Efficient protein production using a nuclear polyhedrosis virus lacking the cysteine proteinase gene Free

Abstract

Infection by a baculovirus ( nuclear polyhedrosis virus, BmNPV) in silkworm () larvae is highly efficient as an expression system for the production of useful proteins. However, the amount of the protein of interest expressed tends to decrease in the later stages of infection presumably due, in part, to a proteinase produced in the larval haemolymph. The N-terminal amino acid sequence of a proteinase purified from the haemolymph of BmNPV-infected larvae was identical to the internal amino acid sequence of the viral cysteine proteinase gene of BmNPV, suggesting that the cysteine proteinase in the haemolymph originated from the BmNPV gene. We constructed a mutant virus (CPd) which had a deletion in the cysteine proteinase gene. No proteinase activity corresponding to this proteinase was detected in the haemolymph of silkworm larvae infected with CPd. The firefly luciferase and the human growth hormone genes were separately introduced into CPd under control of the polyhedrin promoter. These constructs produced these proteins very efficiently, because of a greatly reduced degree of degradation of these proteins. A BmNPV vector system using CPd enhances the stability of foreign expressed proteins, especially for those that are cysteine proteinase-sensitive.

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1997-12-01
2024-03-29
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References

  1. Clem R. J., Fechheimer M., Miller L. K. 1991; Prevention of apoptosis by a baculovirus gene during infection of insect cells. Science 254:1388–1390
    [Google Scholar]
  2. Eguchi M., Iwamoto A. 1976; Alkaline proteases in the midgut tissue and digestive fluid of the silkworm, Bombyx mori. Insect Biochemistry 6:491–496
    [Google Scholar]
  3. Hawtin R. E., Arnold K., Ayres M. D., Zanotto P. M., Howard S. C., Gooday G. W., Chappell L. H., Kitts P. A., King L. A., Possee R. D. 1995; Identification and preliminary characterization of a chitinase gene in the Autographa californica nuclear polyhedrosis virus genome. Virology 212:673–685
    [Google Scholar]
  4. Higashihashi N., Arai Y., Enjo T., Horiuchi T., Saeki Y., Sakano K., Sato Y., Takeda K., Takashina S., Takahashi T. 1991; High-level expression and characterization of hepatitis B virus surface antigen in silkworm using a baculovirus vector. Journal of Virological Methods 35:159–167
    [Google Scholar]
  5. Hill J. E., Kuzio J., Faulkner P. 1995; Identification and characterization of the v-cath gene of the baculovirus, CfMNPV. Biochimica et Biophysica Acta 1264:273–278
    [Google Scholar]
  6. Kadono-Okuda K., Yamamoto M., Higashino Y., Taniai K., Kato Y., Chowdhury S., Xu J., Choi S. K., Sugiyama K., Nakashima K., Maeda S., Yamakawa M. 1995; Baculovirus-mediated production of human growth hormone in the silkworm, Bombyx mori. Biochemical and Biophysical Research Communications 213:389–396
    [Google Scholar]
  7. Kamita S. G., Majima K., Maeda S. 1993; Identification and characterization of the p35 gene of Bombyx mori nuclear polyhedrosis virus that prevents virus-induced apoptosis. Journal of Virology 67:455–463
    [Google Scholar]
  8. Knepper T. P., Arbogast B., Schreurs J., Deinzer M. L. 1992; Determination of the glycosylation patterns, disulfide linkages, and protein heterogeneities of baculovirus-expressed mouse interleukin-3 by mass spectrometry. Biochemistry 31:11651–11659
    [Google Scholar]
  9. Kobayashi M., Mori H., Yaginuma T. 1985; Stimulation of acid protease activity in the isolated pupal abdomens of the silkworm, Bombyx mori, infected with nuclear polyhedrosis virus. Journal of Invertebrate Pathology 46:202–204
    [Google Scholar]
  10. Kobayashi J., Inoue H., Katoh I., Yoshinaka Y., Ikawa Y. 1990; Construction of a new baculovirus vector system of the silkworm, Bombyx mori. in Molecular Insect Science p 325 Hagedorn H. H., Hildebrand J. G., Kidwell M. G., Law J. H. Edited by New York: Plenum;
    [Google Scholar]
  11. Kobayashi J., Imanishi S., Inoue H., Ohsuye K., Yamaichi K., Tsuruoka N., Tanaka S. 1992; High level expression of a frog S- amidating enzyme, AE-II, in cultured cells and silkworm larvae using a Bombyx mori nuclear polyhedrosis virus expression vector. Cytotechnology 8:103–108
    [Google Scholar]
  12. Laemmli U. K. 1970; Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685
    [Google Scholar]
  13. Lebacq-Verheyden A. M., Kasprzyk P. G., Raum M. G., van Wyke Coelingh K., Lebacq J. A., Battey J. F. 1988; Posttranslational processing of endogenous and of baculovirus-expressed human gastrinreleasing peptide precursor. Molecular and Cellular Biology 8:3129–3135
    [Google Scholar]
  14. Maeda S. 1989; Gene transfer vectors of a baculovirus, Bombyx mori, and their use for expression of foreign genes in insect cells. In Invertebrate Cell System Applications pp 167–181 Mitsuhashi J. Edited by Boca Raton, Fla: CRC Press;
    [Google Scholar]
  15. Maeda S. 1994; Expression of foreign genes in insect cells using baculovirus vectors. In Insect Cell Biotechnology pp 1–31 Maramorosch K., McIntosh A. H. Edited by Boca Raton, Fla: CRC Press;
    [Google Scholar]
  16. Maeda S., Kawai T., Obinata M., Fujiwara H., Horiuchi T., Saeki Y., Sato Y., Furusawa M. 1985; Production of human a-interferon in silkworm using a baculovirus vector. Nature 315:592–594
    [Google Scholar]
  17. Murao S., Satoi S. 1970; New pepsin inhibitors (S-PI) from Streptomyces EF-44-201. Agricultural Biological Chemistry 34:1265–1267
    [Google Scholar]
  18. Murao S., Katsura M., Hukuhara K., Oda K. 1980; New metallo proteinase inhibitor (MK-I) produced by Streptomyces mozunensis MK-23. Agricultural Biological Chemistry 44:701–703
    [Google Scholar]
  19. Ohkawa T., Majima K., Maeda S. 1994; A cysteine protease encoded by the baculovirus Bombyx mori nuclear polyhedrosis virus. Journal of Virology 68:6619–6625
    [Google Scholar]
  20. Okazaki H., Kanaya T., Nishimura S., Ogawa K., Watanabe H. 1995; Peroral inoculation of baculovirus vector to the silkworm, Bombyx mori, treated with a low temperature. Journal of Sericultural Science of Japan 64:504–508
    [Google Scholar]
  21. Rawlings N. D., Pearl L. H., Buttle D. F. 1992; The baculovirus Autographa californica nuclear polyhedrosis virus genome includes a papain-like sequence. Biological Chemistry Hoppe-Seyler 373:1211–1215
    [Google Scholar]
  22. Sakurai T., Ueda Y., Sato M., Yanai A. 1992; Feline interferon production in silkworm by recombinant baculovirus. Journal of Veterinary Medical Science 54:563–566
    [Google Scholar]
  23. Slack J. M., Kuzio J., Faulkner P. 1995; Characterization of v-cath, a cathepsin L-like proteinase expressed by the baculovirus Autographa californica multiple nuclear polyhedrosis virus. Journal of General Virology 76:1091–1098
    [Google Scholar]
  24. Suzuki T., Ogawa K., Usami A., Mori H., Matsubara F., Takahashi S., Oda K. 1994; The cysteine proteinase in the hemolymph of silkworm larvae infected with BmNPV. In Program of the Abstracts of the Annual Meeting of the Japanese Society for Sericultural Science Abstract. 523 in Japanese:
    [Google Scholar]
  25. Takahashi S., Ushiyama S., Suzuki T., Ogawa K., Oda K. 1997; Purification and characterization of cysteine proteinase originated in the baculovirus gene. Bioscience Biotechnology and Biochemistry 61:1507–1511
    [Google Scholar]
  26. van Drunen Littel-van den Hurk S., Parker M. D., Fitzpatrick D. R., van den Hurk J. V., Compos M., Babiuk L. A., Zamb T. 1992; Structural, functional, and immunological characterization of bovine herpesvirus-1 glycoprotein gI expressed by recombinant baculovirus. Virology 190:378–392
    [Google Scholar]
  27. Wells D. E., Compans R. W. 1990; Expression and characterization of a functional human immunodeficiency virus envelope glycoprotein in insect cells. Virology 176:575–586
    [Google Scholar]
  28. de Wet J. R., Wood K. V., Deluca M., Helinski D. R., Subramani S. 1987; Firefly luciferase gene: structure and expression in mammalian cells. Molecular and Cellular Biology 7:725–737
    [Google Scholar]
  29. Yamakawa M., Sugisaki K., Morimoto M., Tanaka M., Yamamoto M., Ichikawa T., Nakashima K. 1989; Effects of gene dosage on the expression of human growth hormone cDNA in Escherichia coli. Biochimica et Biophysica Acta 1009:156–160
    [Google Scholar]
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