Google Scholar]
  32. Weyer U., Possee R. D. 1989; Analysis of the promoter of the Autographa californica nuclear polyhedrosis virus plO gene. Journal of General Virology 70:203–208
    [Google Scholar]
  33. Williams G. V., Rohel D. Z., Kuzio J., Faulkner P. 1989; A cytopathological investigation of Autographa californica nuclear polyhedrosis virus p10 gene function using insertion/deletion mutants. Journal of General Virology 70:187–202
    [Google Scholar]
  34. Wilson M. E., Mainprize T. H., Friesen P. D., Miller L. K. 1987; Location, transcription and sequence of a baculovirus gene encoding a small arginine-rich polypeptide. Journal of Virology 61:661–666
    [Google Scholar]
/content/journal/jgv/10.1099/0022-1317-71-7-1525
Loading
/content/journal/jgv/10.1099/0022-1317-71-7-1525
Loading

Data & Media loading...

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