Sequence-Defined Scaffolding of Peptides on Nucleic Acid Polymers
Sequence-Defined Scaffolding of Peptides on Nucleic Acid Polymers
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DOI：
10.1021/jacs.5b07675
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发表时间：
2015-09-02
影响因子：
15
通讯作者：
Hili, Ryan
中科院分区：
文献类型：
--
作者：
Guo, Chun;Watkins, Christopher P.;Hili, Ryan
我们发展了一种T4 DNA连接酶催化的DNA模板聚合含5 ′-磷酸化五核苷酸的肽段的方法。聚合进行序列特异性地以优异的产率产生DNA支架肽。该方法已被证明耐受长度为2至8个氨基酸的肽，具有各种各样的功能。我们验证了该系统在模拟选择中富集来自文库的His标记的DNA支架肽表型的能力，该文库在一轮选择后表现出190倍的富集。该策略展示了一种有前途的新方法，允许基于肽片段的单链DNA支架的高亲和力试剂的生成和体外选择。
We have developed a method for the T4 DNA ligase-catalyzed DNA-templated polymerization of 5'-phosphorylated pentanucleotides containing peptide fragments. The polymerization proceeds sequence-specifically to generate DNA-scaffolded peptides in excellent yields. The method has been shown to tolerate peptides ranging from two to eight amino acids in length with a wide variety of functionality. We validated the capabilities of this system in a mock selection for the enrichment of a His-tagged DNA-scaffolded peptide phenotype from a library, which exhibited a 190-fold enrichment after one round of selection. This strategy demonstrates a promising new approach to allowing the generation and in vitro selection of high-affinity reagents based upon single-stranded DNA scaffolding of peptide fragments.