Abstract
Genetic drug-resistance markers were transferred via purified metaphase chromosomes from mouse L cells into the human fibrosarcoma line HT1080 and HeLa S3 cells. Interspecific chromosome-mediated transfer of hypoxanthine-guanine phosphoribosyl transferase (HGPRT; EC 2.4.2.8) from mouse L cells into HGPRT− HT1080 cells occurred at a frequency of approximately 1×10−7. The presence of the mouse allele for HGPRT in transferent isolates was confirmed by isoelectric focusing. Transfer of ouabain resistance from mouse L cells to HT1080 and HeLa S3 cells occurred at an average frequency of approximately 4×10−7. Expression of the mouse trait in transferent isolates was confirmed by their ability to withstand doses of ouabain which would be lethal to spontaneous ouabain-resistant mutants of the human cells but not to mouse L cells. Ouabain-resistant transferents of human cells showed 104- to >105-fold enhanced drug resistance, characteristic of either wild-type or mutant alleles, respectively, from ouabain-resistant donor L cells. Unstable expression of the transferred phenotypes in the absence of selection was seen in some isolates, but expression was lost at slow rates.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Baker, R. M., Brunette, D. M., Mankovitz, R., Thompson, L. H., Whitmore, G. F., Siminovitch, L., and Till, J. E. (1974). Ouabain-resistant mutants of mouse and hamster cells in culture. Cell 19.
Baker, R. M., Van Voorhis, W. C., and Spencer, L. A. (1979). HeLa cell variants that differ in sensitivity to monofunctional alkylating agents, with independence of cytotoxic and mutagenic responses. Proc. Natl. Acad. Sci. USA 765249.
Baker, R. M. (1976). Genetic and cellular properties of ouabain-resistant mutants. In Cook, J. S. (ed.), Biogenesis and Turnover of Membrane Macromolecules Raven Press, New York, pp. 93–103.
Capecchi, M. (1980). High efficiency transformation by direct microinjection of DNA into cultured mammalian cells. Cell 22479.
Chapman, V. M., Kratzer, P. G., Siracusa, L. D., Quarantillo, B. A., Evans, R., and Liskay, R. M. (1982). Evidence for DNA modification in the maintenance of X-chromosome inactivation of adult mouse tissues. Proc. Natl. Acad. Sci. USA 795357.
Fenwick, R. G., and Caskey, C. T. (1975). Mutant Chinese hamster cells with a thermosensitive hypoxanthine-guanine phosphoribosyltransferase. Cell 5115.
Fournier, R. E. K., and Ruddle, F. H. (1977). Microcell-mediated transfer of murine chromosomes into mouse, Chinese hamster and human somatic cells. Proc. Natl. Acad. Sci. USA 74319.
Hudson, L. D., Erbe, R. W., and Jacoby, L. B. (1980). Expression of the human argininosuccinate synthetase gene in hamster transferents. Proc. Natl. Acad. Sci. USA 774234.
Johnson, G. G., Eisenberg, L. R., and Migeon, B. R. (1979). Human and mouse hypoxanthine-guanine phosphoribosyltransferase: Dimers and tetramers. Science 203174.
Klobutcher, L. A., and Ruddle, F. H. (1979). Phenotype stabilization and integration of transferred material in chromosome-mediated gene transfer. Nature 280657.
Klobutcher, L. A., Miller, C. L., and Ruddle, F. H. (1980). Chromosome-mediated gene transfer results in two classes of unstable transformants. Proc. Natl. Acad. Sci. USA 773610.
Kozak, C. A., Fournier, R. E. K., Leinwand, L. A., and Ruddle, F. H. (1979). Assignment of the gene governing cellular ouabain-resistance to Mus musculus chromosome 3 using human/mouse microcell hybrids. Biochem. Genet. 1723.
Kucherlapati, R. S., Baker, R. M., and Ruddle, F. H. (1975). Ouabain as a selective agent in the isolation of somatic cell hybrids. Cytogenet. Cell. Genet. 14362.
Lewis, W. H. (1983). Chromosome-mediated gene transfer with the Chinese hamster ovary cell line. Exp. Cell Res. 143309.
Lewis, W. H., and Srinivasan, P. (1983). Chromosome-mediated gene transfer of hydroxyurea-resistance and amplification of ribonucleotide reductase activity. Mol. Cell Biol. 31053.
Lewis, W. H., Srinivasan, P. R., Stokoe, N., and Siminovitch, L. (1980). Parameters governing the transfer of the genes for thymidine kinase and dihydrofolate reductase into mouse cells using metaphase chromosomes or DNA. Somat. Cell Genet. 6333.
Lugo, T. G., and Baker, R. M. (1983). Chromosome-mediated gene transfer of HPRT and APRT in an intraspecific human cell system. Somat. Cell Genet. 9175.
Meunier-Rotival, M., Soriano, P., Cuny, G., Strauss, F., and Bernardi, G. (1982). Sequence organization and genomic distribution of the major family of interspersed repeats of mouse DNA. Proc. Natl. Acad. Sci. USA 79355.
Miller, C. L., and Ruddle, F. H. (1978). Co-transfer of human X-linked markers into murine somatic cells via isolated metaphase chromosomes, Proc. Natl. Acad. Sci. USA 753346.
Muneer, R. S., and Gray, P. N. (1979). Alteration of human breast tumor cell membrane functions by chromosome-mediated gene transfer. J. Supramol. Struct. 12355.
Peterson, J. L., and McBride, O. W. (1980). Cotransfer of linked eukaryotic genes and efficient transfer of hypoxanthine phosphoribosyltransferase by DNA-mediated gene transfer. Proc. Natl. Acad. Sci. USA 771583.
Robins, D. M., Ripley, S., Henderson, A.S., and Axel, R. (1981). Transforming DNA integrates into the host chromosome. Cell. 2329.
Rasheed, S., Nelson-Rees, W. A., Toth, E. M., Arnstein, P., and Gardner, M. B. (1974). Characterization of a newly-derived human sarcoma cell line. Cancer 331027.
Shih, C., Shilo, B. Z., Goldfarb, M. P., Dannenberg, A., and Weinberg, R. A. (1979). Passage of phenotypes of chemically transformed cells via transfection of DNA and chromatin. Proc. Natl. Acad. Sci. USA 765714.
Shatkin, A. J. (1969). Colorimetric reactions for RNA, DNA and protein determinations. In Habel, K., and Salzman, N. P. (eds.), Fundamental Techniques in Virology Academic Press, New York, pp. 231–237.
Srinivasan, P., and Lewis, W. H. (1980). Transfer of the dihydrofolate reductase gene into mammalian cells using metaphase chromosomes or purified DNA. In Baserga, R., Croce, C., and Rovera, G. (eds.), Introduction of Macromolecules into Viable Mammalian Cells Alan R. Liss, New York.
Thompson, L. H., and Baker, R. M. (1973). Isolation of mutants in cultured mammalian cells. In Prescott, D. M. (ed.), Methods in Cell Biology, Vol. 6 Academic Press, New York, pp. 209–281.
Wigler, M., Pellicer, A., Silverstein, S., Axel, R., Urlaub, G., and Chasin, L. (1979). DNA-mediated transfer of the adenine phosphoribosyltransferase locus into mammalian cells. Proc. Natl. Acad. Sci. USA 761373.
Willecke, K., and Ruddle, F. H. (1975). Transfer of the human gene for hypoxanthine phosphoribosyltransferase via isolated human metaphase chromosomes into mouse L-cells. Proc. Natl. Acad. Sci. USA 721792.
Author information
Authors and Affiliations
Additional information
This work was supported by National Institutes of Health Grant GM30383/21665 to RMB, Core Grants CA14051 to S. E. Luria and CA24538 to E. Mihich, and institutional predoctoral Training Grant GM07287.
Rights and permissions
About this article
Cite this article
Lugo, T.G., Baker, R.M. Chromosome-mediated transfer of murine alleles for hypoxanthine-guanine phosphoribosyl transferase (HGPRT) and ouabain resistance into human cell lines. Biochem Genet 23, 1–15 (1985). https://doi.org/10.1007/BF00499108
Received:
Revised:
Issue Date:
DOI: https://doi.org/10.1007/BF00499108