Natural-like Replication of an Unnatural Base Pair for the Expansion of the Genetic Alphabet and Biotechnology Applications

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• 作者：Lingjun Li ; Mélissa Degardin ; Thomas Lavergne ; Denis A. Malyshev ; Kirandeep Dhami ; Phillip Ordoukhanian ; Floyd E. Romesberg
• 刊名：Journal of the American Chemical Society
• 出版年：2014
• 出版时间：January 22, 2014
• 年：2014
• 卷：136
• 期：3
• 页码：826-829
• 全文大小：235K
• ISSN：1520-5126

文摘

We synthesized a panel of unnatural base pairs whose pairing depends on hydrophobic and packing forces and identify dTPT3-dNaM, which is PCR amplified with a natural base pair-like efficiency and fidelity. In addition, the dTPT3 scaffold is uniquely tolerant of attaching a propargyl amine linker, resulting in the dTPT3^PA-dNaM pair, which is amplified only slightly less well. The identification of dTPT3 represents significant progress toward developing an unnatural base pair for the in vivo expansion of an organism鈥檚 genetic alphabet and for a variety of in vitro biotechnology applications where it is used to site-specifically label amplified DNA, and it also demonstrates for the first time that hydrophobic and packing forces are sufficient to mediate natural-like replication.