Nature Methods 3, 263 - 265 (2006)
Published online: 22 March 2006; | doi:10.1038/nmeth864
Efficient incorporation of unnatural amino acids into proteins in Escherichia coliYoungha Ryu
& Peter G Schultz
Department of Chemistry and the Skaggs Institute for Chemical Biology, The Scripps Research Institute, 10550 North Torrey Pines Road, La Jolla, California 92037, USA.
Correspondence should be addressed to Peter G Schultz schultz@scripps.edu We have developed a single-plasmid system for the efficient bacterial expression of mutant proteins containing unnatural amino acids at specific sites designated by amber nonsense codons. In this system, multiple copies of a gene encoding an amber suppressor tRNA derived from a Methanocaldococcus jannaschii tyrosyl-tRNA (MjtRNATyrCUA) are expressed under control of the proK promoter and terminator, and a gene encoding the desired mutant M. jannaschii tyrosyl-tRNA synthetase (MjTyrRS) is expressed under control of a mutant glnS (glnS') promoter.
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