Selection of highly transfectable variant from mouse mastocytoma P815
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A tk− cell line derived from mouse mastocytoma P815 was transfected with a plasmid carrying a thymidine kinase gene. The tk+ cells were obtained at a frequency of 10−6. From some of these tk+ transfectants it was possible to select tk− cells with BrdU so that these cells could be submitted again to transfection with the thymidine kinase gene. By repeating cycles of transfection and tk+ selection followed by reverse selection with BrdU, it was possible to obtain a stable variant having a 100-fold increase in transfection efficiency. This HTR (high transfection) variant shows a high efficiency of transfection (10−4) for the neomycin-resistance gene as well as for the thymidine kinase gene.
KeywordsThymidine Transfection Efficiency Thymidine Kinase Kinase Gene Stable Variant
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- 21.Tonkonogy, S.L., and Amos, D.B. (1981).Immuno-genetics 14:497–505.Google Scholar
- 25.Nicolas, J.F., and Berg, P. (1983). Teratocarcinoma stem cells. Cold Spring Harbor Conferences on Cell Proliferation, Vol. 10, pp. 469–485.Google Scholar
- 26.Davis, R.W., Botstein, D., and Roth, J.R. (1980). InA Manual for Genetic Engineering. (Cold Spring Harbor Laboratory, Cold Spring Harbor, New York), pp. 116–119.Google Scholar
- 39.Maryanski, Y.L., Moretta, A., Jordan, B., De Plaen, E., Van Pel, A., Boon, T. and Cerottini, Y-C. (1985)Eur. Y. Immunol., in press.Google Scholar