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带有显性抗性标记的牛乳头瘤病毒载体在小鼠和大肠杆菌细胞中进行染色体外复制。

A bovine papilloma virus vector with a dominant resistance marker replicates extrachromosomally in mouse and E. coli cells.

作者信息

Matthias P D, Bernard H U, Scott A, Brady G, Hashimoto-Gotoh T, Schütz G

机构信息

Institute of Cell and Tumor Biology, German Cancer Research Center, Heidelberg, FRG.

出版信息

EMBO J. 1983;2(9):1487-92. doi: 10.1002/j.1460-2075.1983.tb01612.x.

Abstract

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.

摘要

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。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/37e5/555311/c91ab76e5b0e/emboj00262-0061-a.jpg

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