Efficient and sequence-independent replication of DNA containing a third base pair establishes a functional six-letter genetic alphabet
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.
Références
- Titre
- Efficient and sequence-independent replication of DNA containing a third base pair establishes a functional six-letter genetic alphabet
- Type de publication
- Article de revue
- Année de publication
- 2012
- Auteurs
- Malyshev, D. A., Dhami K., Quach H. T., Lavergne Thomas, Ordoukhanian P., Torkamani A., and Romesberg F. E.
- Revue
- Proc Natl Acad Sci U S A
- Volume
- 109
- Pagination
- 12005-10
Soumis le 5 juillet 2018