Advertisement

Cytotechnology

, Volume 15, Issue 1–3, pp 139–144 | Cite as

Recombinant protein expression in aDrosophila cell line: comparison with the baculovirus system

  • A. R. Bernard
  • T. A. Kost
  • L. Overton
  • C. Cavegn
  • J. Young
  • M. Bertrand
  • Z. Yahia-Cherif
  • C. Chabert
  • A. Mills
Article

Abstract

In this report, we compare two different expression systems: baculovirus/Sf9 and stable recombinantDrosophila Schneider 2 (S2) cell lines. The construction of a recombinant S2 cell line is simple and quick, and in batch fermentations the cells have a doubling time of 20 hours until reaching a plateau density of 20 million cells/ml. Protein expression is driven by theDrosophila Metallothionein promoter which is tightly regulated. When expressed in S2 cells, the extracellular domain of human VCAM, an adhesion molecule, is indistinguishable from the same protein produced by baculovirus-infected Sf9 cells. Additionally, we present data on the expression of a seven trans-membrane protein, the dopamine D4 receptor, which has been successfully expressed in both systems. The receptor integrates correctly in the S2 membrane, binds [3H]spiperone with high affinity and exhibits pharmacological characteristics identical to that of the receptor expressed in Sf9 and mammalian cells. The general implications for large scale production of recombinant proteins are discussed.

Key Words

Baculovirus cell culture Drosophila gene expression insect cell metallothionein promoter recombinant protein 

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. Cavegn C, Young J, Bertrand M and Bernard AR (1994)Drosophila cell lines as hosts for recombinant protein expression. in: Berthold W (ed.) Animal Cell Technology, products for today, prospects for tomorrow. In press. Butterworth-Heineman, Oxford.Google Scholar
  2. Culp JS, Johansen H, Hellmig B, Beck J, Matthews TJ, Delers A and Rosenberg M (1991) Regulated expression allows high level production and secretion of HIV-1 gp 120 envelope glycoprotein inDrosophila Schneider cells. Bio/Technology 9: 173–177.Google Scholar
  3. Johansen H, Van der Straten A, Sweet RI, Otto E, Maroni G and Rosenberg M (1989) Regulated expression at high copy number allows production of a growth inhibitory oncogene product inDrosophila Schneider cells. Genes and Development 3: 882–889.Google Scholar
  4. Maroni G, Otto E and Latowski-Perry D (1986) Molecular and cytogenetic characterization of a metallothionein gene ofDrosophila. Genetics 112: 493–504.Google Scholar
  5. Mills A, Allet B, Bernard A, Chabert C, Brandt E, Cavegn C, Chollet A and Kawashima E (1993) Expression and characterization of human D4 dopamine receptors in baculovirus infected insect cells. FEBS Lett. 320: 130–134.Google Scholar
  6. Mount SM and Rubin GM (1985) Complete nucleotide sequence of theDrosophila transposable element copia: homology between copia and retroviral proteins. Mol. Cell. Biol. 5: 1630–1638.Google Scholar
  7. Olsen MK, Rockenbach SK, Fischer HD, Hoogerheide, G and Tomich CSC (1992) Stable production of an analog of human tissue plasminogen activator from culturedDrosophila cells. Cytotechnology 10: 157–167.Google Scholar
  8. Otto E, Allen JM, Young JE, Palmiter RD and Maroni G (1987) A DNA segment controlling metal-regulated expression ofDrosophila melanogaster Metallothionein gene,Mtn. Mol. Cell. Biol. 7: 1710–1715.Google Scholar
  9. Pavalakis G and Mamer DH (1983) Regulation of a metallthioneingrowth hormone hybrid gene in bovine papilloma virus. Proc. Natl. Acad. Sci. 80: 397–401.Google Scholar
  10. Sang JH (1981)Drosophila cells and cell lines. Adv. Cell Culture 1: 125–177.Google Scholar
  11. Schneider I (1972) Cell lines derived from late embryonic stages ofDrosophila melanogaster. J. Embryol. Exp. Morph. 27 (2): 353–365.Google Scholar
  12. Summers MD and Smith GE (1988) Texas Agricultural Experiment Station Bulletin 1555, Texas A&M University College Station TX USA.Google Scholar
  13. Van Tol HH, Bunzow JR, Guan HC, Sunahara RK, Seeman P, Niznik HB and Civelli O (1991) Cloning of the gene for a human dopamine D4 receptor with high affinity for the antipsychotic clozapine. Nature 350: 610–614.Google Scholar
  14. Van Tol HH, Wu, CM, Guan HC, Ohara K, Bunzow JR, Civelli O, Kennedy J, Seeman P, Niznik HB and Jovanovic V (1992) Multiple dopamine D4 receptor variants in the human population. Nature 358: 149–152.Google Scholar
  15. Walsh GM, Mermod JJ, Hartnell A, Kay AB and Wardlaw AJ (1991) Human eosinophil, but not neutrophil, adherence to IL-lstimulated human umbilical vascular endothelial cells is alpha 4 beta 1 (very late antigen 4) dependent. J. Immunol. 146: 3419–3423.Google Scholar

Copyright information

© Kluwer Academic Publishers 1994

Authors and Affiliations

  • A. R. Bernard
    • 1
  • T. A. Kost
    • 2
  • L. Overton
    • 2
  • C. Cavegn
    • 1
  • J. Young
    • 1
  • M. Bertrand
    • 1
  • Z. Yahia-Cherif
    • 1
  • C. Chabert
    • 1
  • A. Mills
    • 1
  1. 1.Glaxo Institute for Molecular BiologyGenevaSwitzerland
  2. 2.Glaxo Research InstituteResearch Triangle ParkUSA

Personalised recommendations