Expression of a set of synthetic suppressor tRNA(Phe) genes in Saccharomyces cerevisiae.
Synthetic ochre and amber tRNA suppressor genes derived from the yeast tRNA(PheGAA) sequence have been constructed. They were efficiently transcribed in vitro and expressed in vivo via a synthetic expression cassette. tRNA(PheUUA) and tRNA(PheUUA) delta IVS (IVS = intervening sequence) are relatively inefficient ochre suppressors. They are toxic to the cell when expressed on a multicopy plasmid, and they do not suppress at all when present as single copies. The intron does not seem to have any effect on suppression. In contrast, the amber suppressor tRNA(PheCUA) delta IVS is efficient when expressed from a single-copy plasmid, while its efficiency is reduced on a multicopy vector.
Masson
J M
JM
Department of Biology, University of California, Los Angeles 90024.
Meuris
P
P
Grunstein
M
M
Abelson
J
J
Miller
J H
JH
D001483:Base Sequence; D004926:Escherichia coli / Q000235:genetics; D005800:Genes, Fungal; D005813:Genes, Synthetic; D008969:Molecular Sequence Data; D009690:Nucleic Acid Conformation; D010957:Plasmids; D012345:RNA, Transfer, Amino Acid-Specific / Q000235:genetics*; D012360:RNA, Transfer, Phe / Q000235:genetics*; D012441:Saccharomyces cerevisiae / Q000235:genetics*; D013489:Suppression, Genetic; D014158:Transcription, Genetic
D012345:RNA, Transfer, Amino Acid-Specific; D012360:RNA, Transfer, Phe