Transformation of cultured Drosophila melanogaster cells with a dominant selectable marker.
We have developed a method for the stable and efficient introduction of foreign DNA into Drosophila melanogaster tissue culture cells. A plasmid vector was constructed that carries the bacterial neomycin resistance gene under the transcriptional control of the copia transposable element long terminal repeat promoter. After calcium phosphate-DNA transfection, this vector rendered D. melanogaster cells resistant to the aminoglycoside G-418, a derivative of gentamicin. The vector DNA appeared to be integrated in long tandem arrays of 10 to 20 copies per cell and was stable for many generations in the absence of selection. To test the usefulness of this system for introducing nonselected DNA into D. melanogaster cells, a gene fusion between the P transposable element and the hsp70 promoter was inserted into the copia-neomycin resistance plasmid. After transfection and establishment of a G-418-resistant cell line, the hsp-P fusion gene was found to be efficiently transcribed after heat shock.
D000818:Animals; D001419:Bacteria / Q000235:genetics; D002460:Cell Line; D004262:DNA Restriction Enzymes; D004251:DNA Transposable Elements; D004331:Drosophila melanogaster / Q000235:genetics*; D004352:Drug Resistance, Microbial; D005799:Genes, Dominant; D005822:Genetic Vectors; D009355:Neomycin / Q000494:pharmacology; D011815:R Factors; D014158:Transcription, Genetic; D014170:Transformation, Genetic
D004251:DNA Transposable Elements; D004262:DNA Restriction Enzymes; D009355:Neomycin