Skip to main content
SpringerLink
Log in

Radiation fusion hybrids for human chromosomes 3 and X generated at various irradiation doses

  • Published:
Somatic Cell and Molecular Genetics

Abstract

We have used a gamma-irradiation (2.5–25 krads) cell fusion procedure to generate human-hamster somatic cell hybrids (IHB, irradiated human fragments in B14–150 cells), retaining small fragments derived from human chromosomes 3 and X. By using Alu-element mediated PCR amplification and dot-blot hybridization with human alphoid or total human DNA as probes, 86 positive hybrids were identified and selected for further analysis. Nonisotopic fluorescence in situ hybridization (FISH) with human DNA in a set of eight hybrids demonstrated the presence of from one to eight human fragments per cell independent of irradiation dose. In contrast, a significant dose-dependent variation of fragment sizes was shown in the analysis of the 86 hybrids with markers previously mapped to 3p (seven markers) and to Xq (21 markers). Using the Xq27–28 region as a model, 40% of the hybrids generated at 5 krads or less were found to have retained fragments in the range of 3–30 Mb, 10% retained the whole chromosome arm, and the remaining 50% retained fragments of less than 2–3 Mb. The proportion of fragments of 3 Mb or larger decreased rapidly at higher irradiation doses and was very low (less than 6%) in hybrids generated at 25 krads. Upon further characterization, the 86 hybrids analyzed here will provide a mapping panel for the entire chromosomes 3 and X with an estimated resolution in the range of 1–2 Mb on average, a size range amenable to PFGE and YAC contig mapping.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

Literature cited

  1. Röhme, D., Fox, H., Hermann, B., Frischauf, A.-M., Edström, J.-E., Mains, P., Silver, L., and Lehrach, H. (1984).Cell 36783–788.

    PubMed  Google Scholar 

  2. Ludecke, H.-J., Senger, G., Claussen, U., and Horsthemke, B. (1989).Nature 338348–350.

    PubMed  Google Scholar 

  3. Kunkel, L.M., Monaco, A.P., Middlesworth, W., Ochs, H.D., and Latt, S. (1985).Proc. Natl. Acad. Sci. U.S.A. 824778–4782.

    PubMed  Google Scholar 

  4. Gusella, J.F., Keys, C., Varsanyi-Brenner, A., Kao, F.-T., Jones, C., Puck, T.T., and Housman, D. (1980).Proc. Natl. Acad. Sci. U.S.A. 772829–2833.

    PubMed  Google Scholar 

  5. Smith, D.I., Mangrulker, R., Geist, R., Gilbert, J., Kinsman, K., Drabkin, H., and Golembieski, W. (1989).Genomics 4453–459.

    PubMed  Google Scholar 

  6. Nelson, D.L., Ledbetter, S.A., Corbo, L., Victoria, M.F., Ramirez-Solis, R., Webster, T.D., Ledbetter, D.H., and Caskey, C.T. (1989).Proc. Natl. Acad. Sci. U.S.A. 866686–6690.

    PubMed  Google Scholar 

  7. Ledbetter, S., Nelson, D., Warren, S., and Ledbetter, D. (1990).Genomics 6475–481.

    PubMed  Google Scholar 

  8. Klobutcher, L., and Ruddle, F. (1979).Nature 280657–660.

    PubMed  Google Scholar 

  9. Goss, S.J., and Harris, H. (1975).Nature 255680–684.

    PubMed  Google Scholar 

  10. McBride, O.W., and Ozer, H.L. (1973).Proc. Natl. Acad. Sci. U.S.A. 701258–1262.

    PubMed  Google Scholar 

  11. Miller, C.L., and Ruddle, F.H. (1978).Proc. Natl. Acad. Sci. U.S.A. 753987–3990.

    Google Scholar 

  12. Bickmore, W., Christie, S., van Heyningen, V., Hastie, N.D., and Porteous, D.J. (1988).Nucleic Acids Res. 1651–61.

    PubMed  Google Scholar 

  13. Xu, W., Gorman, P., Rider, S., Hedge, P., Moore, G., Pritchard, C., Sheer, D., and Solomon, E. (1988).Proc. Natl. Acad. Sci. U.S.A. 858363–8367.

    Google Scholar 

  14. Glaser, T., Rose, E., Morse, H., Housman, D., and Jones, C. (1990).Genomics 648–64.

    PubMed  Google Scholar 

  15. Zoghbi, H.Y., McCall, A.E., and LeBorgne-Demarquoy, F. (1991).Genomics 9713–720.

    PubMed  Google Scholar 

  16. Goodfellow, P.J., Povey, S., Nevanlinna, H.A., and Godfellow, P.N. (1990).Somat. Cell Mol. Genet. 16163–171.

    PubMed  Google Scholar 

  17. Cox, D.R., Burmeister, M., Price, E.R., Kim, S., and Meyers, R.M. (1990).Science 250246–250.

    Google Scholar 

  18. Burmeister, M., Kim, S., Price, E.R., de Lange, T., Tantravahi, U., Meyers, R.M., and Cox, D.R. (1991).Genomics 919–30.

    PubMed  Google Scholar 

  19. Yerganian, G., and Leonard, M. (1961).Science 1331600–1601.

    PubMed  Google Scholar 

  20. Sidén, T.S., Höglund, M., and Röhme, D. (1990).Somat. Cell Mol. Genet. 16425–435.

    PubMed  Google Scholar 

  21. Feinberg, A.P., and Vogelstein, B. (1983).Anal. Biochem. 1326–13.

    PubMed  Google Scholar 

  22. Gray, K.M., White, J.W., Costoni, C., Gillespie, D., Schroeder, W.T., Calabretta, B., and Sounders, G. (1985).Nucleic Acids Res. 13521–535.

    PubMed  Google Scholar 

  23. Suthers, G.K., Hyland, V.J., Callen, D.F., Oberle, I., Rocchi, M., Thomas, N.S., Morris, C.P., Schwartz, C.E., Schmidt, M., Ropers, H.H., Baker, E., Oostra, B.A., Dahl, N., Wilson, P.J., Hopwood, J.J., and Sutherland, G.R. (1989).Am. J. Hum. Genet. 47187–195.

    Google Scholar 

  24. Suthers, G.K., Mulley, J.C., Voelckel, M.A., Dahl, N., Väisänen, M.L., Steinbach, P., Glass, I.A., Schwartz, C.E., van Oost, B.A., Thibodeau, S.N., Haites, N.E., Oostra, B.A., Giné, R., Carballo, M., Morris, C.P., Hopwood, J.J., and Sutherland, G.R. (1991).Am. J. Hum. Genet. 48460–467.

    PubMed  Google Scholar 

  25. Benham, F., Hart, K., Crolla, J., Bobrow, M., Francavilla, M., and Goodfellow, P.N. (1989).Genomics 4509–517.

    PubMed  Google Scholar 

  26. Cox, D.R., Pritchard, C.A., Uglum, E., Casher, D., Kobori, J., and Meyers, R.M. (1989).Genomics 4397–407.

    PubMed  Google Scholar 

  27. Cirullo, R.E., Dana, S., and Wasmuth, J.J. (1983).Mol. Cell. Biol. 3892–902.

    PubMed  Google Scholar 

  28. Brooks-Wilson, A.R., Goodfellow, P.N., Povey, S., Nevanlinna, H.A., de Jong, P.J., and Goodfellow, P.J. (1990).Genomics 7614–620.

    PubMed  Google Scholar 

  29. Bonner, T., O'Brian, J., Nash, W., Rapp, U., Morton, C., and Leder, P. (1984).Science 22371–74.

    PubMed  Google Scholar 

  30. Albertson, D.G., Sherrington, P.D., and Rabbits, P.H. (1989).Hum. Genet. 83127–132.

    PubMed  Google Scholar 

  31. Barker, D., Schafter, M., and White, R. (1984).Cell 36131–138.

    PubMed  Google Scholar 

  32. Brauch, H., Kalman, T., Kotler, F., Gazdar, A.F., Pettengill, O.S., Johnson, B., Graziano, S., Winton, T., Buys, C.H.C.M., Sorenson, G.D., Poiesz, B.J., Minna, J.D., and Zbar, B. (1990).Genes Chrom. Cancer 1240–246.

    PubMed  Google Scholar 

  33. Hofker, M.H., Skraastad, M.I., Bergen, A.A.B., Wapenaar, M.C., Bakker, E., Millington-Ward, A., van Ommen, G.J.B., and Pearson, P.L. (1986).Am. J. Hum. Genet. 39438–451.

    PubMed  Google Scholar 

  34. Bruns, G., Aldridge, J., Kunkel, L., Trantravahi, U., Lalande, M., Dryja, T., and Latt, S.A. (1984).Cytogenet. Cell Genet. 37428–429.

    Google Scholar 

  35. Hofker, M.H., Bergen, A.A.B., Skraastad, M.I., Carpenter, N.J., Veenema, H., Connor, J.M., Bakker, E., van Ommen, G.J.B., and Pearson, P.L. (1987).Am. J. Hum. Genet. 40312–328.

    PubMed  Google Scholar 

  36. Schmeckpeper, B.J., Davis, J., Willard, H.F., and Smith, K.D. (1985).Nucleic Acids Res. 135724.

    PubMed  Google Scholar 

  37. Page, D., De Martinville, B., Barker, D., Wyman, A., White, R., Franke, U., and Botstein, D. (1982).Proc. Natl. Acad. Sci. U.S.A. 795352–5356.

    PubMed  Google Scholar 

  38. Aldridge, J., Kunkel, L., Bruns, G., Tantravahi, U., Lalande, M., Brewster, T., Moreau, E., Wilson, M., Bromley, W., Roderick, T., and Latt, S.A. (1984).Am. J. Hum. Genet. 36546–564.

    PubMed  Google Scholar 

  39. Brown, C.J., Vivona, A., Parikh, S., Bedford, M., and Willard, H.F. (1989).Cytogenet. Cell Genet. 51970–971.

    Google Scholar 

  40. Drayna, D., Davies, K., Hartley, D., Mandel, J.-L., Camerino, G., Williamson, R., and White, R. (1984).Proc. Natl. Acad. Sci. U.S.A. 812836–2839.

    PubMed  Google Scholar 

  41. Reilly, D.S., Lewis, R.A., and Nussbaum, R.L. (1990).Genomics 862–70.

    PubMed  Google Scholar 

  42. Kidd, K.K., Bowcock, A.M., Schmidke, J., Track, R.K., Riciutti, F., Hutchings, G., Bale, A., Pearson, P. and Willard, W.F. (1989). Human gene mapping 10: Tenth international workshop of human gene mapping.Cytogenet. Cell Genet. 51622–947.

    PubMed  Google Scholar 

  43. Oostra, B.A., Hupkes, P.E., Perdon, L.F., van Bennekom, C.A., Bakker, E., Halley, D.J.J., Schmidt, M., Du Sart, D., Smits, A., Wieringa, B., and van Oost, B.A. (1990).Genomics 6129–132.

    PubMed  Google Scholar 

  44. Rosseau, F., Vincent, A., Oberle, I., and Mandel, J.-L. (1990).Hum. Genet. 84263–266.

    PubMed  Google Scholar 

  45. Patterson, M.N., Bell, M.V., Bloomfield, J., Flint, T., Dorkins, H., Thibodeau, S.N., Schaid, D., Bren, G., Schwartz, C.E., Wieringa, B., Ropers, H.H., Callen, D.F., Sutherland, G., Froster-Iskenius, U., Vissing, H., and Davies, K.E. (1988).Genomics 4570–578.

    Google Scholar 

  46. Gitschier, J., Drayna, D., Tuddenham, E.G.D., White, R., and Lawn, R.M. (1985).Nature 314738–740.

    PubMed  Google Scholar 

  47. Lengauer, C., Riethman, H., and Cremer, T. (1990).Hum. Genet. 861–6.

    PubMed  Google Scholar 

  48. Monaco, A.P., Lam, V.M.S., Zehetner, G., Lennon, G.G., Douglas, C., Nizetic, D., Goodfellow, P.N., and Lehrach, H. (1991).Nucleic Acids Res. 193315–3318.

    PubMed  Google Scholar 

Download references

Author information

Author notes

  1. D. Röhme

    Present address: Department of Molecular Genetics, Greenwood Genetic Center, 29646, Greenwood, South Carolina

Authors and Affiliations

  1. Department of Molecular Genetics, The Wallenberg Laboratory, University of Lund, Box 7031, S-220 07, Lund, Sweden

    T. S. Sidén

  2. Department of Tumor Immunology, The Wallenberg Laboratory, University of Lund, Box 7031, S-220 07, Lund, Sweden

    J. Kumlien & D. Röhme

  3. Department of Molecular Genetics, Greenwood Genetic Center, 29646, Greenwood, South Carolina

    C. E. Schwartz

Authors
  1. T. S. Sidén
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. J. Kumlien
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. C. E. Schwartz
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. D. Röhme
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Sidén, T.S., Kumlien, J., Schwartz, C.E. et al. Radiation fusion hybrids for human chromosomes 3 and X generated at various irradiation doses. Somat Cell Mol Genet 18, 33–44 (1992). https://doi.org/10.1007/BF01233447

Download citation

  • Received:

  • Revised:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF01233447

Keywords

Search

Navigation