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

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Cell Culture Engineering IV

Abstract

In this report, we compare two different expression systems: baculovirus/Sf9 and stable recombinant Drosophila 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 the Drosophila Metaliothionein 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.

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References

  • 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 

  • 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 gp120 envelope glycoprotein in Drosophila Schneider cells. Bio/Technology 9: 173–177

    Article  CAS  PubMed  Google Scholar 

  • 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 in Drosophila Schneider cells. Genes and Development 3: 882–889.

    Article  CAS  PubMed  Google Scholar 

  • Maroni G, Otto E and Latowski-Perry D (1986) Molecular and cytogenetic characterization of a metallothionein gene of Drosophila. Genetics 112: 493–504.

    CAS  PubMed  Google Scholar 

  • 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.

    Article  CAS  PubMed  Google Scholar 

  • Mount SM and Rubin GM (1985) Complete nucleotide sequence of the Drosophila transposable element copia: homology between copia and retroviral proteins. Mol. Cell. Biol. 5: 1630–1638

    CAS  PubMed Central  PubMed  Google Scholar 

  • Olsen MK, Rockenbach SK, Fischer HD, Hoogerheide G and Tomich CSC (1992) Stable production of an analog of human tissue plasminogen activator from cultured Drosophila cells. Cytotechnology 10: 157–167.

    Article  CAS  PubMed  Google Scholar 

  • Otto E, Allen JM, Young JE, Palmiter RD and Maroni G (1987) A DNA segment controlling metal-regulated expression of Drosophila melanogaster Metallothionein gene Mtn. Mol. Cell. Biol. 7: 1710–1715.

    CAS  PubMed Central  PubMed  Google Scholar 

  • Pavalakis G and Mamer DH (1983) Regulation of a metallthionein-growth hormone hybrid gene in bovine papilloma virus. Proc. Natl.Acad. Sci. 80: 397–401.

    Article  Google Scholar 

  • Sang JH (1981) Drosophila cells and cell lines. Adv. Cell Culture 1: 125–177.

    CAS  Google Scholar 

  • Schneider I (1972) Cell lines derived from late embryonic stages of Drosophila melanogaster. J. Embryol. Exp. Morph. 27(2): 353–365.

    CAS  PubMed  Google Scholar 

  • Summers MD and Smith GE (1988) Texas Agricultural Experiment Station Bulletin 1555, Texas A&M University College Station TX USA.

    Google Scholar 

  • Van Toi 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.

    Article  Google Scholar 

  • 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.

    Article  PubMed  Google Scholar 

  • Walsh GM, Mermod JJ, Hartnell A, Kay AB and Wardlaw AJ (1991) Human eosinophil, but not neutrophil, adherence to IL-1-stimulated human umbilical vascular endothelial cells is alpha 4 beta 1 (very late antigen-4) dependent. J. Immunol. 146: 3419–3423.

    CAS  PubMed  Google Scholar 

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© 1994 Springer Science+Business Media Dordrecht

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Bernard, A.R. et al. (1994). Recombinant protein expression in a Drosophila cell line: comparison with the baculovirus system. In: Buckland, B.C., Aunins, J.G., Bibila, T.A., Hu, WS., Robinson, D.K., Zhou, W. (eds) Cell Culture Engineering IV. Current Applications of Cell Culture Engineering, vol 1. Springer, Dordrecht. https://doi.org/10.1007/978-94-011-0257-5_16

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  • DOI: https://doi.org/10.1007/978-94-011-0257-5_16

  • Publisher Name: Springer, Dordrecht

  • Print ISBN: 978-94-010-4114-0

  • Online ISBN: 978-94-011-0257-5

  • eBook Packages: Springer Book Archive

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