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用显性选择标记对培养的黑腹果蝇细胞进行转化。

Transformation of cultured Drosophila melanogaster cells with a dominant selectable marker.

作者信息

Rio D C, Rubin G M

出版信息

Mol Cell Biol. 1985 Aug;5(8):1833-8. doi: 10.1128/mcb.5.8.1833-1838.1985.

Abstract

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.

摘要

我们已经开发出一种方法,可将外源DNA稳定且高效地导入黑腹果蝇组织培养细胞。构建了一种质粒载体,其携带在copia转座元件长末端重复启动子转录控制下的细菌新霉素抗性基因。经磷酸钙-DNA转染后,该载体使黑腹果蝇细胞对庆大霉素衍生物氨基糖苷类G-418产生抗性。载体DNA似乎以每个细胞10至20个拷贝的长串联阵列形式整合,并且在无选择条件下可稳定存在许多代。为了测试该系统将未选择的DNA导入黑腹果蝇细胞的实用性,将P转座元件与hsp70启动子之间的基因融合体插入到copia-新霉素抗性质粒中。转染并建立G-418抗性细胞系后,发现热休克后hsp-P融合基因能高效转录。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e424/366898/984391849eae/molcellb00104-0035-a.jpg

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