Discovery, characterization, and optimization of an unnatural base pair for expansion of the genetic alphabet
Discovery, characterization, and optimization of an unnatural base pair for expansion of the genetic alphabet
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DOI：
10.1021/ja078223d
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发表时间：
2008-02-20
影响因子：
15
通讯作者：
Romesberg, Floyd E.
中科院分区：
文献类型：
--
作者：
Leconte, Aaron M.;Hwang, Gil Tae;Romesberg, Floyd E.
DNA固有地受到其四种天然核酸的限制。通过添加非自然碱基对来扩展遗传字母表的努力，有望扩大DNA的生物技术应用，也是扩展遗传密码的重要第一步。我们已经进行了两个独立的疏水性非天然核苷酸筛选，以确定新的候选碱基对，以及天然DNA聚合酶识别。从3600个候选碱基对的池中，两个筛选鉴定出相同的碱基对，dSICS：dMMO 2，我们在这里报告。使用一系列相关的类似物，我们进行了详细的结构-活性关系分析，这使我们能够确定每个核碱基上的必要官能团。从这些研究的结果，我们设计了一个优化的碱基对，d5 SICS：dMMO 2，这是有效的和选择性合成的KF内的背景下，天然DNA。
DNA is inherently limited by its four natural nuclectides. Efforts to expand the genetic alphabet, by addition of an unnatural base pair, promise to expand the biotechnological applications available for DNA as well as to be an essential first step toward expansion of the genetic code. We have conducted two independent screens of hydrophobic unnatural nucleotides to identify novel candidate base pairs that are well recognized by a natural DNA polymerase. From a pool of 3600 candidate base pairs, both screens identified the same base pair, dSICS:dMMO2, which we report here. Using a series of related analogues, we performed a detailed structure-activity relationship analysis, which allowed us to identify the essential functional groups on each nucleobase. From the results of these studies, we designed an optimized base pair, d5SICS:dMMO2, which is efficiently and selectively synthesized by Kf within the context of natural DNA.