Desjarlais J R, Berg J M
Thomas C. Jenkins Department of Biophysics, Johns Hopkins University, Baltimore, MD 21218.
Proc Natl Acad Sci U S A. 1994 Nov 8;91(23):11099-103. doi: 10.1073/pnas.91.23.11099.
The screening of combinatorial libraries is becoming a powerful method for identifying or refining the structures of ligands for binding proteins, enzymes, and other receptors. We describe an oligonucleotide library search procedure in which the identity of each member is encoded in the length of oligonucleotides. This encoding scheme allows binding-site preferences to be evaluated via DNA length determination by denaturing gel electrophoresis. We have applied this method to determine the binding-site preferences for 18 Cys2His2 zinc finger domains as the central domain within a fixed context of flanking zinc fingers. An advantage of the method is that the relative affinities of all members of the library can be estimated in addition to simply determining the sequence of the optimal or consensus ligand. The zinc finger domain specificities determined will be useful for modular zinc finger protein design.
组合文库的筛选正成为一种强大的方法，用于识别或优化与结合蛋白、酶及其他受体结合的配体结构。我们描述了一种寡核苷酸文库搜索程序，其中每个成员的身份由寡核苷酸的长度编码。这种编码方案允许通过变性凝胶电泳测定DNA长度来评估结合位点偏好。我们已应用此方法确定了18个Cys2His2锌指结构域在侧翼锌指的固定背景下作为中心结构域的结合位点偏好。该方法的一个优点是，除了简单地确定最佳或共有配体的序列外，还可以估计文库中所有成员的相对亲和力。所确定的锌指结构域特异性将有助于模块化锌指蛋白的设计。
