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Transformation and Expression of TK Sequences

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Somatic Cell Genetics

Part of the book series: NATO Advanced Study Institutes Series ((NSSA,volume 50))

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Abstract

Cells lacking thymidine kinase (Ltk-)can be converted to the tk+ phenotype following exposure to UV-irradiated herpes simplex virus (HSV) and selection in HAT medium (1,2,3,). These experiments suggested to us that isolation of the virus sequences encoding tk was feasible. Accordingly, virus DNA cleaved with a variety of restriction endonucleases was assayed for its ability to convert tk- cells to a HAT resistant phenotype using the calcium phosphate precipitation technique (4). Digestion of HSV DNA with Bam H I resulted in the appearance of numerous colonies following application of the DNA and selection in HAT (5). Subsequent analysis of Bam H I cleaved, size-fractionated DNA identified a unique 3.5 kb fragment that contained the information required to convert tk- cells to the tk+ phenotype. Colonies that arise following application of DNA and HAT selection are said to be transformed. Transformation results from integration of the virus and carrier DNA sequences into cellular DNA (6,7,8,). The cells’ transcriptional and translational apparatus recognize the virus tk sequences to form a functional virus specified tk.

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References

  1. Munyon, W., Kraiselburd, E., Davis, D. and Mann, R. (1971) Transfer of thymidine kinase to thymidine kinaseless L-cells by infection with ultraviolet-irradiated herpes simplex virus J. Virol., 7, 813

    PubMed  CAS  Google Scholar 

  2. Davidson, R.L., Adelstein, S.J. and Oxman, M. (1973) Herpes simplex virus as a source of thymidine kinase for thymidine kinase deficient mouse cells: Supression and reactivation of the viral enzyme. Proc. Natl. Acad. Sci. U.S.A., 70, 1912

    Article  PubMed  CAS  Google Scholar 

  3. Munyon, W., Buchsbaum, R., Paoletti, E., Mann, J., Kraiselburd, E. and Davis, D. (1972) Electrophoresis of thymidine kinase activity synthesized by cells transformed by herpes simplex virus. Virology, 49, 683

    Article  PubMed  CAS  Google Scholar 

  4. Graham, F.L. and van der Eb, A.I. (1973) A new technique for the assay of infectivity of human adenovirus 5 DNA. Virology, 52, 456

    Article  PubMed  CAS  Google Scholar 

  5. Wigier, M., Silverstein, S., Lee, L.S., Pellicer, A., Cheng, Y.C. and Axel, R. (1977) Transfer of purified herpes virus thymidine kinase gene to cultured mouse cells. Cell, 11, 223

    Article  Google Scholar 

  6. Pellicer, A., Wigier, M., Axel, R. and Silverstein, S. (1978) The transfer and stable integration of the HSV thymidine kinase gene into mouse cells. Cell, 14, 133

    Article  PubMed  CAS  Google Scholar 

  7. Robins, D.M., Ripley, S., Henderson, A.S. and Axel, R. (1981) Transforming DNA integrates into the host chromosome. Cell, 23, 29

    Article  PubMed  CAS  Google Scholar 

  8. Perucho, M., Hanahan, D. and Wigier, M. (1980) Genetic and physical linkage of exogenous sequences in transformed cells. Cell, 21, 309

    Article  Google Scholar 

  9. McKnight, S.L. (1980) The nucleotide sequence and transcript map of the herpes simplex virus thymidine kinase gene. Nucl. Acids Res., 8, 5949

    Article  PubMed  CAS  Google Scholar 

  10. Wagner, M.J., Sharp, J.A. and Summers, W.C. (1981) Nucleotide sequence of the thymidine kinase gene of herpes simplex virus type-1. Proc. Natl. Acad. Sci. U.S.A., 78, 1441

    Article  PubMed  CAS  Google Scholar 

  11. Smiley, J.R., Wagner, M.J., Summers, W.P. and Summers, W.C. (1980) Genetic and physical evidence for the polarity of the thymidine kinase gene of herpes simplex virus. Virology, 102, 83

    Article  PubMed  CAS  Google Scholar 

  12. McKnight, S.L. and Gavis, E.R. (1980) Expression of the herpes thymidine kinase gene in Xenopus laevis oocytes: An assay for the study of deletion mutants. Nucl. Acids Res., 8, 5931

    Article  PubMed  CAS  Google Scholar 

  13. McKnight, S.L., Gavis, E.R., Kingsbury, R. and Axel, R. Analysis of the HSV thymidine kinase gene: Identification of an upstream control region, (submitted for publication)

    Google Scholar 

  14. Lin, S.S. and Munyon, W. (1974) Expression of the viral thymidine kinase gene in herpes simplex virus transformed L-cells. J. Virol., 14, 1199

    PubMed  CAS  Google Scholar 

  15. Garfinkle, B. and McAuslan, B.R. (1974) Regulation of herpes simplex virus induced thymidine kinase. Biochem. Biophys. Res. Commun., 58, 822

    CAS  Google Scholar 

  16. Leiden, J.M., Buttyan, R. and Spear, P.G. (1976) Herpes simplex virus gene expression in transformed cells 1: Regulation of the viral thymidine kinase gene in transformed L cells by products of superinfecting virus. J. Virol., 20, 413

    PubMed  CAS  Google Scholar 

  17. Kit, S., Dubbs, D.R. and Schaffer, P.A. (1978) Thymidine kinase activity of biochemically transformed mouse cells after superinfection by thymidine kinase negative temperature sensitive herpes simplex virus mutants. Virology, 85, 456

    Article  PubMed  CAS  Google Scholar 

  18. Preston, C.M. (1979) Control of herpes simplex virus type 1 mRNA synthesis in cells infected with wild-type virus or the temperature sensitive mutant tsK. J. Virol., 29, 275

    PubMed  CAS  Google Scholar 

  19. Colbere-Garapin, F., Chousterman, S., Horodniceanu, F., Kourilsky, P. and Garapin, A. (1979) Cloning of the active thymidine kinase gene of herpes simplex virus type 1 in Escherichia coli K-12. Proc. Natl. Acad. Sci. U.S.A., 76, 3755

    Google Scholar 

  20. Southern, E.M. (1975) Detection of specific sequences among DNA fragments separated by gel electrophoresis. J. Mol. Biol., 98, 503

    Article  PubMed  CAS  Google Scholar 

  21. Efstradiatis, A., Posakony, J.W., Maniatis, T., Lawn, R.M., O’Connell, C., Spritz, R.A., DeRiel, J., Forget, B.G., Weissman, S.A., Slightom, J.L., Blechl, A.E., Smithies, O., Baralle, F.E., Shaulders, C.C. and Proudfoot, N.J. (1980) The structure and evolution of the human β-globin gene family. Cell, 21, 653

    Article  Google Scholar 

  22. Smiley, J. (1980) Construction in vitro and rescue of a thymidine kinase deficient deletion mutant of herpes simplex virus. Nature, 285, 333

    Article  PubMed  CAS  Google Scholar 

  23. Sweet, R., Jackson, J., Lowy, I., Ostrander, M., Pellicer, A., Roberts, J., Robins, D., Sim, G.-K., Wold, B., Axel, R. and Silverstein, S. (1981) The expression, arrangement and rearrangement of genes in DNA-transformed cells, in Genes, Chromosomes and Neoplasia, F.E. Arrighi, P.N. Rao, and E. Stubblefield, ed. Raven Press, N.Y.

    Google Scholar 

  24. McGhee, J. and Ginder, M. (1979) Specific DNA methylation sites in the vicinity of the chicken β-globin genes. Nature, 280, 419

    Article  PubMed  CAS  Google Scholar 

  25. Weintraub, H. and Groudine, M. (198.1) α-Globin gene switching during the development of chicken embryos: Expression and chromosome structure. Cell, in press

    Google Scholar 

  26. Taylor, S.M. and Jones, P.A. (1979) Multiple new phenotypes induced in 10T1/2 and 3T3 cells treated with 5-azacytidine. Cell, 17, 771

    Article  PubMed  CAS  Google Scholar 

  27. Jones, P.A. and Taylor, S.M. (1980) Cellular differentiation, cytidine analogs and DNA methylation. Cell, 20, 85

    Article  PubMed  CAS  Google Scholar 

  28. Rothstein, S.J., Jorgensen, R.A. Postle. K. and Reznikoff, W.S. (1980) The inverted repeats of Tn5 are functionally different. Cell, 19, 795

    Article  PubMed  CAS  Google Scholar 

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Authors and Affiliations

  1. College of Physicians and Surgeons, Columbia University, New York, N.Y., 10032, USA

    Adele El Kareh, Michael Ostrander & Saul Silverstein

Authors
  1. Adele El Kareh
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  2. Michael Ostrander
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  3. Saul Silverstein
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Editor information

Editors and Affiliations

  1. Howard Hughes Medical Institute, Baylor College of Medicine, Houston, Texas, USA

    C. Thomas Caskey  & D. Christopher Robbins  & 

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© 1982 Plenum Press, New York

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Kareh, A.E., Ostrander, M., Silverstein, S. (1982). Transformation and Expression of TK Sequences. In: Caskey, C.T., Robbins, D.C. (eds) Somatic Cell Genetics. NATO Advanced Study Institutes Series, vol 50. Springer, Boston, MA. https://doi.org/10.1007/978-1-4684-4256-4_8

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  • DOI: https://doi.org/10.1007/978-1-4684-4256-4_8

  • Publisher Name: Springer, Boston, MA

  • Print ISBN: 978-1-4684-4258-8

  • Online ISBN: 978-1-4684-4256-4

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