A general strategy for selecting high-affinity zinc finger proteins for diverse DNA target sites

被引：310

作者：

Greisman, HA

Pabo, CO

机构：

[1] MIT,HOWARD HUGHES MED INST,CAMBRIDGE,MA 02139

[2] MIT,DEPT BIOL,CAMBRIDGE,MA 02139

来源：

SCIENCE | 1997年 / 275卷 / 5300期

关键词：

D O I：

10.1126/science.275.5300.657

中图分类号：

O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];

学科分类号：

07 ; 0710 ; 09 ;

摘要：

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.

引用

页码：657 / 661

页数：5

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