An unnatural hydrophobic base pair system: site-specific incorporation of nucleotide analogs into DNA and RNA
作者： Ichiro Hirao , Michiko Kimoto , Tsuneo Mitsui , Tsuyoshi Fujiwara , Rie Kawai
DOI: 10.1038/NMETH915
关键词: Polymerase 、 RNA 、 Base pair 、 Nucleic acid secondary structure 、 Stereochemistry 、 Transcription (biology) 、 T7 RNA polymerase 、 Wobble base pair 、 Biochemistry 、 Biology 、 DNA
摘要: Methods for the site-specific incorporation of extra components into nucleic acids can be powerful tools creating DNA and RNA molecules with increased functionality. We present an unnatural base pair system in which containing amplified function as a template analog substrates via transcription. The is formed by specific hydrophobic shape complementation between bases, but lacks hydrogen bonding interactions. In replication, this exhibits high selectivity combination usual triphosphates modified triphosphates, gamma-amidotriphosphates, 3' to 5' exonuclease-proficient polymerases, allowing PCR amplification. transcription, complementarity mediates these their analogs, such biotinylated substrate, T7 polymerase (RNAP). With system, functional site-specifically incorporated large molecule.
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