Bioorganic & Medicinal Chemistry Letters 2009-07-15

One plasmid selection system for the rapid evolution of aminoacyl-tRNA synthetases.

Charles E Melançon, Peter G Schultz

Index: Bioorg. Med. Chem. Lett. 19 , 3845-3847, (2009)

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Abstract

We have developed a rapid, straightforward, one plasmid dual positive/negative selection system for the evolution of aminoacyl-tRNA synthetases with altered specificities in Escherichia coli. This system utilizes an amber stop codon containing chloramphenicol acetyltransferase/uracil phosphoribosyltransferase fusion gene. We demonstrate the utility of the system by identifying a variant of the Methanococcus jannaschii tyrosyl synthetase from a library of 10(9) variants that selectively incorporates para-iodophenylalanine in response to an amber stop codon.