Enzymatic Synthesis, Amplification, and Application of DNA with a Functionalized Backbone,Angewandte Chemie International Edition

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Enzymatic Synthesis, Amplification, and Application of DNA with a Functionalized Backbone
Angewandte Chemie International Edition ( IF 16.1 ) Pub Date : 2017-10-10 , DOI: 10.1002/anie.201707367
Tingjian Chen ₁ , Floyd E. Romesberg ₁

Affiliation

The ability to amplify DNA along with its unprecedented sequence control has led to its use for different applications, but all are limited by the properties available to natural nucleotides. We previously reported the evolution of polymerase SFM4‐3, which better tolerates 2′‐modified substrates. To explore the utility of SFM4‐3, we now report the characterization of its recognition of substrates with 2′‐azido, 2′‐chloro, 2′‐amino, or arabinose sugars. We find that SFM4‐3 can efficiently synthesize polymers composed of these nucleotides, and most interestingly, that SFM4‐3 can also PCR amplify these modified oligonucleotides. When combined with post‐amplification modification, the latter allows for the exponential amplification of polymers that may be functionalized with desired moieties arrayed in a controlled fashion, the utility of which we demonstrate with extensive small molecule functionalization and the production and initial characterization of a novel DNA hydrogel.

中文翻译：

酶的合成，扩增和功能化骨干DNA的应用。

扩增DNA的能力及其空前的序列控制已导致其可用于不同的应用，但是所有这些都受到天然核苷酸可利用的特性的限制。我们之前曾报道过聚合酶SFM4-3的进化过程，它可以更好地耐受2'-修饰的底物。为了探索SFM4-3的效用，我们现在报告其对2'-叠氮基，2'-氯，2'-氨基或阿拉伯糖的底物识别的表征。我们发现SFM4-3可以有效地合成由这些核苷酸组成的聚合物，最有趣的是，SFM4-3也可以通过PCR扩增这些修饰的寡核苷酸。当与扩增后修饰结合使用时，后者可以对可能以受控方式排列的所需部分进行功能化的聚合物进行指数扩增，

更新日期：2017-10-10

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