European Molecular Biology Laboratory, Postfach 10.2209, 6900 Heidelberg, FRG.
EMBO J. 1985 Jan;4(1):167-71. doi: 10.1002/j.1460-2075.1985.tb02332.x.
Drosophila larvae are rapidly killed by food containing the antibiotic G418. The bacterial gene for neomycin resistance introduced in the genome by P-mediated transformation renders larvae resistant to G418 and able to grow to fertile adults. The neo gene transcribed from the herpes thymidine kinase promoter gives low levels of resistance but high levels can be obtained using the hsp70 heat-shock promoter. We have constructed a vector for P-mediated transformation which uses this finding to allow dominant selection of transformed progeny. Features of this vector also facilitate cloning and allow the rapid recovery of the inserted transposon from transformed flies. We have also constructed a cosmid vector for P-mediated transformation that incorporates the hsp70-neo gene.
果蝇幼虫会因摄入含抗生素 G418 的食物而迅速死亡。通过 P 介导的转化引入基因组的新霉素抗性细菌基因使幼虫对 G418 具有抗性,并能够生长为有生育能力的成虫。由单纯疱疹胸苷激酶启动子转录的 neo 基因赋予低水平的抗性,但使用 hsp70 热休克启动子可以获得高水平的抗性。我们构建了一个用于 P 介导转化的载体,该载体利用这一发现允许转化后代的显性选择。该载体的特点还便于克隆,并允许从转化的果蝇中快速回收插入的转座子。我们还构建了一个用于 P 介导转化的 cosmid 载体,其中包含 hsp70-neo 基因。