One plasmid selection system for the rapid evolution of aminoacyl-tRNA synthetases
Charles E Melançon1, Peter G Schultz
1Department of Chemistry, The Scripps Research Institute, La Jolla, CA 92037, USA.
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.
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