Construction of an Opal Suppressor by Oligonucleotide-Directed Mutagenesis of a Saccharomyces cerevisiae tRNA Gene

Authors

Audrey L. Atkin, University of Nebraska-LincolnFollow
K. L. Roy, University of Alberta
J. B. Bell, University of Alberta

Date of this Version

1990

Citation

Molecular and Cellular Biology, Vol. 10, No. 8, Aug. 1990, p. 4379-4383

Comments

Copyright © 1990, American Society for Microbiology

Abstract

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.