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首页> 外文期刊>Journal of Molecular Biology >PROMOTER-SPECIFIC ACTIVATION OF GENE EXPRESSION DIRECTED BY BACTERIOPHAGE-SELECTED ZINC FINGERS
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PROMOTER-SPECIFIC ACTIVATION OF GENE EXPRESSION DIRECTED BY BACTERIOPHAGE-SELECTED ZINC FINGERS

机译：噬菌体选择的锌指指示基因表达的启动子特异性激活。

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摘要

It has been shown that sequence-specific DNA-binding domains containing zinc fingers can be selected from libraries displayed on filamentous bacteriophage. The affinity and specificity of these peptides are well characterised in vitro, but few data are available to demonstrate specific DNA binding and discrimination between closely related DNA sequences in vivo. Transient transactivation assays were performed in mammalian cells, using expression plasmids which produce different amounts of a model transcription factor containing a phage-selected zinc finger DNA-binding domain, and reporter plasmids which carry systematic variations of the promoter sequence. When the intracellular concentration of the transcription factor was appropriate, activation of gene expression was absolutely dependent on a promoter having the same DNA sequence as that originally used to select the zinc finger domain by phage display. However, excessive intracellular concentrations of the transcription factor resulted in some less-specific DNA binding, leading to gene activation from similar promoters containing a maximum of two base changes. Thus, provided delivery is carefully controlled, highly specific control of gene expression in vivo can be achieved using artificial transcription factors containing phage-selected zinc finger DNA-binding domains. (C) 1997 Academic Press Limited. [References: 35]
机译：已经显示，可以从在丝状噬菌体上展示的文库中选择含有锌指的序列特异性DNA结合结构域。这些肽的亲和力和特异性在体外已得到很好的表征，但很少有数据可证明特异性DNA结合和体内密切相关的DNA序列之间的区别。在哺乳动物细胞中进行瞬时反式激活测定，使用产生不同量的包含噬菌体选择的锌指DNA结合结构域的模型转录因子的表达质粒和携带启动子序列的系统变异的报告质粒。当转录因子的细胞内浓度合适时，基因表达的激活绝对取决于具有与最初用于通过噬菌体展示选择锌指结构域的DNA序列相同的DNA序列的启动子。但是，转录因子的细胞内浓度过高会导致一些特异性较低的DNA结合，从而导致包含最多两个碱基变化的类似启动子的基因活化。因此，只要小心控制递送，就可以使用含有噬菌体选择的锌指DNA结合结构域的人工转录因子来实现体内基因表达的高度特异性控制。 （C）1997 Academic Press Limited。 [参考：35]

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