Rescue of a Drosophila temperature-sensitive mutant cell line by DNA transfection

At the nonpermissive temperature, the shibirets gene of Drosophila causes adult paralysis and cell death during development. We have used DNA-mediated gene transfer with wild-type genomic DNA to rescue the lethal phenotype of shits cultured cells and have isolated cell lines which grow at the nonpermissive temperature of 29 degrees C. By using restriction endonucleases, we confirm that this rescue is DNA sequence dependent. Further, cells cotransfected with wild-type genomic DNA and plasmid DNA contain plasmid sequences when selected for survival at 29 degrees C. This confirms the uptake of DNA by the transformed cells and provides a simple temperature selection technique allowing coselection of genes which cannot themselves be selected for.