Biological Activity of Simian Virus 40 DNA Fragments and T-Antigen Tested by Microinjection into Tissue Culture Cells

  • A. Graessmann
  • M. Graessmann
  • C. Mueller
Part of the NATO Advanced Study Institutes Series book series (NSSA, volume 31)


The biological activity of macromolecules (DNA, RNA, protein) can be directly tested by microinjection into tissue culture cells (1, 2, 3). The transfer of the test material is performed with microglass capillaries under a phase contrast microscope at a 400 fold magnification. Technical details of the method have been described elsewhere (4, see also article by Celis, Kaltoft and Bravo). In this article, mapping of early Simian Virus 40 (SV40) specific functions and studies on the activity of purified SV40 T-antigen and T-antigen related proteins are used to demonstrate some aspects of the microinjection technique.


Simian Virus Tissue Culture Cell Helper Function Actin Cable Mouse Kidney Cell 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. 1).
    Graessmann, A. and Graessmann, M. (1971). Ueber die Bildung von Melanin in Muskelzellen nach der direkten Uebertragung von RNA aus Harding-Passey-Melanomzellen. Hoppe-Seyler’s Zeit. Physiol. Chem., 352, 527.CrossRefGoogle Scholar
  2. 2).
    Graessmann, M. and Graessmann, A. (1976). Early simian virus 40 specific RNA contains information for tumor antigen formation and chromatin replication. Proc. Natl. Acad. Sci. USA., 73, 366.PubMedCrossRefGoogle Scholar
  3. 3).
    Tjian, R., Fey, G. and Graessmann, A. (1978). Biological activity of purified simian virus 40 T-antigen proteins. Proc. Natl. Acad. Sci. U.S.A., 75, 1279.PubMedCrossRefGoogle Scholar
  4. 4).
    Graessmann, A., Graessmann, M. and Mueller, C. (1979). Microinjection of SV40 nucleic acids and SV40 T-antigen. Methods in Enzymology. (eds., L. Grossman, and K. Moldave). Academic Press, New York, Vol. 65. (in press).Google Scholar
  5. 5).
    Tooze, J. (1973). The molecular biology of tumor viruses. Cold Spring Harbor Laboratory, Cold Spring Harbor, New York.Google Scholar
  6. 6).
    Crawford, L.V., Cole, C.N., Smith, A.E., Tegtmeyer, P., Rundell, K. and Berg, P. (1978). Organization and expression of early genes of simian virus 40. Proc. Natl. Acad. Sci. U.S.A., 75, 117.PubMedCrossRefGoogle Scholar
  7. 7).
    Prives, C., Gilboa, E., Revel, M. and Winoccour, E. (1977). Cell-free translation of simian virus 40 early messenger RNA coding for viral T-antigen. Proc. Natl. Acad. Sci. U.S.A., 74, 457.PubMedCrossRefGoogle Scholar
  8. 8).
    Weil, R., Salomon, C., May, E. and May, P. (1974). A simplifying concept in tumor virology: virus-specific “pleiotropic” effectors, Cold Spring Harbor Symp. Quant. Biol., 39, 381.CrossRefGoogle Scholar
  9. 9).
    Tegtmeyer, P. (1972). Simian virus 40 deoxyribonucleic acid synthesis: the viral replicon. J. Virol., 10, 591.PubMedGoogle Scholar
  10. 10).
    Risser, R. and Pollack, R. (1974). A nonselective analysis of SV40 transformation of mouse 3T3 cells. Virology, 59, 477.PubMedCrossRefGoogle Scholar
  11. 11).
    Osborn, M. and Weber, K. (1975). Simian virus 40 gene A function and maintenance of transformation. J. Virol., 15, 636.PubMedGoogle Scholar
  12. 12).
    Tegtmeyer, P. (1975). Function of simian virus 40 gene A in transforming infection. J. Virol., 8, 613Google Scholar
  13. 13).
    Graessmann, A., Graessmann, M., Hoffmann, H., Niebel, J., Brandner, G. and Mueller, N. (1974). Inhibition by interferon of SV40 tumor antigen formation in cells injected with SV40 cRNA transcribed in vitro. FEBS Lett., 39, 249.PubMedCrossRefGoogle Scholar
  14. 14).
    Mueller, C., Graessmann, A. and Graessmann, M. (1978). Mapping of early SV40 specific functions by microinjection of different early viral DNA fragments. Cell, 15, 579.PubMedCrossRefGoogle Scholar
  15. 15).
    Graessmann, A., Graessmann, M. and Mueller, C. (1977). Regulatory function of simian virus 40 DNA replication for late viral gene expression. Proc. Natl. Acad. Sci. U.S.A., 74, 4831.PubMedCrossRefGoogle Scholar
  16. 16).
    Graessmann, A., Graessmann, M., Guhl, E. and Mueller, C. (1978). Quantitative correlation between simian virus 40 T-antigen synthesis and late viral gene expression in permissive and non-permissive cells. J. Cell Biol., 77, R1.PubMedCrossRefGoogle Scholar
  17. 17).
    Graessmann, A., Graessmann, M. and Mueller, C. (1976). Regulatory mechanism of simian virus 40 gene expression in permissive and nonpermissive cells. J. Virol., 17, 854.PubMedGoogle Scholar
  18. 18).
    Topp, W.C., Rifkin, D., Graessmann, A., Chang, C.M. and Sleigh, M.J. (1979). The role of early SV40 gene products in the maintenance of the transformed state. Hormons and Cell Culture. (eds., G. Sato and R. Ross). Cold Spring Harbor Laboratory, Cold Spring Harbor, New York. (in press).Google Scholar
  19. 19).
    Graessmann, A., Graessmann, M., Tjian, R. and Topp, W. (1979). Small SV40 t-antigen is necessary to induce the loss of actin cable structure. (Submitted for publication).Google Scholar
  20. 20).
    Graessmann, M., Graessmann, A. and Mueller, C. (1979). SV40 DNA fragment transformed monkey cells and flat revertants synthesize large and small tumor antigens. Cold Spring Harbor Symp. Quant. Biol., 44 (in press).Google Scholar

Copyright information

© Plenum Press, New York 1980

Authors and Affiliations

  • A. Graessmann
    • 1
  • M. Graessmann
    • 1
  • C. Mueller
    • 1
  1. 1.Institut fuer Molekularbiologie und BiochemieFreien Universitaet Berlin1 Berlin 33West Germany

Personalised recommendations