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Gene Synthesis> | … 2.2. In vitro transcription. The tRNA Leu (UAG) gene was synthesized and subsequently cloned into the M13mp8 [25] vector by GenScript (Piscataway, NJ) to create a plasmid construct similar to the plasmid created by Normanly et al. [25] … | Get A Quote |
RNA is known to interact with Mg when assuming higher-ordered tertiary configurations. Structurally, when tRNA molecules interact with Mg, they consistently form a "L-shape" conformation each time they are synthesized. Therefore, if Mg can induce tertiary structure formation, then binding to alternative cations could produce alternative tertiary configurations. By utilizing circular dichroism and mobility gel-shift assays it was observed that tRNA structure can be altered when in the presence of different divalent cationic species. Formation of these alternative structural configurations was further validated by aminoacylating these tRNA structural anomalies with their native enzyme, which resulted in... More