Genetic transformation of Drosophila cells in culture by P element-mediated transposition.
We report that cells of a Drosophila 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 alpha-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 (< or = 5%) retain plasmid sequences derived from illegitimate recombination. Using conditions like these it should prove possible to construct enhancer trap and/or gene libraries using Drosophila cells.
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Department of Molecular Microbiology and Biotechnology, Tel-Aviv University, Israel.
Cherbas
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Cherbas
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D000546:Amanitins / Q000494:pharmacology; D000818:Animals; D002461:Cell Line, Transformed; D004247:DNA / Q000032:analysis; D004254:DNA Nucleotidyltransferases / Q000235:genetics; D004251:DNA Transposable Elements; D004330:Drosophila / Q000166:cytology* / Q000235:genetics; D004351:Drug Resistance; D004625:Embryo, Nonmammalian; D005493:Folic Acid Antagonists / Q000494:pharmacology; D005822:Genetic Vectors / Q000235:genetics; D008727:Methotrexate / Q000494:pharmacology; D013762:Tetrahydrofolate Dehydrogenase / Q000235:genetics; D014162:Transfection; D014170:Transformation, Genetic; D019895:Transposases
D000546:Amanitins; D004251:DNA Transposable Elements; D005493:Folic Acid Antagonists; D004247:DNA; D013762:Tetrahydrofolate Dehydrogenase; D004254:DNA Nucleotidyltransferases; D019895:Transposases; D008727:Methotrexate