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Strategies for Expressing Cloned Genes in Mammalian Cells

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Protein Production by Biotechnology

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Abstract

The ability to express cloned genes in mammalian cell lines has proved to be a powerful technology. It is now possible to produce a rare protein of some scientific or therapeutic interest in sufficient quantity to be able to study it and/or produce it commercially. With the genes coding for interesting proteins cloned and expressable, the future holds the possibility for genetically modifying natural proteins to design and produce new proteins as required. The best example, to date, of a cloned gene expressed in mammalian cells to produce a human therapeutic agent is tissue plasminogen activator (tPA). Interestingly, there are already underway projects to engineer natural tPA protein to create new ‘second-generation’ tPA proteins with altered properties such as longer half-life and increased substrate specificity1 (see Chapter 8, this volume).

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References

  1. Schnee, J. M., Runge, M. S., Matsueda, G. R., Hudson, N. W., Seidman, J. G., Haber, E. & Quertermous, T., Construction and expression of a recombinant antibody-targeted plasminogen activator. Proc. Nat. Acad. Sci., USA, 84 (1987) 6904–8.

    Article  CAS  Google Scholar 

  2. Harris, T. J. R., Expression of eukaryotic genes in E. coli. In Genetic Engineering, Vol. 4, ed. R Williamson, Academic Press, London, 1983, pp. 127–85.

    Google Scholar 

  3. Bebbington, C. R. & Hentschel, C. C., The use of vectors based on gene-amplification for the expression of cloned genes in mammalian cells. In DNA Cloning, Vol. III, ed. D. M. Glover. IRL Press, Oxford, 1987, pp. 163–88.

    Google Scholar 

  4. Kaufman, R J. & Sharp, P. A., Amplification and expression of sequences cotransfected with a modular dihydrofolate reductase complementary DNA gene. J. Molec. Biol., 159 (1982) 601–21.

    Article  PubMed  CAS  Google Scholar 

  5. Stephens, P. E. & Hentschel, C. C., The bovine papilloma virus genome and its uses as a eukaryotic vector. Biochem. J., 248 (1987) 1–11.

    PubMed  CAS  Google Scholar 

  6. Mulligan, R C. & Berg, P., Selection for animal cells that express the Escherichia coli gene coding for xanthine-guanine phosphoribosyltransferase. Proc. Nat. Acad. Sci., USA, 78, 2072–6.

    Google Scholar 

  7. Morrison, S. L., Transfectomas provide novel chimeric antibodies. Science, 229 (1985) 1202–7.

    Article  PubMed  CAS  Google Scholar 

  8. Swartz, R., Selecting a preparative mammalian cell production technology in 1988. Gen. Engng News, 8 (1988) 14–35.

    Google Scholar 

  9. Schimke, R T., Gene amplification in cultured animal cells. Cell, 37 (1984) 705–13.

    Article  PubMed  CAS  Google Scholar 

  10. Stark, G. R. & Wahl, G. M., Gene amplification. Ann. Rev. Biochem., 53 (1984) 447–91.

    Article  PubMed  CAS  Google Scholar 

  11. Stark, G. R., DNA amplification in drug resistant cells and in tumours. Cancer Surveys, 5 (1986) 1–23.

    PubMed  CAS  Google Scholar 

  12. Kaufman, R J., Murtha, P., Ingolia, D. E., Yeung, C. Y. & Kellens, R E., Selection and amplification of heterologous genes encoding adenosine deaminase in mammalian cells. Proc. Nat. Acad. Sci., USA, 83 (1986) 3136–40.

    Article  CAS  Google Scholar 

  13. Christman, J. K., Gerber, M., Price, P. M., Flordellis, C., Edelman, J. & Acs, G., Amplification of expression of hepatitis B surface anitgen in 3T3 cells cotransfected with a dominant-acting gene and cloned viral DNA. Proc. Nat. Acad. Sci., USA, 79 (1982) 1815–19.

    Article  CAS  Google Scholar 

  14. Murray, M. J., Kaufman, R. J., Latt, S. A. & Weinberg, R. A., Construction and use of a dominant, selectable marker: a Harvey sarcoma virus-dihydrofolate reductase chimera. Molec. Cell Biol., 3 (1983) 32–43.

    PubMed  CAS  Google Scholar 

  15. Deschatrette, J., Fougere-Deschatrette, C., Corcos, L. & Schimke, R T., Expression of the mouse serum albumin gene introduced into differentiated and dedifferentiated rat hepatoma cells. Proc. Nat. Acad. Sci., USA, 82 (1985) 765–9.

    Article  CAS  Google Scholar 

  16. Michel, M. L., Sobczak, E., Malpiece, Y., Tiollais, P. & Streeck, R. E., Expression of amplified hepatitis B virus surface antigen genes in Chinese hamster ovary cells. Bio/Technology, 3 (1985) 561–6.

    Article  CAS  Google Scholar 

  17. Kaufman, R. J., Wasley, L. C., Spiliotes, A. J., Gossels, S. D., Latt, S. A., Larsen, G. R. & Kay, R. M., Coamplification and coexpression of human tissue-type plasminogen activator and murine dihydrofolate reductase sequences in Chinese hamster ovary cells. Molec. Cell. Biol., 5 (1985) 1750–9.

    PubMed  CAS  Google Scholar 

  18. Mory, Y., Ben-Barak, J., Seger, D., Cohen, B., Novick, D., Fischer, D. G., Rubinstein, M., Kargman, S., Zilberstein, A., Vigneron, M. & Revel, M., Efficient constitutive production of human IFN-γ in Chinese hamster ovary cells. DNA, 5 (1986) 181–93.

    Article  PubMed  CAS  Google Scholar 

  19. Patzer, E. J., Nakamura, G. R., Hershberg, R. D., Gregory, T. J., Crowley, C., Levinson, A. D. & Eichberg, J. W., Cell culture derived recombinant HBsAg is highly immunogenic and protects chimpanzees from infection with hepatitis B virus. Bio/Technology, 4 (1986) 630–6.

    Article  CAS  Google Scholar 

  20. DiMaio, D., Papillomavirus cloning vectors. In The Papillomaviruses, Plenum Press, New York, 1984.

    Google Scholar 

  21. Lowy, D. R., Rands, E. & Scolnick, E. M., Helper-independent transformation by unintegrated Harvey sarcoma virus DNA. J. Virol., 26 (1978) 291–8.

    PubMed  CAS  Google Scholar 

  22. Lowy, D. R., Dvoretzky, I., Shober, R., Law, M. F., Engel, L. & Howley, P. M., In vitro transformation by a defined subgenomic fragment of bovine papilloma virus DNA. Nature, 287 (1980) 72–4.

    Article  PubMed  CAS  Google Scholar 

  23. Campo, M. S., Spandidos, D. A., Lang, J. and Wilkie, N. M., Transcriptional control signals in the genome of bovine papillomavirus type 1. Nature, 303 (1983) 77–80.

    Article  PubMed  CAS  Google Scholar 

  24. Sambrook, J., Rodgers, L., White, J. & Gething, M. J., Lines of BPV-transformed murine cells that constitutively express influenza virus hemagglutinin. EMBO J., 4 (1985) 91–103.

    PubMed  CAS  Google Scholar 

  25. Pavlakis, G. N. & Hamer, D. H., Regulation of a metallothionein-growth hormone hybrid gene in bovine papilloma virus. Proc. Nat. Acad. Sci., USA, 80 (1983) 397.

    Article  CAS  Google Scholar 

  26. Hsiung, N., Fitts, R., Wilson, S., Milne, A. & Hamer, D., Efficient production of hepatitis B surface antigen using a bovine papilloma virus-metallothienein vector. J. Molec. Appl. Genet., 2 (1984) 497–506.

    CAS  Google Scholar 

  27. Stephens, P. E., Bendig, M. M. & Hentschel, C. C., Expression of human tissue plasminogen activator in mouse cells. J. Cell Biochem., Suppl. 9C (1985) 75.

    Google Scholar 

  28. Bendig, M. M., Stephens, P. E., Cockett, M. I. & Hentschel, C. C., Mouse cell lines that use heat shock promoters to regulate the expression of tissue plasminogen activator. DNA, 6 (1987) 343–52.

    Article  PubMed  CAS  Google Scholar 

  29. Fukunaga, R., Sokawa, Y. & Nagata, S., Constitutive production of human interferons by mouse cells with bovine papillomavirus as a vector. Proc. Nat. Acad. Sci., USA, 81 (1984) 5086–90.

    Article  CAS  Google Scholar 

  30. Zinn, K., Mellon, P., Ptashne, M. & Maniatis, T., Regulated expression of an extrachromosomal human β-interferon gene in mouse cells. Proc. Nat. Acad. Sci., USA, 79 (1982) 4897–901.

    Article  CAS  Google Scholar 

  31. Mitrani-Rosenbaum, S., Maroteaux, L., Mory, Y., Revel, M. & Howley, P. M., Inducible expression of the human interferon 1 gene linked to a bovine papilloma virus DNA vector and maintained extrachromosomally in mouse cells. Molec. Cell Biol., 3 (1983) 233–240.

    PubMed  CAS  Google Scholar 

  32. de St. Groth, S. F., Automated production of monoclonal antibodies in a cytostat. J. Immunol. Methods, 57 (1983) 121–36.

    Article  Google Scholar 

  33. Birch, J. R., Boraston, R. & Wood, L., Bulk production of monoclonal antibodies in fermenters. Trends Biotechnol, 3 (1985) 162–6.

    Article  Google Scholar 

  34. Weidle, U. H. & Buckel, P., Establishment of stable mouse myeloma cells constitutively secreting human tissue-type plasminogen activator. Gene, 57 (1987) 131–41.

    Article  PubMed  CAS  Google Scholar 

  35. Grosveld, F., van Assendelft, G. B., Greaves, D. R. & Kollias, G., Position-independent, high-level expression of the human β-globin gene in transgenic mice. Cell, 51 (1987) 975–85.

    Article  PubMed  CAS  Google Scholar 

  36. Swartz, R., Alternatives for the production of mammalian cell products: A survey. Gen. Engng News, 5 (1985) 16–21.

    Google Scholar 

  37. Colbere-Garapin, F., Horaud, F., Kourilsky, P. & Garapin, A. C., Stable HBV surface antigen expression by Vero cell clones after transfection. Develop. Biol. Standard., 59 (1985) 109–12.

    CAS  Google Scholar 

  38. Arathoon, W. R. & Birch, J. R., Large-scale cell culture in biotechnology. Science, 232 (1986) 1390–5.

    Article  PubMed  CAS  Google Scholar 

  39. Sedivy, J. M., New genetic methods for mammalian cells. Bio/Technology, 6 (1988) 1192–6.

    Article  CAS  Google Scholar 

  40. Morrison, S. L., Johnson, M. J., Herzenberg, L. A. & Oi, V. T., Chimeric human antibody molecules: Mouse antigen-binding domains with human constant region domains. Proc. Nat. Acad. Sci., USA, 81 (1984) 6851–5.

    Article  CAS  Google Scholar 

  41. Boulianne, G. L., Hozumi, N. & Shulman, M. J., Production of functional chimeric mouse/human antibody. Nature, 312 (1984) 643–6.

    Article  PubMed  CAS  Google Scholar 

  42. Boulianne, G. L., Iseman, D. E., Hozumi, N. and Shulman, M. J., Biological properties of chimeric antibodies: Interaction with complement. Molec. Biol. Med., 4 (1987) 37–49.

    PubMed  CAS  Google Scholar 

  43. Sahagan, B. G., Dorai, H., Saltzgaber-Muller, J., Toneguzzo, F., Guindon, C. A., Lilly, S. P., McDonald, K. W., Morrissey, D. V., Stone, B. A., Davis, G. L., Mcintosh, P. K. & Moore, G. P., A genetically engineered murine/human chimeric antibody retains specificity for human tumor-associated antigen. J. Immunol., 137 (1986) 1066–74.

    PubMed  CAS  Google Scholar 

  44. Sun, L. K., Curtis, P., Rakowicz-Szulczynska, E., Ghrayeb, J., Chang, N., Morrison, S. L. & Koprowski, H., Chimeric antibody with human constant regions and mouse variable regions directed against carcinoma-associated antigen 17–1A. Proc. Nat. Acad. Sci., USA, 84 (1987) 214–18.

    Article  CAS  Google Scholar 

  45. Steplewski, Z., Sun, L. K., Shearman, C. W., Ghrayeb, J., Daddona, P. & Koprowski, H., Biological activity of human-mouse IgG1, IgG2, IgG3, and IgG4 chimeric monoclonal antibodies with antitumor specificity. Proc. Nat. Acad. Sci., USA, 85 (1988) 4852–6.

    Article  CAS  Google Scholar 

  46. Nishimura, Y., Yokoyama, M., Araki, K., Veda, R., Kudo, A. & Watanabe, T., Recombinant human-mouse chimeric monoclonal antibody specific for common acute lymphocytic leukemia antigen. Cancer Res., 47 (1987) 999–1005.

    PubMed  CAS  Google Scholar 

  47. Liu, A. Y., Robinson, R. R., Hellstrom, K. E., Murray, E. D., Chang, C. P. & Hellstrom, I., Chimeric mouse-human IgG1 antibody that can mediate lysis of cancer cells. Proc. Nat. Acad. Sci., USA, 84 (1987) 3439–43.

    Article  CAS  Google Scholar 

  48. Liu, A. Y., Mack, P. W., Champion, C. I. & Robinson, R. R., Expression of mouse: human immunoglobulin heavy-chain cDNA in lymphoid cells. Gene, 54 (1987) 33–40.

    Article  PubMed  CAS  Google Scholar 

  49. Riechmann, L., Clark, M., Waldmann, H. & Winter, G., Reshaping human antibodies for therapy. Nature, 332 (1988) 323–7.

    Article  PubMed  CAS  Google Scholar 

  50. Falkner, F. G. & Zachau, H. G., Expression of mouse immunoglobulin genes in monkey cells. Nature, 298 (1982) 286–8.

    Article  PubMed  CAS  Google Scholar 

  51. Weidle, U. H., Borgya, A., Mattes, R., Lenz, H. & Buckel, P., Reconstitution of functionally active antibody directed against creatine kinase from separately expressed heavy and light chains in non-lymphoid cells. Gene, 51 (1987) 21–9.

    Article  PubMed  CAS  Google Scholar 

  52. Bodmer, M., In vitro and in vivo activities of a mouse-human chimeric B72.3 antibody. Presented at Advances in the applications of monoclonal antibodies in clinical oncology, University of London Royal Postgraduate Medical School, 25–27 May, 1988.

    Google Scholar 

  53. Vehar, G. A., Kohr, W. J., Bennett, W. F., Pennica, D., Ward, C. A., Harkins, R. N. & Collen, D., Characterization studies on human melanoma cell tissue plasminogen activator. Bio/Technology, 2 (1984) 1051–7.

    Article  Google Scholar 

  54. Ramabhadran, T. V., Reitz, B. A. & Tiemeier, D. C., Synthesis and glycosylation of the common subunit of human glycoprotein hormones in mouse cells. Proc. Nat. Acad. Sci., USA, 81 (1984) 6701–5.

    Article  CAS  Google Scholar 

  55. Stenlund, A., Lamy, D., Moreno-Lopez, J., Ahola, H., Petterson, U. & Tiollais, P., Secretion of the hepatitis B virus surface antigen from mouse cells using an extra-chromosomal eucaryotic vector. EMBOJ., 2 (1983) 669–73.

    CAS  Google Scholar 

  56. Denniston, K. J., Yoneyama, T., Hoyer, B. H. & Gerin, J. L., Expression of hepatitis B virus surface and e antigen genes cloned in bovine papilloma vectors. Gene, 32 (1984) 357–68.

    Article  PubMed  CAS  Google Scholar 

  57. Wang, Y., Stratowa, C., Schaefer-Ridder, M., Doehmer, J. & Hofschneider, P. H., Enhanced production of hepatitis B surface antigen in NIH 3T3 mouse fibroblasts by using extrachromosomally replicating bovine papillomavirus vector. Molec. Cell Biol., 3 (1983) 1032–9.

    PubMed  CAS  Google Scholar 

  58. Stratowa, C., Doehmer, J., Wang, Y. & Hofschneider, P. H., Recombinant retroviral DNA yielding high expression of hepatitis B surface antigen. EMBOJ., 1 (1982) 1573–8.

    CAS  Google Scholar 

  59. Dubois, M. F., Pourcel, C., Rousset, S., Chany, C. & Tiollais, P., Excretion of hepatitis B surface antigen particles from mouse cells transformed with cloned viral DNA. Proc. Nat. Acad. Sci., USA, 77 (1980) 4549–53.

    Article  CAS  Google Scholar 

  60. Gough, N. M. & Murray, K., Expression of the hepatitis B virus surface, core and e antigen genes by stable rat and mouse cell lines. J. Molec, Biol., 162 (1982) 43–67.

    Article  CAS  Google Scholar 

  61. Malpiece, Y., Michael, M. L., Carloni, G., Revel, M., Tiollais, P. & Weissenbach, J., The gene S promoter of hepatitis B virus confers constitutive gene expression. Nucl. Acids Res., 11 (1983) 4645–54.

    Article  PubMed  CAS  Google Scholar 

  62. Carloni, G., Malpiece, Y., Michel, M. L., Le Patezour, A, Sobczak, E., Tiollais, P. & Streeck, R. E., A transformed Vero cell line stably producing the hepatitis B virus surface antigen. Gene, 31 (1984) 49–57.

    Article  PubMed  CAS  Google Scholar 

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  1. Medical Research Council, Collaborative Centre, Mill Hill, London, UK

    Mary M. Bendig

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  1. Glaxo Group Research Limited, Greenford Road, Greenford, Middlesex, UB6 OHE, UK

    T. J. R. Harris (Director of Biotechnology) (Director of Biotechnology)

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© 1990 Elsevier Science Publishers Ltd

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Bendig, M.M. (1990). Strategies for Expressing Cloned Genes in Mammalian Cells. In: Harris, T.J.R. (eds) Protein Production by Biotechnology. Springer, Boston, MA. https://doi.org/10.1007/978-1-4613-1565-0_6

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  • DOI: https://doi.org/10.1007/978-1-4613-1565-0_6

  • Publisher Name: Springer, Boston, MA

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  • Online ISBN: 978-1-4613-1565-0

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