An improved method for yeast 2 microns plasmid curing.
SMR1-410, a dominant resistance marker, was cloned into the FLP gene of 2 microns DNA to produce the chimeric YEp vector pWX823B. Selection for SMR1-410 at high concentrations of sulfometuron methyl maintained pWX823 at high copy number and resulted in the rapid and efficient loss of native 2 microns DNA. Using this protocol approximately 15% of the cells monitored showed loss of 2 microns DNA. The curing methodology is more efficient and convenient than previous methods and has the added advantage of being applicable to wild-type prototrophic cells.
Department of Biology, University of Saskatchewan, Saskatoon, Canada.
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D003001:Cloning, Molecular / Q000379:methods*; D004352:Drug Resistance, Microbial / Q000235:genetics; D005819:Genetic Markers; D010957:Plasmids; D012441:Saccharomyces cerevisiae / Q000235:genetics*; D013453:Sulfonylurea Compounds / Q000494:pharmacology
D005819:Genetic Markers; D013453:Sulfonylurea Compounds; C042031:sulfometuron methyl