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Updated: Aug 10, 2026

The Green Monster Process for the Generation of Yeast Strains Carrying Multiple Gene Deletions
Published on: December 15, 2012
Targeting, disruption, replacement, and allele rescue: integrative DNA transformation in yeast
Abstract:
The methods described in this chapter permit the manipulation of virtually any cloned yeast chromosomal sequence by virtue of the fact that DNA transformed into yeast integrates into the chromosome by homologous recombination. Furthermore, double-strand breaks in transforming DNA stimulate recombination and can be used to target integration events. This allows simple one-step gene disruption methods using yeast selectable markers. The availability of counterselectable markers makes it possible to replace chromosomal sequences with mutant alleles that cannot be directly selected. Finally, these same methods can be used to rescue chromosomal alleles on plasmids for subsequent molecular analysis.
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