SYSTEMATIC EVOLUTION OF LIGANDS BY EXPONENTIAL ENRICHMENT - RNA LIGANDS TO BACTERIOPHAGE-T4 DNA-POLYMERASE
SYSTEMATIC EVOLUTION OF LIGANDS BY EXPONENTIAL ENRICHMENT - RNA LIGANDS TO BACTERIOPHAGE-T4 DNA-POLYMERASE
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DOI：
10.1126/science.2200121
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发表时间：
1990-08-03
期刊：
影响因子：
56.9
通讯作者：
GOLD, L
中科院分区：
文献类型：
--
作者：
TUERK, C;GOLD, L
蛋白质的高亲和力核酸配体通过依赖于从变体序列池中选择配体和扩增结合物质的交替循环的程序分离。多轮以指数方式富集群体中可克隆分离和表征的最高亲和力物种。特别地，选择并随机化与T4 DNA聚合酶相互作用的RNA的一个八碱基区域。通过该程序从计算的65，536个物种的库中选择两个不同的序列。一个是在噬菌体mRNA中发现的野生型序列;一个在四个位置与野生型不同。这两种RNA与T4 DNA聚合酶的结合常数是相等的。这些具有最小修改的方案可以为任何结合核酸作为其功能的一部分的蛋白质产生高亲和力配体;可以想象，可以为任何靶分子开发高亲和力配体。
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.