Abstract
A MoMLV-based retroviral vector capable of transmitting and expressing both the human hypoxanthine phosphoribosyltransferase (hprt)coding sequence and the Herpes simplex type 1 thymidine kinase (tk)gene has been constructed. After infection of a rat cell line, cell clones were selected on the basis of expressing both markers. They were subsequently found to contain a single provirus of the expected topology. The ease with which loss of expression of the markers can be monitored has allowed us to make observations on the stability of proviral genes. In particular, we have found indirect evidence of strong position effects on proviral gene expression by comparing the characteristic frequency of marker loss in different clonal proviral lines. Effects of the selection protocol on the apparent frequency of variants have also been noted. Finally, a combination of molecular and genetic observations lead us to invoke chromosome loss as the major factor influencing marker stability in this system.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
Literature cited
Cepko, C.L., Roberts, B.E., and Mulligan, R.C. (1984).Cell 37:1053.
Mann, R., Mulligan, R.C., and Baltimore, D. (1983).Cell 33:153.
Varmus, H.E., Quintrell, N., and Ortiz, S. (1981).Cell 25:23.
Varmus, H.E., Quintrell, N., and Wyke, J. (1981).Virology 108:28.
Hughes, S.H., Shank, P.R., Spector, D.H., Kung, H.-J., Bishop, J.M., Vogt, P.K., and Breitman, M.L. (1978).Cell 15:1397.
Steffan, D., and Weinberg, R.A. (1978).Cell 15:1003.
Jolly, D.J., Willis R.C., and Friedmann, T. (1986).Mol. Cell. Biol. 6:1141.
Stanners, C.P., Elicien, G.L., and Green, H. (1971).Nature (London), New Biol. 230:52.
Topp, W.C. (1981).Virology 113:408.
Quade, K. (1979).Virology 98:461.
Tabin, C.J., Hoffmann, J.W., Goff, S.P., and Weinberg, R.A. (1982).Mol. Cell. Biol. 2:426.
Miller, A.D., Jolly, D.J., Friedmann, T., and Verma, I.M. (1983).Proc. Natl. Acad. Sci. U.S.A. 80:4709.
Harbers, K., Schnieke, A., Stuhlmann, H., Jahner, D., and Jaenisch, R. (1978).Proc. Natl. Acad. Sci. U.S.A. 78:7609.
Dunn, R.J., Belagaje, R., Brown, E.L., and Khorana H.G. (1981).J. Biol. Chem. 256:6109.
Southern, P.J., and Berg, P. (1982).J. Mol. Appl. Genet. 1:327.
Graham, F.L., Bacchetti, S., and Mckinnon, R., Stanners, C., Cordell, B., and Goodman, H.M. (1980). In Baserga, R., Croce, C., and Rprera, G. (eds.),Introduction of Macromolecules into Viable Mammalian Cells. (Alan R. Liss, New York), pp. 3–25.
Perucho, M., Goldfarb, M., Shimuzu, K., Larna, C., Fogh, J., and Wigler, M. (1981).Cell 27:467.
Meinkoth, J., and Wahl, G. (1984).Anal. Biochem. 138:267.
Sutter, D., and Doerfler, W. (1980).Proc. Natl. Acad. Sci. U.S.A. 77:253.
Jones, P.A., and Taylor, S.M. (1980).Cell 20:85.
Varmus, H.E. (1982).Science 216:812.
Groudine, M., Eisenman, R., and Weintraub, H. (1981).Nature 292:311.
Emerman, M., and Temin, H.M. (1984).Cell 39:459.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Stone, J.C., Dower, N.A. & Siminovitch, L. Stability of retrovirally transduced markers in a rat cell line. Somat Cell Mol Genet 12, 575–583 (1986). https://doi.org/10.1007/BF01671943
Received:
Revised:
Issue Date:
DOI: https://doi.org/10.1007/BF01671943