Abstract
We report that cells of aDrosophila embryonic cell line (Kc167 cells) can be readily and stably transformed by transposition of P elements from exogenous DNA. Cells are transfected with plasmids carrying methotrexate- or α-amanitin-resistance markers expressed from constitutive promoters and co-transfected with a gene encoding a somatically active transposase. Transient expression of the transposase leads to efficient production of transformed, resistant cells. We describe conditions under which most resistant clones are healthy and harbor a small number (1–50) of transposons and few (≤5%) retain plasmid sequences derived from illegitimate recombination. Using conditions like these it should prove possible to construct enhancer trap and/or gene libraries usingDrosophila cells.
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Segal, D., Cherbas, L. & Cherbas, P. Genetic transformation ofDrosophila cells in culture by P element-mediated transposition. Somat Cell Mol Genet 22, 159–165 (1996). https://doi.org/10.1007/BF02369906
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DOI: https://doi.org/10.1007/BF02369906