Abstract
The conditions necessary to achieve high frequency transfer of the thymidine kinase and dihydrofolate reductase genes from hamster cells into mouse cells were investigated. Of the parameters examined, the length of adsorption time, input gene dosage, and treatment with dimethylsulfoxide (DMSO) were found to significantly alter the transfer frequency using either metaphase chromosomes or purified DNA as the transfer vehicle. With the mouse cell line as a recipient, the optimal adsorption period for DNA or chromosomes from MtxRIII cells was found to vary from 8 to 16 h in those experiments where the recipient cells were subsequently treated with DMSO. Without DMSO, similar frequencies could be obtained by extending the period of adsorption. Increasing the dosage of DNA or chromosomes resulted in an almost linear increase in the number of transformants. The optimal conditions for transfer did not significantly differ for the two genes studied. On the average, the optimal conditions yielded 1.5×103 transformants per 107 recipient cells with chromosomes; with DNA an average of only 60 transformants were observed. In general, DNA transformants grown in the absence of methotrexate were unstable; whereas, under the same conditions about 20% of the transformants from the chromosome experiments were stable.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
Literature cited
McBride, O. W., and Ozer, H. L. (1973).Proc. Natl. Acad. Sci. U.S.A. 70:1258–1262.
Miller, C. L., and Ruddle, F. H. (1978).Proc. Natl. Acad. Sci. U.S.A. 75:3346–3350.
Willecke, K., Lange, R., Kruger, A., and Reber, T. (1976).Proc. Natl. Acad. Sci. U.S.A. 73:1274–1278.
Wullems, G. J., Van Der Horst, J., and Bootsma, D. (1977).Somat. Cell Genet. 3:281–293.
McBride, O. W., Burch, J. W., and Ruddle, F. H. (1978).Proc. Natl. Acad. Sci. 75:914–918.
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. 76:1373–1376.
Spandidos, D., and Siminovitch, L. (1977).Proc. Natl. Acad. Sci. U.S.A. 74:3480–3484.
Spandidos, D., and Siminovitch, L. (1977).Cell 12:235–242.
Spandidos, D., and Siminovitch, L. (1977).Cell 12:675–682.
Spandidos, D., and Siminovitch, L. (1977).Brookhaven Symposium Biology 29:127–134.
Spandidos, D., and Siminovitch, L. (1978).Nature 271:259–261.
Kit, S., Dubbs, D., Piekarski, L., and Hsu, T. (1963).Exp. Cell Res. 31:291–312.
McBurney, M. W., and Whitmore, G. F. (1974).Cell 2:173–182.
Flintoff, W. F., Davidson, S. V., and Siminovitch, L. (1976).Somat. Cell Genet. 2:245–261.
Stanners, C. P., Elicieri, G. L., and Green, H. (1971).Nature (London) 230:52–54.
Willecke, K., and Ruddle, F. H. (1975).Proc. Natl. Acad. Sci. U.S.A. 72:1792–1796.
Burton, K. (1956).Biochem. J. 62:315–323.
Pellicer, A., Wigler, M., Axel, R., and Silverstein, S. (1978).Cell 14:133–141.
Graham, F. L., and van der Eb, A. J. (1973).Virology 52:456–567.
Srinivasan, P. R., and Lewis, W. H. (1980). InIntroduction of Macromolecules into Viable Mammalian Cells (ed.) Baserga, R., Croce, C., and Rovera, G. (Alan R. Liss, New York), Wistar Symposium Series1:27–45.
Lowry, O. H., Rosebrough, N. J., Farr, A. L., and Randall, R. J. (1951).J. Biol. Chem. 193:265–275.
Friend, C., Scher, W., Holland, J. G., and Sato, T. (1971).Proc. Natl. Acad. Sci. U.S.A. 68:378–382.
Terada, M., Fried, J., Nudel, V., Rifkind, R. A., and Marks, P. (1977).Proc. Natl. Acad. Sci. U.S.A. 74:248–252.
Scher, W., and Friend, C. (1978).Cancer Res. 38:841–849.
Terada, M., Nudel, U., Fibach, E., Rifkind, R. A., and Marks, P. (1978).Cancer Res. 38:835–840.
Stow, N. D., and Wilkie, N. M. (1976).J. Gen. Virol. 33:447–458.
Norwood, T. H., Ziegler, C. J., and Martin, C. M. (1976).Somat. Cell Genet. 2:263–270.
Lyman, G. H., Preisler, H. D., and Papahadjopoulos, D. (1976).Nature 262:360–363.
Lewis, W. H., and Wright, J. A. (1978).J. Cell. Physiol. 97:87–98.
Lewis, W. H., and Wright, J. A. (1979).Somat. Cell Genet. 5:83–96.
Thompson, L., Harkins, J., and Stanners, C. (1973).Proc. Natl. Acad. Sci. U.S.A. 70:3094–3098.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Lewis, W.H., Srinivasan, P.R., Stokoe, N. et al. Parameters governing the transfer of the genes for thymidine kinase and dihydrofolate reductase into mouse cells using metaphase chromosomes or DNA. Somat Cell Mol Genet 6, 333–348 (1980). https://doi.org/10.1007/BF01542787
Received:
Revised:
Issue Date:
DOI: https://doi.org/10.1007/BF01542787