Summary
A general procedure for the curing of 2-μm in Saccharomyces cerevisiae is described. The method is based on the displacement of endogenous 2-μm DNA by the recombinant plasmid pMP78-1, which carries the yeast leu2 gene and the 2 -μm DNA replicon, but cannot be maintained stably in a yeast cell without endogenous 2-μm DNA. After transformation with pMP78-1 cells are grown selectively to displace 2-μm DNA. During the non-selective growth which follows, plasmid pMP78-1 is lost and up to 100% of the cells completely lack plasmids. In conjunction with a kanamycin resistance marker, as present in plasmid pMP81, this method should be applicable to cure any wild-type yeast strain. The stability of recombinant plasmids in cir + and cir 0 strains has been compared.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Beggs JD (1978) Nature 275:104–109
Blanc H, Gerbaud C, Slonimski PP, Guerineau M (1979) Mol Gen Genet 176:335–342
Bolivar F (1978) Gene 4:121–136
Broach JR, Hicks JB (in press) In: Von Wettstein D, Friis J, Kielland-Brandt M, Stenderup A (eds) Molecular genetics in yeast. Alfred Benzon Symp 16 Munksgaard Copenhagen
Chevallier MR, Aigle M (1979) FEBS Lett 108:179–180
Dobson MJ, Futcher AB, Cox BC (1980) Curr Genet 2:201–205
Gerbaud C, Guerineau M (1980) Curr Genet 1:219–228
Grunstein M, Hogness DS (1975) Proc Natl Acad Sci USA 72:3961–3965
Guerineau M, Grandchamp C, Paoletti J, Slonimski P (1971) Biochem Biophys Res Commun 42:550–557
Hartley JL, Donelson JE (1980) Nature 286:860–865
Hinnen A, Hicks JB, Fink GR (1978) Proc Natl Acad Sci USA 75:1929–1933
Hollenberg CP (1978) Mol Gen Genet 162:23–34
Hollenberg CP (1979) In: Cummings DJ, Borst P, Dawid IB, Weissman SM, Fox CF (eds) Extrachromosomal DNA. ICN-UCLA Symp 15:325–338 Academic Press New York
Hollenberg CP, Borst P, van Bruggen EFJ (1970) Biochim Biophys Acta 209:1–15
Hollenberg CP, Kustermann-Kuhn B, Mackedonski V, Erhart E (in press) In: Von Wettstein D, Friis J, Kielland-Brandt M, Stenderup A (eds) Molecular genetics in yeast. Alfred Benzon Symp 16 Munksgaard Copenhagen
Jimenez A, Davis J (1980) Nature 287:869–871
Livingston DM (1977) Genetics 86:73–84
Maniatis T, Jeffrey A, Kleid DG (1975) Proc Natl Acad Sci USA 72:1184–1188
Saunders GW, Rank GH, Kustermann-Kuhn B, Hollenberg CP (1979) Mol Gen Genet 175:45–52
Southern EM (1975) J Mol Biol 98:503–517
Stewart GG, Russell I, Panchal CJ (1980) Abstracts VI Int Symp on Yeasts London (Ontario), Canada, p 212
Struhl K, Stinchcomb DT, Scherer S, Davis RW (1979) Proc Natl Acad Sci USA 76:1035–1039
Tabak HF (1977) FEBS Letters 84:67–70
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Erhart, E., Hollenberg, C.P. Curing of Saccharomyces cerevisiae 2-μm DNA by transformation. Current Genetics 3, 83–89 (1981). https://doi.org/10.1007/BF00365710
Received:
Issue Date:
DOI: https://doi.org/10.1007/BF00365710