Introduction of the plasmid pKM101-associated muc genes into Saccharomyces cerevisiae.
Bacteria-yeast shuttle plasmids containing the pKM101-associated muc genes were constructed by cloning an ARS TRP fragment into the plasmid pGW270 in both possible orientations. The insertion of Saccharomyces cerevisiae DNA into pGW270 had no effect on the mutator and protective phenotypes associated with the plasmid in Escherichia coli. Two such recombinant plasmids, pAA90 and pAA91 , were capable of efficient transformation of S. cerevisiae and were stably maintained in this organism. Hybridization experiments suggest that muc-specific mRNA was present in transformed yeast cells and a small amount was polyadenylated. The RNAs were not of a discrete size, all being smaller than the muc genes. The presence of the plasmid pAA91 , and to a lesser extent, pAA90 , in yeast resulted in a detectable increase in the reversion frequencies of three markers and in ultraviolet protection. These results are discussed in terms of studying the relationship of error-prone repair in bacteria and yeast and of developing improved yeast tester strains.
D004260:DNA Repair; D004274:DNA, Recombinant; D005818:Genetic Engineering; D009152:Mutagenicity Tests / Q000379:methods*; D009154:Mutation / Q000528:radiation effects; D010957:Plasmids; D012441:Saccharomyces cerevisiae / Q000235:genetics*; D014466:Ultraviolet Rays
D004274:DNA, Recombinant