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构建一种赋予培养中的哺乳动物细胞对潮霉素B抗性的融合基因。

Construction of a fusion gene that confers resistance against hygromycin B to mammalian cells in culture.

作者信息

Bernard H U, Krämmer G, Röwekamp W G

出版信息

Exp Cell Res. 1985 May;158(1):237-43. doi: 10.1016/0014-4827(85)90446-x.

Abstract

Mouse L fibroblasts and other mammalian cells are killed by the translation inhibitor hygromycin B. We have modified the gene conferring resistance against hygromycin B in E. coli in such a way that it can be transcribed in mammalian cells from the promoter of the HSVtk gene. The resulting plasmid, pHMR272, was transfected into mouse L fibroblasts and HeLa cells by the calcium phosphate method and upon selection produced clones resistant against hygromycin B. The transfection rate was similar to that obtained with other selective markers. This plasmid is a useful addition to the relatively small number of dominant selectable markers available for mammalian cells.

摘要

小鼠L成纤维细胞和其他哺乳动物细胞会被翻译抑制剂潮霉素B杀死。我们对大肠杆菌中赋予潮霉素B抗性的基因进行了改造,使其能够在哺乳动物细胞中由单纯疱疹病毒胸苷激酶(HSVtk)基因的启动子转录。将得到的质粒pHMR272通过磷酸钙法转染到小鼠L成纤维细胞和HeLa细胞中,经筛选产生了对潮霉素B有抗性的克隆。转染率与使用其他选择标记获得的转染率相似。该质粒是对可用于哺乳动物细胞的相对较少的显性选择标记的有益补充。

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