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首页> 外文期刊>Molecular and Cellular Biology >Stable gene amplification and overexpression of sodium- and potassium-activated ATPase in HeLa cells.
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Stable gene amplification and overexpression of sodium- and potassium-activated ATPase in HeLa cells.

机译：HeLa细胞中稳定的基因扩增和钠和钾激活的ATPase的过表达。

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Cell lines stably resistant to ouabain were isolated from an unstably resistant HeLa line after growth in nonselective medium. Stable resistant lines bound ouabain at levels 10-fold higher than did HeLa cells and at similar levels to those bound by the unstable C+ line previously described (J. F. Ash, R. M. Fineman, T. Kalka, M. Morgan, and B. Wire, J. Cell Biol. 99: 971-983). Expression and synthesis of the Na+, K+ -ATPase alpha chain showed a similar amplification over that for HeLa cells by Western blots and [35S]methionine pulse-labeling. In addition, a glycoprotein labeled with [3H]fucose and comigrating with the Na+, K+ -ATPase beta chain was eight- to ninefold amplified in stably resistant lines. Dot blots with a cDNA clone specific for Na+, K+ -ATPase alpha chain gene sequences confirmed the amplification of this gene. Karyotyping suggested that the amplification is associated with an expanded, abnormal banded region on the long (q) arm of one chromosome 17.
机译：在非选择性培养基中生长后，从不稳定的HeLa细胞系中分离出对哇巴因具有稳定抗性的细胞系。稳定的抗性系结合哇巴因的水平比HeLa细胞高10倍，并且与先前描述的不稳定C +系结合的水平相似（JF Ash，RM Fineman，T。Kalka，M。Morgan和B. Wire，J （Cell Biol.99：971-983）。通过蛋白质印迹和[35S]蛋氨酸脉冲标记，Na +，K + -ATPaseα链的表达和合成显示出与HeLa细胞相似的扩增。另外，用[3H]岩藻糖标记并与Na +，K + -ATPaseβ链竞争的糖蛋白在稳定抗性品系中扩增了八至九倍。用对Na +，K + -ATPaseα链基因序列具有特异性的cDNA克隆进行斑点印迹，证实了该基因的扩增。核型分析表明，扩增与一个染色体17的长（q）臂上的扩大的异常条带区域相关。

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