Matthias P D, Bernard H U, Scott A, Brady G, Hashimoto-Gotoh T, Schütz G
Institute of Cell and Tumor Biology, German Cancer Research Center, Heidelberg, FRG.
EMBO J. 1983;2(9):1487-92. doi: 10.1002/j.1460-2075.1983.tb01612.x.
We describe the construction of a bovine papilloma virus-based vector (pCGBPV9) which contains a dominant selectable marker and replicates autonomously in both mouse and Escherichia coli cells. This vector contains the complete bovine papilloma virus genome, a ColE1 replication origin and a dominant selectable marker conferring resistance to kanamycin in bacteria and G418 in eukaryotic cells. A high number of G418R colonies are obtained after transfer of pCGBPV9 into mouse C127 cells. These G418R colonies contain vector DNA which replicates autonomously at approximately 10-30 copies per cell. The molecules are in most cases unrearranged and can be rescued into E. coli cells by bacterial transformation.
我们描述了一种基于牛乳头瘤病毒的载体(pCGBPV9)的构建,该载体包含一个显性选择标记,并能在小鼠细胞和大肠杆菌细胞中自主复制。此载体包含完整的牛乳头瘤病毒基因组、一个ColE1复制起点以及一个显性选择标记,该标记赋予细菌对卡那霉素和真核细胞对G418的抗性。将pCGBPV9转入小鼠C127细胞后获得了大量G418抗性菌落。这些G418抗性菌落含有载体DNA,其在每个细胞中以约10 - 30个拷贝的数量自主复制。在大多数情况下,这些分子未发生重排,并且可以通过细菌转化拯救到大肠杆菌细胞中。