Efficient and sequence-independent replication of DNA containing a third base pair establishes a functional six-letter genetic alphabet
| Autor: | Ali Torkamani, Thomas Lavergne, Henry T. Quach, Kirandeep Dhami, Floyd E. Romesberg, Denis A. Malyshev, Phillip Ordoukhanian |
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| Rok vydání: | 2012 |
| Předmět: |
DNA Replication
Base pair Molecular Sequence Data Oligonucleotides Electrophoretic Mobility Shift Assay Computational biology Biology Polymerase Chain Reaction Deep sequencing Synthetic biology chemistry.chemical_compound Sequence (medicine) Genetics Multidisciplinary Base Sequence Molecular Structure Oligonucleotide Nucleotides DNA replication dNaM Computational Biology High-Throughput Nucleotide Sequencing Biological Sciences chemistry Synthetic Biology Genetic Engineering DNA |
| Zdroj: | Proceedings of the National Academy of Sciences of the United States of America. 109(30) |
| ISSN: | 1091-6490 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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