Abstract
Human DNA purified from HeLa cells and from three strains of skin fibroblasts was precipitated with calcium phosphate and added to mouse cells that were deficient in adenine phosphoribosyltransferase (APRT) and hypoxanthine phosphoribosyltransferase (HPRT). Selection for cells possessing either of the phosphoribosyltransferases was imposed by blocking de novo synthesis of purine nucleotides with azaserine in a medium supplemented with adenine and hypoxanthine. The frequency of colony formation after selection was 1.7×10−7−3.3×10−6. Excepting some azaserine-resistant colonies that appeared only in the first experiment and infrequent revertants expressing mouse APRT, all characterized clones expressed the human forms of APRT or HPRT according to the criteria of specific immunoprecipitation and electrophoretic mobility. The frequency of transfer of the human APRT gene was much greater than that of HPRT. Transfer efficiency was not significantly reduced when HeLa DNA was sheared to 6.5–13.5 kb size or when the donor DNA was isolated from a transferent that expressed human APRT.
Similar content being viewed by others
Literature cited
Szybalska, E. H., and Szybalski, W. (1962).Proc. Natl. Acad. Sci. U.S.A. 48:2026–2034.
Graham, F. L., Abrahams, P. J., Mulder, C., Heyneker, H. L., Warnaar, S. O., de Vries, F. A. J., Fiers, W., and van der Eb, A. J. (1975).Cold Spring Harbor Symp. Quant, Biol. 39:637–650.
McCutcheon, J. H., and Pagano, J. S. (1968).J. Natl. Cancer Inst. 41:351–357.
Wigler, M., Silverstein, S., Lee, L. S., Pellicer, A., Cheng, Y. C., and Axel, R. (1977).Cell 11:223–232.
Wigler, M., Pellicer, A., Silverstein, S., and Axel, R. (1978).Cell 14:725–731.
Wigler, M., Pellicer, A., Silverstein, S., Axel, R., Urlaub, G., and Chasin, L. (1979).Proc. Natl. Acad. Sci. U.S.A. 76:1373–1376.
Willecke, K., Klomfass, M., Mierau, R., and Dohmer, J. (1979).Mol. Gen. Genet. 170:179–185.
Kahan, B., and DeMars, R. (1975).Proc. Natl. Acad. Sci. U.S.A. 72:1510–1514.
Kit, S., Dubbs, D., Piekarski, L., and Hsu, T. (1963).Exp. Cell Res. 31:297–312.
Bang, F. B. (1977).Bull. Hist. Med. 51:516–537.
DeMars, R., and Held, K. R. (1972).Humangenetik 16:87–110.
Ham, R. G. (1963).Exp. Cell Res. 29:515–526.
Rappaport, H., and DeMars, R. (1973).Genetics 75:335–345.
Oyama, V. I., and Eagle, H. (1956).Proc. Soc. Exp. Biol. Med. 91:305–307.
Held, K. R., Kahan, B., and DeMars, R. (1975).Humangenetik 30:23–34.
Bakay, B., and Nyhan, W. L. (1971).Biochem. Genet. 5:81–90.
Shinnick, T. M., Lund, E., Smithies, O., and Blattner, F. R. (1975).Nucleic Acids Res. 2:1911–1929.
Degnen, G. E., Miller, I. L., Adelberg, E. A., and Eisenstadt, J. M. (1976).Proc. Natl. Acad. Sci. U.S.A. 73:2838–2842.
Wullems, G. J., van der Horst, J., and Bootsma, D. (1976).Somat. Cell Genet. 2:155–164.
Davies, P. J., and Willecke, K. (1977).Mol. Gen. Genet. 154:191–197.
Willecke, K., and Ruddle, F. H. (1975).Proc. Natl. Acad. Sci. U.S.A. 72:1792–1796.
Wullems, G. J., van der Horst, J., and Bootsma, D. (1976).Somat. Cell Genet. 2:359–371.
Burch, J. W., and McBride, O. W. (1975).Proc. Natl. Acad. Sci. U.S.A. 72:1797–1801.
Lyon, M. F. (1972).Biol. Rev. 47:1–35.
Russell, L. B. (1963).Science 140:976–978.
Migeon, B. R. (1972).Nature 239:87–89.
Henderson, J. F. (1972).Am. Chem. Soc. Monog. 170 (American Chemical Society, Washington, D. C.).
Schimke, R. T., Kaufman, R. J., Alt, F. W., and Kellems, R. F. (1978).Science 202:1051–1055.
McBride, W. O., and Ozer, H. L. (1973).Proc. Natl. Acad. Sci. U.S.A. 70:1258–1262.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Lester, S.C., LeVan, S.K., Steglich, C. et al. Expression of human genes for adenine phosphoribosyltransferase and hypoxanthine-guanine phosphoribosyltransferase after genetic transformation of mouse cells with purified human DNA. Somat Cell Mol Genet 6, 241–259 (1980). https://doi.org/10.1007/BF01538799
Received:
Revised:
Issue Date:
DOI: https://doi.org/10.1007/BF01538799