title: Transforming a pair of orthogonal tRNA-aminoacyl-tRNA synthetase from Archaea to function in mammalian cells.
abstract: A previously engineered Methanocaldococcus jannaschii tRNA(CUA Tyr)-tyrosyl-tRNA synthetase pair orthogonal to Escherichia coli was modified to become orthogonal in mammalian cells. The resulting tRNA(CUA Tyr)-tyrosyl-tRNA synthetase pair was able to suppress an amber codon in the green fluorescent protein, GFP, and in a foldon protein in mammalian cells. The methodology reported here will allow rapid transformation of the much larger collection of existing tyrosyl-tRNA synthetases that were already evolved for the incorporation of an array of over 50 unnatural amino acids into proteins in Escherichia coli into proteins in mammalian cells. Thus we will be able to introduce a large array of possibilities for protein modifications in mammalian cells.
authors.affiliation: Division of Medicinal Chemistry, College of Pharmacy, University of Texas at Austin, Austin, Texas, United States of America.
authors.forename: Gabrielle Nina
authors.initials: GN
authors.lastname: Thibodeaux
authors.forename: Xiang
authors.initials: X
authors.lastname: Liang
authors.forename: Kathryn
authors.initials: K
authors.lastname: Moncivais
authors.forename: Aiko
authors.initials: A
authors.lastname: Umeda
authors.forename: Oded
authors.initials: O
authors.lastname: Singer
authors.forename: Lital
authors.initials: L
authors.lastname: Alfonta
authors.forename: Zhiwen Jonathan
authors.initials: ZJ
authors.lastname: Zhang
mesh_terms: D000604:Amino Acyl-tRNA Synthetases / Q000235:genetics / Q000378:metabolism*; D000818:Animals; D001105:Archaea / Q000201:enzymology*; D001483:Base Sequence; D015153:Blotting, Western; D002460:Cell Line; D006801:Humans; D008322:Mammals; D008969:Molecular Sequence Data; D009690:Nucleic Acid Conformation; D012343:RNA, Transfer / Q000737:chemistry / Q000235:genetics*
chemical_list: D012343:RNA, Transfer; D000604:Amino Acyl-tRNA Synthetases