AI Article Synopsis
- The development of a new unnatural base pair system (Ds-Pn) enhances genetic encoding and offers significant potential for biotechnology applications.
- Unlike previous unnatural pairs, this system shows improved selectivity during DNA replication, effectively minimizing mispairing with natural bases.
- The newly engineered pair exhibits a mutation rate of about 1% after PCR amplification, suggesting its practicality for expanding genetic systems in various biotechnological applications.
Article Abstract
Expansion of the genetic alphabet by an unnatural base pair system provides a powerful tool for modern biotechnology. As an alternative to previous unnatural base pairs, we have developed a new pair between 7-(2-thienyl)imidazo[4,5-b]pyridine (Ds) and 2-nitropyrrole (Pn), which functions in DNA amplification. Pn more selectively pairs with Ds in replication than another previously reported pairing partner, pyrrole-2-carbaldehyde (Pa). The nitro group of Pn efficiently prevented the mispairing with A. High efficiency and selectivity of the Ds-Pn pair in PCR amplification were achieved by using a substrate mixture of the gamma-amidotriphosphate of Ds and the usual triphosphates of Pn and the natural bases, with Vent DNA polymerase as a 3' to 5' exonuclease-proficient polymerase. After 20 cycles of PCR, the total mutation rate of the Ds-Pn site in an amplified DNA fragment was approximately 1%. PCR amplification of DNA fragments containing the unnatural Ds-Pn pair would be useful for expanded genetic systems in DNA-based biotechnology.
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| http://dx.doi.org/10.1021/ja073830m | DOI Listing |
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