纳米孔测序扩展的遗传密码揭示了主要疏水性非天然碱基对的高保真复制。
Nanopore Sequencing of an Expanded Genetic Alphabet Reveals High-Fidelity Replication of a Predominantly Hydrophobic Unnatural Base Pair.
机构信息
Department of Chemistry , The Scripps Research Institute , La Jolla , California 92037 , United States.
Department of Physics , University of Washington , Seattle , Washington 98195 , United States.
出版信息
J Am Chem Soc. 2020 Feb 5;142(5):2110-2114. doi: 10.1021/jacs.9b09808. Epub 2020 Jan 27.
Abstract
Unnatural base pairs (UBPs) have been developed and used for a variety of applications as well as for the engineering of semisynthetic organisms (SSOs) that store and retrieve increased information. However, these applications are limited by the availability of methods to rapidly and accurately determine the sequence of unnatural DNA. Here we report the development and application of the MspA nanopore to sequence DNA containing the d-d UBP. Analysis of two sequence contexts reveals that DNA containing the UBP is replicated with an efficiency and fidelity similar to that of natural DNA and sufficient for use as the basis of an SSO that produces proteins with noncanonical amino acids.
摘要
非天然碱基对（UBP）已经被开发并应用于各种领域，包括用于存储和检索更多信息的半合成生物体（SSO）的工程改造。然而，这些应用受到了缺乏快速、准确确定非天然 DNA 序列方法的限制。在这里，我们报告了 MspA 纳米孔的开发和应用，用于对含有 d-d UBP 的 DNA 进行测序。对两种序列结构的分析表明，含有 UBP 的 DNA 的复制效率和保真度与天然 DNA 相似，足以作为产生具有非规范氨基酸的蛋白质的 SSO 的基础。
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