Genetic transformation of Drosophila cells in culture by P element-mediated transposition

Daniel Segal, Lucy Cherbas, Peter Cherbas

Research output: Contribution to journalArticlepeer-review

Abstract

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 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 using Drosophila cells.

Original languageEnglish
Pages (from-to)159-165
Number of pages7
JournalSomatic Cell and Molecular Genetics
Volume22
Issue number2
DOIs
StatePublished - Mar 1996

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