Construction of an opal suppressor by oligonucleotide-directed mutagenesis of a Saccharomyces cerevisiae tRNA(Trp) gene.
In vitro mutagenesis was used to create putative opal suppressor alleles of a tRNA(Trp) gene of Saccharomyces cerevisiae. The construct with the requisite anticodon change did not result in an active suppressor, whereas when a second change was introduced into the portion of the gene encoding the intron, an active and specific opal suppressor was produced. We propose that the secondary structure of transcripts from the first mutant may prevent efficient pre-tRNA processing, whereas normal processing occurs with the double mutant.
Atkin
A L
AL
Department of Genetics, University of Alberta, Edmonton, Canada.
Roy
K L
KL
Bell
J B
JB
D000483:Alleles; D001483:Base Sequence; D003001:Cloning, Molecular; D004271:DNA, Fungal / Q000235:genetics; D005800:Genes, Fungal; D008969:Molecular Sequence Data; D009154:Mutation; D009690:Nucleic Acid Conformation; D015345:Oligonucleotide Probes; D010957:Plasmids; D012345:RNA, Transfer, Amino Acid-Specific / Q000235:genetics*; D012364:RNA, Transfer, Trp / Q000235:genetics*; D015183:Restriction Mapping; D013489:Suppression, Genetic; D014158:Transcription, Genetic
D004271:DNA, Fungal; D015345:Oligonucleotide Probes; D012345:RNA, Transfer, Amino Acid-Specific; D012364:RNA, Transfer, Trp