Patent ID: 7993872

Claim:
A eukaryotic cell comprising an orthogonal aminoacyl-tRNA synthetase (O-RS), wherein the O-RS preferentially aminoacylates an orthogonal tRNA (o-tRNA) with at least one unnatural amino acid in the eukaryotic cell, which unnatural amino acid comprises p-propargyloxyphenylalanine or p-azido-L-phenylalanine, wherein: (a) the O-RS or a portion thereof is encoded by a polynucleotide sequence as set forth in any one of SEQ ID NO: 20-35; (b) the O-RS comprises an amino acid sequence as set forth in any one of SEQ ID NO: 48-63; or, (c) the O-RS comprises an amino acid sequence that is at least 98% identical to that of a naturally occurring tyrosyl aminoacyl-tRNA synthetase (TyrRS) of SEQ ID NO: 2 and comprises two or more amino acids selected from the group consisting of: glycine, serine, or alanine at a position corresponding to Tyr37 of E. coli TyrRS; aspartate at a position corresponding to Asn126 of E. coli TyrRS; asparagine at a position corresponding to Asp182 of E. coli TyrRS; alanine, or valine, at a position corresponding to Phe183 of E. coli TyrRS; and, methionine, valine, cysteine, or threonine, at a position corresponding to Leu186 of E. coli TyrRS.