Gene Transfer as a General Approach to Facilitate the Recombinant DNA Cloning of Surface Antigen Genes

  • Frank H. Ruddle


Flow cytometry and the use of monoclonal antibodies have contributed significantly to a number of somatic cell genetic methodologies involving surface antigens, such as gene mapping using somatic cell hybrids, DNA-mediated transformation of recipient cells using cloned surface antigen genes, and, most recently, surface antigen gene cloning. These topics have been previously reviewed.1 In this chapter, I shall discuss recent progress involving the recombinant DNA cloning of surface antigen genes, using the transferrin receptor as a model system.2 The use of eukaryotic cells as host systems for DNA recombinant cloning of non-membrane protein genes has been reviewed recently.3


Transferrin Receptor Recipient Cell Primary Transformants Thymidine Kinase Gene Secondary Transformants 
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  1. 1.
    Kamarck, M. E., Barbosa, J. A., Kuhn, L., Messer Peters, P., Shulman, L., and Ruddle, F. H., 1983, Somatic cell genetics and flow cytometry, Cytometry 4:99–108.PubMedCrossRefGoogle Scholar
  2. 2.
    Kuhn, L. C., McClelland, A., and Ruddle, F. H., 1984, Gene transfer expression and molecular cloning of the human transferrin receptor gene, Cell 37:95–103.PubMedCrossRefGoogle Scholar
  3. 3.
    Ruddle, F. H., Kamarck, M. E., McClelland, A., and Kuhn, L. C., 1984, DNA-mediated gene transfer in mammalian gene cloning, in: Genetic Engineering, Volume 6 (J. K. Setlow and A. Hollaender, eds.), Plenum Press, New York, pp. 319–338.Google Scholar
  4. 4.
    Schneider, C., Sutherland, R., Newman, R., and Greaves, M., 1982, Structural features of the cell surface receptor for transferrin that is recognized by the monoclonal antibody OKT9, J. Biol. Chem. 257:8516–8522.PubMedGoogle Scholar
  5. 5.
    Newman, R., Schneider, C., Sutherland, R., Vodinelich, L., and Greaves, M., 1982, The transferrin receptor, Trends Biochem. Sci. 1:397–400.CrossRefGoogle Scholar
  6. 6.
    Dautry-Varsat, A., Ciechanover, A., and Lodish, H. F., 1983, pH and the recycling of transferrin during receptor-mediated endocytosis. Proc. Natl. Acad. Sci. USA 80:2258–2262.Google Scholar
  7. 7.
    Neckers, L. M., and Cossman, J., 1983, Transferrin receptor induction in mitogen-stimulated human T lymphocytes is required for DNA synthesis and cell division and is regulated by interleukin 2, Proc. Natl Acad. Sci. USA 80:3494–3498.PubMedCrossRefGoogle Scholar
  8. 8.
    Barbosa, J. A., Kamarck, M. E., Biro, P. A., Weissman, S. M., and Ruddle, F. H., 1982, Identification of human genomic clones coding the major histocompatibility antigens HLA-A2 and HLA-B7 by DNA-mediated gene transfer, Proc. Natl. Acad. Sci. USA 79:6327–6331.PubMedCrossRefGoogle Scholar
  9. 9.
    Barbosa, J. A., Kamarck, M. E., and Ruddle, F. H., 1983, Variation in expression of human major histocompatibility genes in mouse L cells after DNA-mediated gene transfer, in: Gene Expression in Normal and Transformed Cells (J. E. Cellis and E. Bravo, eds.), Plenum Press, New York, pp. 379–401.Google Scholar
  10. 10.
    Lin, P.-F., Zhao, S.-Y., and Ruddle, F. H., 1983, Genomic cloning and preliminary characterization of the human thymidine kinase gene, Proc. Natl. Acad. Sci. USA 80:6528–6532.PubMedCrossRefGoogle Scholar
  11. 11.
    Kuhn, L. C., Barbosa, J. A., Kamarck, M. E., and Ruddle, F. H., 1983, A new approach to the cloning of cell surface protein genes: Selection by cell sorting of mouse L-cells which express HLA or 4F2 antigens after transformation with total human DNA, Mol. Biol. Med. 1:335–352.PubMedGoogle Scholar
  12. 12.
    Kavathas, P., and Herzenberg, L. A., 1983, Stable transformation of mouse L cells for human membrane T-cell differentiation antigens, HLA and β2-microglobulin: Selection by fluorescence-activated cell sorting, Proc. Natl. Acad. Sci. USA 80:524–527.PubMedCrossRefGoogle Scholar

Copyright information

© Plenum Press, New York 1985

Authors and Affiliations

  • Frank H. Ruddle
    • 1
  1. 1.Department of Biology and Human GeneticsYale UniversityNew HavenUSA

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