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Curing ofSaccharomyces cerevisiae2-μm DNA by transformation

✍ Scribed by Elke Erhart; Cornelis P. Hollenberg

Publisher
Springer-Verlag
Year
1981
Tongue
English
Weight
963 KB
Volume
3
Category
Article
ISSN
0172-8083

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✦ Synopsis

A general procedure for the curing of 2-/~m in Saccharomyces cerevisiae is described. The method is based on the displacement of endogenous 2-~m DNA by the recombinant plasmid pMP78-1, which carries the yeast leu2 gene and the 2-/am DNA replicon, but cannot be maintained stably in a yeast cell without endogenous 2-/ma DNA. After transformation with pMP78-1 cells are grown selectively to displace 2-gin DNA. During the non-selective growth which follows, plasmid pMP78-1 is lost and up to 100% of the ceils 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 ° strains has been compared.

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