1887

Abstract

An improved polyhedrin-based baculovirus expression vector was constructed to expedite distinguishing infections by putative baculovirus recombinants from infections by wild-type (wt) baculovirus. The vector utilizes the -galactosidase gene () as a genetic marker for positive recombination between wt nuclear polyhedrosis virus and the baculovirus transfer vector. The marker gene/expression cassette was constructed so that and the deleted polyhedrin gene were transcribed in opposite orientations, both terminating in a simian virus 40 DNA fragment which acts as a bidirectional terminator. In the constructed vector, is transcribed from the heat-shock promoter (hsp70), which is constitutively expressed in baculovirus-infected (Sf) cells, thereby making the site of the deleted polyhedrin gene available for the insertion and expression of foreign genes under the control of the polyhedrin promoter. Recombinant baculoviruses are readily selected in plaque assays by the development of a blue colour upon the addition of X-Gal. The colour selection renders the retrieval of recombinants less dependent on a high frequency of recombination between the transfer vector and wt baculovirus DNA. The usefulness of this new vector was illustrated by expressing gene I of cauliflower mosaic virus, which encodes a protein of 46000. Expression of gene I was at the same level as in cells infected with a conventional polyhedrin-based expression vector. Gene I protein formed large hollow fibre-like structures in the cytoplasm of infected Sf cells. This is the first plant virus protein to be expressed in insect cells by a recombinant baculovirus.

Loading

Article metrics loading...

/content/journal/jgv/10.1099/0022-1317-71-10-2201
1990-10-01
2022-01-19
Loading full text...

Full text loading...

/deliver/fulltext/jgv/71/10/JV0710102201.html?itemId=/content/journal/jgv/10.1099/0022-1317-71-10-2201&mimeType=html&fmt=ahah

References

  1. Albrecht H., Geldreich A., Menissier De Murcia J., Kirchherr D., Mesnard J. -M., Lebeurier G. 1988; Cauliflower mosaic virus gene I product detected in a cell-wall-enriched fraction. Virology 163:503–508
    [Google Scholar]
  2. Capone J. 1989; Screening recombinant baculovirus plaques in situ with antibody probes. Gene Analytical Techniques 6:62–66
    [Google Scholar]
  3. Casadaban M. J. J., Martinez-Arias A., Shapira S. K., Chou J. 1981; Beta-galactosidase gene fusions for analyzing gene expression in Escherichia coli and yeast. Methods in Enzymology 100:293–308
    [Google Scholar]
  4. Cochran M. A., Carstens E. B., Eaton B. E., Faulkner P. 1982; Molecular cloning and physical mapping of restriction endonuclease fragments of Autographa californica nuclear poly-hedrosis virus. Journal of Virology 41:940–946
    [Google Scholar]
  5. Covey S. N. 1985; Organisation and expression of the cauliflower mosaic virus genome. In Molecular Plant Virology, 2 pp 121–159 Davies J. W. Edited by Boca Raton: CRC Press;
    [Google Scholar]
  6. 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]
  7. Franck A., Guilley H., Jonard G., Richards K., Hirth L. 1980; Nucleotide sequence of cauliflower mosaic virus DNA. Cell 21:285–294
    [Google Scholar]
  8. Granados R. R., Federici B. A. 1986 editor . The Biology of Baculoviruses 2 Practical Applications for Insect Control Boca Raton; CRC Press:
    [Google Scholar]
  9. Harker C. L., Mullineaux P. M., Bryant J. A., Maule A. J. 1987; Detection of CaMV gene I and gene VI protein products in vivo using antisera raised to COOH-terminal β-galactosidase fusions proteins. Plant Molecular Biology 8:275–287
    [Google Scholar]
  10. Hink F. 1970; Established insect cell line from the cabbage looper, Trichoplusia ni. Nature; London: 226466–467
    [Google Scholar]
  11. Kalderon D., Roberts B. L., Richardson W. D., Smith A. E. 1984; A short amino acid sequence able to specify nuclear location. Cell 39:499–509
    [Google Scholar]
  12. Karch F., Török I., Tissière A. 1981; Extensive regions of homology in front of the hsp70 heat shock variant genes in Drosophila melanogaster. Journal of Molecular Biology 148:219–230
    [Google Scholar]
  13. Kelly D. C. 1982; Baculovirus replication. Journal of General Virology 63:1–13
    [Google Scholar]
  14. Kunkel T. A., Roberts J. D., Zakour R. A. 1987; Rapid and efficient site-specific mutagenesis without phenotypic selection. Methods in Enzymology 154:367–382
    [Google Scholar]
  15. Laemmli U. K. 1970; Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature; London: 227680–685
    [Google Scholar]
  16. Linstead P. J., Hills G. J., Plaskitt K. A., Wilson I. G., Harker C. L., Maule A. J. 1988; The subcellular location of the gene 1 product of cauliflower mosaic virus is consistent with a function associated with virus spread. Journal of General Virology 69:1809–1818
    [Google Scholar]
  17. Luckow V. L., Summers M. D. 1988; Trends in the development of baculovirus expression vectors. Bio/Technology 6:47–55
    [Google Scholar]
  18. Maeda S. 1989; Expression of foreign genes in insects using baculovirus vectors. Annual Review of Entomology 34:351–372
    [Google Scholar]
  19. Maniatis T., Fritsch E. F., Sambrook J. 1982 Molecular Cloning: A Laboratory Manual New York: Cold Spring Harbor Laboratory;
    [Google Scholar]
  20. Matthews R. E. F. 1982; Classification and nomenclature of viruses. Intervirology 12:129–296
    [Google Scholar]
  21. Maule A. J. 1985; Replication of caulimoviruses in plants and protoplasts. In Molecular Plant Virology 2 pp 161–190 Davies J. W. Edited by Davies Boca Raton: CRC Press;
    [Google Scholar]
  22. Maule A. J., Harker C. L., Wilson I. G. 1989; The pattern of accumulation of cauliflower mosaic virus-specific products in infected turnips. Virology 169:436–446
    [Google Scholar]
  23. Miller L. K. 1988; Baculoviruses as gene expression vectors. Annual Review of Microbiology 42:177–199
    [Google Scholar]
  24. Miller L. K. 1989; Insect baculoviruses: powerful gene expression vectors. Bio Assays 11:91–95
    [Google Scholar]
  25. Pen J., Welling G. W., Welling-Wester S. 1989; An efficient procedure for the isolation of recombinant baculovirus. Nucleic Acids Research 17:451
    [Google Scholar]
  26. 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]
  27. Sambrook J., Fritsch E. F., Maniatis T. 1989; . Molecular Cloning: A Laboratory Manual, 2nd edn.. New York: Cold Spring Harbor Laboratory;
    [Google Scholar]
  28. Smith G. E., Summers M. D. 1978; Analysis of baculovirus genomes with restriction endonucleases. Virology 89:517–527
    [Google Scholar]
  29. Smith G. E., Fraser M. J., Summers M. D. 1983; Molecular engineering of the Autographa californica nuclear polyhedrosis virus genome: deletion mutations within the polyhedrin gene. Journal of Virology 46:584–593
    [Google Scholar]
  30. Summers M. D., Smith G. E. 1987; A manual of methods for baculovirus vectors and insect cell culture procedures. Texas Agricultural Experimental Station Bulletin 1555
    [Google Scholar]
  31. Van Der Wilk F., Vanlent J. W. M., Vlak J. M. 1987; Immunogold detection of polyhedrin, p10 and virion antigens in Autographa californica nuclear polyhedrosis virus-infected Spodoptera frugiperda. Journal of General Virology 68:2615–2623
    [Google Scholar]
  32. Van Lent J. W. M., Groenen J. T. M., Klinge-Roode E. C., Rohrmann G. F., Zuidema D., Vlak J. M. 1990; Localization of the 36-kDa polyhedron envelope protein in Spodoptera frugiperda cells infected with Autographa californica nuclear polyhedrosis virus. Archives of Virology 111:103–114
    [Google Scholar]
  33. Vaughn J. L., Goodwin R. H., Tompkins G. J., Mccawley P. 1977; The establishment of two cell lines from the insect (Lepidoptera: Noctuidae). In Vitro 13:213–217
    [Google Scholar]
  34. Vialard J., Lalumiere M., Vernet T., Briedis D., Alkhatib G., Henning D., Levin D., Richardson C. 1990; Synthesis of the membrane fusion and hemagglutinin proteins of measles virus using a novel baculovirus vector containing the β-galactosidase gene. Journal of Virology 64:37–50
    [Google Scholar]
  35. Vlak J. M., Odink K. G. 1979; Characterization of Autographa californica nuclear polyhedrosis virus deoxyribonucleic acid. Journal of General Virology 44:333–347
    [Google Scholar]
  36. Vlak J. M., Klinkenberg F. A., Zaal K. I. M., Usmany M., Klinge-Roode E. C., Geervliet I. B. F., Roosien J., Vanlent 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]
  37. Vlak J. M., Schouten A. J., Usmany M., Belsham G. J., Klinge-Roode E. C., Maule A. J., Vanlent J. W. M., Zuidema D. 1990; Expression of cauliflower mosaic virus gene I using a baculovirus vector based upon the p10 gene and a novel selection method. Virology in press
    [Google Scholar]
  38. 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]
http://instance.metastore.ingenta.com/content/journal/jgv/10.1099/0022-1317-71-10-2201
Loading
/content/journal/jgv/10.1099/0022-1317-71-10-2201
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