Abstract
We have developed a second orthogonal tRNA/synthetase pair for use in yeast based on the Escherichia coli tRNALeu/leucyl tRNA-synthetase pair. Using a novel genetic selection, we have identified a series of synthetase mutants that selectively charge the amber suppresor tRNA with the C8 amino acid, alpha-aminocaprylic acid, and the photocaged amino acid, o-nitrobenzyl cysteine, allowing them to be inserted into proteins in yeast in response to the amber nonsense codon, TAG.
Publication types
-
Research Support, Non-U.S. Gov't
-
Research Support, U.S. Gov't, P.H.S.
MeSH terms
-
Base Sequence
-
Caprylates / metabolism*
-
Cysteine / analogs & derivatives*
-
Cysteine / metabolism
-
Histidine / genetics
-
Histidine / metabolism
-
Humans
-
Leucine-tRNA Ligase / genetics*
-
Leucine-tRNA Ligase / metabolism*
-
Methyltyrosines / genetics
-
Methyltyrosines / metabolism*
-
Models, Molecular
-
Molecular Sequence Data
-
Mutagenesis
-
Nucleic Acid Conformation
-
Photochemistry
-
Proteins / genetics
-
RNA, Transfer, Amino Acyl / genetics
-
RNA, Transfer, Amino Acyl / metabolism
-
Superoxide Dismutase / genetics
Substances
-
Caprylates
-
Methyltyrosines
-
Proteins
-
RNA, Transfer, Amino Acyl
-
O-methyltyrosine
-
Histidine
-
Superoxide Dismutase
-
Leucine-tRNA Ligase
-
Cysteine