Tuerk C, Gold L
Department of Molecular, Cellular, Developmental Biology, University of Colorado, Boulder 80309.
Science. 1990 Aug 3;249(4968):505-10. doi: 10.1126/science.2200121.
High-affinity nucleic acid ligands for a protein were isolated by a procedure that depends on alternate cycles of ligand selection from pools of variant sequences and amplification of the bound species. Multiple rounds exponentially enrich the population for the highest affinity species that can be clonally isolated and characterized. In particular one eight-base region of an RNA that interacts with the T4 DNA polymerase was chosen and randomized. Two different sequences were selected by this procedure from the calculated pool of 65,536 species. One is the wild-type sequence found in the bacteriophage mRNA; one is varied from wild type at four positions. The binding constants of these two RNA's to T4 DNA polymerase are equivalent. These protocols with minimal modification can yield high-affinity ligands for any protein that binds nucleic acids as part of its function; high-affinity ligands could conceivably be developed for any target molecule.
通过一种依赖于从变异序列库中交替进行配体选择和结合物种扩增的程序，分离出了针对一种蛋白质的高亲和力核酸配体。多轮操作以指数方式富集群体中具有最高亲和力的物种，这些物种可进行克隆分离和表征。特别地，选择了与T4 DNA聚合酶相互作用的RNA的一个八碱基区域并使其随机化。通过该程序从计算出的65536种序列库中选择了两种不同的序列。一种是在噬菌体mRNA中发现的野生型序列；一种在四个位置上与野生型不同。这两种RNA与T4 DNA聚合酶的结合常数相当。这些方案只需进行最少的修改，就能为任何作为其功能一部分而结合核酸的蛋白质产生高亲和力配体；可以想象，针对任何靶分子都能开发出高亲和力配体。
