Efficient and sequence-independent replication of DNA containing a third base pair establishes a functional six-letter genetic alphabet

Malyshev D.A.; Dhami K.; Quach H.T.Lavergne T.Ordoukhanian P.Torkamani A.Romesberg F.E.

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Efficient and sequence-independent replication of DNA containing a third base pair establishes a functional six-letter genetic alphabet

机译：Efficient and sequence-independent replication of DNA containing a third base pair establishes a functional six-letter genetic alphabet

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摘要

The natural four-letter genetic alphabet, comprised of just two base pairs (dA-dT and dG-dC), is conserved throughout all life, and its expansion by the development of a third, unnatural base pair has emerged as a central goal of chemical and synthetic biology. We recently developed a class of candidate unnatural base pairs, exemplified by the pair formed between d5SICS and dNaM. Here, we examine the PCR amplification of DNA containing one or more d5SICS-dNaM pairs in a wide variety of sequence contexts. Under standard conditions, we show that this DNA may be amplified with high efficiency and greater than 99.9 fidelity. To more rigorously explore potential sequence effects, we used deep sequencing to characterize a library of templates containing the unnatural base pair as a function of amplification. We found that the unnatural base pair is efficiently replicated with high fidelity in virtually all sequence contexts. The results show that, for PCR and PCR-based applications, d5SICS-dNaM is functionally equivalent to a natural base pair, and when combined with dA-dT and dG-dC, it provides a fully functional six-letter genetic alphabet.

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《Proceedings of the National Academy of Sciences of the United States of America》 |2012年第30期|12005-12010|共6页
作者
Malyshev D.A.; Dhami K.; Quach H.T.Lavergne T.Ordoukhanian P.Torkamani A.Romesberg F.E.;
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作者单位

Department of Chemistry, Scripps Research Institute, San Diego, CA 92037, United States;

Center for Protein and Nucleic Acid Research, Scripps Research Institute, San Diego, CA 92037, United States;

Scripps Translational Science Institute, Scripps Research Institute, San Diego, CA 92037, United States;

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收录信息美国《科学引文索引》(SCI);美国《生物学医学文摘》(MEDLINE);美国《化学文摘》(CA);
原文格式 PDF
正文语种英语
中图分类自然科学总论;
关键词
Artificial DNA; Bioinformatics; Expanded genetic alphabet; Hydrophobic; Unnatural nucleotides;

Efficient and sequence-independent replication of DNA containing a third base pair establishes a functional six-letter genetic alphabet

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