Summary
Mutations at the URA3 locus of Saccharomyces cerevisiae can be obtained by a positive selection. Wild-type strains of yeast (or ura3 mutant strains containing a plasmid-borne URA3 + gene) are unable to grow on medium containing the pyrimidine analog 5-fluoro-orotic acid, whereas ura3 − mutants grow normally. This selection, based on the loss of orotidine-5′-phosphate decarboxylase activity seems applicable to a variety of eucaryotic and procaryotic cells.
This is a preview of subscription content, log in via an institution to check access.
References
Bach M, LaCroute F (1972) Direct selective techniques for the isolation of pyrimidine auxotrophs in yeast. Mol Gen Genet 115: 126–130
Bach M, LaCroute F, Botstein D (1979) Evidence for transcriptional regulation of orotidine-5′-phosphate decarboxylase in yeast by hybridization of mRNA to the yeast structural gene cloned in Escherichia coli. Proc Natl Acad Sci USA 76: 386–391
Botstein D, Falco SC, Stewart SE, Brennan M, Scherer S, Stinchomb D, Struhl K, Davis RW (1979) Sterile host yeasts (SHY): A eukaryotic system of biological containment for recombinant DNA materials. Gene 8: 17–24
Chattoo BB, Sherman F, Azubalis DA, Fjellstedt TA, Mehvert D, Ogur M (1979) Selection of lys2 mutants of the yeast Saccharomyces cerevisiae by the utilization of α-amino-adipate. Genetics 93: 51–65
Jund R, LaCroute F (1972) Regulation of orotidylic acid pyrophosphorylase in Saccharomyces cerevisiae. J Bacteriol 109: 196–202
Orr-Weaver TL, Szostak JW, Rothstein RJ (1983) Genetic applications of yeast transformation with linear and gapped plasmids. In: Wu R, Grossman L, Moldave K (eds) Methods in enzymology: Recombinant DNA, Part B. Academic Press, New York, p 228
Rose M, Winston F (1984) Identification of a Ty insertion within the coding sequence of the Saccharomyces cerevisiae URA3 gene. Mol Gen Genet 193: 557–560
Rose M, Grisafi P, Botstein D (1984) Structure and function of the yeast URA3 gene: Expression in E. coli. Gene, in press
Singh A, Sherman F (1974) Characteristics and relationships of mercury-resistant mutants and methionine auxotrophy of yeast. J Bacteriol 118: 911–918
Singh A, Sherman F (1975) Genetic and physiological characterization of met15 mutants of Saccharomyces cerevisiae: A selective system for forward and reverse mutations. Genetics 81: 75–97
Winston F, Chumley FG, Fink GR (1983) Eviction and transplacement of mutant genes in yeast. In: Wu R, Grossman L, Moldave D (eds) Methods in enzymology: Recombinant DNA, Part B. Academic Press, New York, p 211
Winston F, Chaleff DT, Valent B, Fink GR (1984) Mutations affecting Ty-mediated expression of the HIS4 gene of Saccharomyces cerevisiae. Genetics 107: 179–197
Author information
Authors and Affiliations
Additional information
Communicated by H. Saedler
Rights and permissions
About this article
Cite this article
Boeke, J.D., La Croute, F. & Fink, G.R. A positive selection for mutants lacking orotidine-5′-phosphate decarboxylase activity in yeast: 5-fluoro-orotic acid resistance. Mol Gen Genet 197, 345–346 (1984). https://doi.org/10.1007/BF00330984
Received:
Issue Date:
DOI: https://doi.org/10.1007/BF00330984