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Tools to generate an orthogonal LeuRSaRNA Leu pair were developed by using a two plasmid system in an E. coli strain containing a nonfunctional genomic lacZ gene with an amber stop codon. The plasmid pRAH curies tRNALeu with an amber suppressor anticodon, while pKEG harbors LeuRS. From this system, directed evolution approaches can be used to screen for an orthogonal tRNA and a LeuRS derivative that will charge it.","Made available in DSpace on 2015-09-25T22:28:14Z (GMT). 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