ORTHOGONAL TRANSLATION COMPONENTS FOR THE IN VIVO INCORPORATION OF UNNATURAL AMINO ACIDS

Abstract

The invention relates to orthogonal pairs of tRNAs and cognate aminoacyl-tRNA synthetases that can incorporate unnatural amino acids into proteins produced in eubacterial host cells such as E. coli, or in a eukaryotic host such as a yeast cell. The invention provides novel orthogonal synthetases, methods for identifying and making the novel synthetases, methods for producing proteins contg. unnatural amino acids, and translation systems. In particular, the amino acid sequences and the encoding nucleotide sequences of mutant Methanococcus jannaschii tyrosyl-tRNA synthetase and Escherichia coli leucyl-tRNA synthetase are disclosed. The synthetases were modified to accept a range of amino acid analogs as substrates for the charging of the tRNA. The tRNA were also modified to create a four- or five base anticodon that can be used to limit the incorporation of the foreign amino acid to specific sites. [on SciFinder(R)]