Burke J F, Pinchin S M, Ish-Horowicz D, Sinclair J H, Sang J H
Somat Cell Mol Genet. 1984 Nov;10(6):579-88. doi: 10.1007/BF01535223.
We have used DNA-mediated gene transfer to introduce into Drosophila melanogaster cells DNA sequences for which no selective criteria exist. We have introduced a Drosophila heat-shock locus into cultured Drosophila cells by calcium phosphate cotransfection with the copia vector pCV31gpt and selection for xanthine utilization. We recovered cell lines containing between three and about 50 copies of both pCV31gpt and pMH10A, a cloned 87 A7 hsp70 heat-shock locus that encodes a mutant 40,000-dalton heat-shock protein (hsp40). The stable inheritance of the transformed DNAs argues that the input DNAs have integrated into the genome. We show that this is indeed the case for one cell line by cloning back the transfected DNA and detecting the flanking chromocentral sequences by in situ hybridization. Surprisingly, the integrated hsp70 genes are not expressed. This report represents the first example of the cointroduction of DNA sequences into Drosophila cells by cotransfection with a dominant selectable marker.
我们利用DNA介导的基因转移技术,将不存在选择标准的DNA序列导入黑腹果蝇细胞。我们通过磷酸钙共转染,将果蝇热休克基因座与copia载体pCV31gpt一同导入培养的果蝇细胞,并通过黄嘌呤利用进行筛选。我们获得了含有3至约50个拷贝的pCV31gpt和pMH10A的细胞系,pMH10A是一个克隆的87A7 hsp70热休克基因座,编码一种突变的40,000道尔顿热休克蛋白(hsp40)。转化DNA的稳定遗传表明输入的DNA已整合到基因组中。我们通过克隆回转染的DNA并通过原位杂交检测侧翼着丝粒序列,证明了一个细胞系确实如此。令人惊讶的是,整合的hsp70基因不表达。本报告代表了通过与显性选择标记共转染将DNA序列共导入果蝇细胞的首个实例。