A general strategy for selecting high-affinity zinc finger proteins for diverse DNA target sites
A general strategy for selecting high-affinity zinc finger proteins for diverse DNA target sites
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DOI：
10.1126/science.275.5300.657
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发表时间：
1997-01-31
期刊：
影响因子：
56.9
通讯作者：
Pabo, CO
中科院分区：
文献类型：
--
作者：
Greisman, HA;Pabo, CO
描述了一种选择识别所需序列的dna结合蛋白的方法。该方案包括在所需的9或10个碱基对靶点上逐渐延伸一个新的锌指蛋白，每次添加和优化一个锌指。该过程用TATA盒、p53结合位点和核受体元件进行了测试，获得了与纳摩尔解离常数结合并有效（大于20,000倍）区分非特异性DNA的蛋白质。这一策略可能为蛋白质- dna识别提供重要信息，并为生物医学研究提供有力工具。
A method is described for selecting DNA-binding proteins that recognize desired sequences. The protocol involves gradually extending a new zinc finger protein across the desired 9- or 10-base pair target site, adding and optimizing one finger at a time. This procedure was tested with a TATA box, a p53 binding site, and a nuclear receptor element, and proteins were obtained that bind with nanomolar dissociation constants and discriminate effectively (greater than 20,000-fold) against nonspecific DNA. This strategy may provide important information about protein-DNA recognition as well as powerful tools for biomedical research.