Development of a translation system using expanded genetic alphabets through unnatural base pairs

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For the expansion of the genetic alphabet of DNA, we studied several codon-anticodon interactions in a cell-free translation system involving hydrophobic unnatural bases. For this project, we mainly worked on to establish efficient transcription systems and their template DNA preparation methods which allow incorporation of hydrophobic unnatural bases at desired potions into DNA and RNA. The results found from this project would open up further development on a translation system involving unnatural base pairs orthogonal to natural base pairs.