Summary
A new method for the routine isolation of mutations of spontaneous origin in the yeast, Saccharomyces cerevisiae, is reported. The technique is based on the observation that inositol auxotrophs die when deprived of inositol. However, if marromolecular synthesis is inhibited, most of the cells survive. Appropriate manipulation of inositol requiring mutants can threfore result in the selective survival of cells possesing mutations which affect macromolecular synthesis. Since reversion to inositol prototrophy can be a major source of interference in efficient selection, a haploid double mutant strain has been constructed which reverts to inositol prototrophy with a frequency estimated to be several orders of magnitude lower than the expected frequency of single, spontaneous mutational events. Using this strain, enrichment in excess of 10000 fold has been obtained for various classes of auxotrophic mutants. Spontaneous temperature sensitive mutants have also been obtained.
Similar content being viewed by others
References
Culbertson, M.R., Henry, S.A.: Inositol requiring mutants of Saccharomyces cerevisiae. Genetics 80, 23–40 (1975)
Ferenczy, L., Sipiczki, M., Szegedi, M.: Enrichment of fungal mutants by selective cell wall lysis. Nature (Lond.) 253, 46–47 (1975)
Fink, G.R., Styles, C.A.: Gene conversion of deletions in the his 4 region of yeast. Genetics 77, 231–244 (1974)
Fogel, S., Hurst, D.D.: Meiotic gene conversion in yeast tetrads and the theory of recombination. Genetics 57, 455–481 (1967)
Henry, S.A., Horowitz, B.: A new method for mutant selection in Saccharomyces cerevisiae. Genetics 79, 175–186 (1975)
Holliday, R.: Selection of auxotrophs by inositiol starvation in Ustilago maydis. Microbial. Gen. Bull. Mo. 13, 28–30 (1962)
Lester, H.E., Gross, S.R.: Efficient method of selection of auxotrophic mutants of Neurospora. Science 139, 572 (1959)
Lindegren, G.. Hwang, Y.L., Oshima, Lindegren, C.C.: Genetical mutants induced by ethyl methanesulfonate in Saccharomyces. Canad. Genet. Cytol. 7, 491–499 (1965)
Littlewood, B.S.: A method of obtaining antibiotic sensitive mutants in Saccharomyces cerevisiae. Genetics 71, 305–308 (1972)
Littlewood, B.S., Davies, J.E.: Enrichment for temperature sensitive and auxotrophic mutants in Saccharomyces cerevisiae by tritium suicide. Mutation Res. 17, 315–322 (1973)
Luria, S.E., Delbrück, M.: Mutations of bacteria from virus sensitivity to virous resistance. Genetics 28, 491–511 (1943)
Snow, R.: An enrichment method of auxotrophic yeast mutants using the antibiotic “Nystatin”. Nature (Lond.) 211, 206–207 (1966)
Thomas, P.L.: Increased frequency of auxotrophic mutants of Ustilago hordei affer combined UV irradiation and inositol starvation. Canad. J. Genet. Cytol. 14, 785–788 (1972)
Author information
Authors and Affiliations
Additional information
Communicated by F. Stahl
Rights and permissions
About this article
Cite this article
Henry, S.A., Donahue, T.F. & Culbertson, M.R. Selection of spontaneous mutants by inositol starvation in yeast. Molec. Gen. Genet. 143, 5–11 (1975). https://doi.org/10.1007/BF00269415
Received:
Issue Date:
DOI: https://doi.org/10.1007/BF00269415