One plasmid selection system for the rapid evolution of aminoacyl-tRNA synthetases
Dual positive/negative selection vectors which allow the facile evolution of aminoacyl-tRNA synthetases are reported. We have developed a rapid, straightforward, one plasmid dual positive/negative selection system for the evolution of aminoacyl-tRNA synthetases with altered specificities in Escheric...
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Veröffentlicht in: | Bioorganic & medicinal chemistry letters 2009-07, Vol.19 (14), p.3845-3847 |
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Format: | Artikel |
Sprache: | eng |
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Zusammenfassung: | Dual positive/negative selection vectors which allow the facile evolution of aminoacyl-tRNA synthetases are reported.
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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ISSN: | 0960-894X 1464-3405 |
DOI: | 10.1016/j.bmcl.2009.04.007 |