通过基因靶向在人类细胞中构建一个能使其DNA重新复制的“条件性”细胞周期蛋白依赖性激酶2（CDC2）突变体。
Construction by gene targeting in human cells of a "conditional' CDC2 mutant that rereplicates its DNA.
作者信息
Itzhaki J E, Gilbert C S, Porter A C
机构信息
MRC Clinical Sciences Centre, Royal Postgraduate Medical School, Hammersmith Hospital, London, UK.
出版信息
Nat Genet. 1997 Mar;15(3):258-65. doi: 10.1038/ng0397-258.
We describe a novel gene targeting strategy for the genetic analysis of essential genes in mammalian cells and its use to study the role of the cell cycle control gene CDC2 in human cells. A cell line (HT2-19) was generated in which endogenous CDC2 gene expression and cell viability depend on the presence of an inducer in the growth medium. In the absence of inducer, HT2-19 cells undergo extensive DNA rereplication and apoptosis. Rereplication is indicative of a role for human CDC2 in a control mechanism, previously undetected in mammalian cells, that prevents premature entry into S-phase.
我们描述了一种用于哺乳动物细胞中必需基因遗传分析的新型基因靶向策略，以及该策略用于研究细胞周期控制基因CDC2在人类细胞中的作用。构建了一种细胞系（HT2-19），其中内源性CDC2基因的表达和细胞活力取决于生长培养基中诱导剂的存在。在没有诱导剂的情况下，HT2-19细胞会经历广泛的DNA再复制和凋亡。再复制表明人类CDC2在一种以前在哺乳动物细胞中未被发现的控制机制中发挥作用，该机制可防止过早进入S期。
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