Site-specific recombination of yeast 2-micron DNA in vitro.

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Abstract
Most strains of the yeast S. cerevisiae harbor several copies of a 2-.mu.m plasmid circle DNA termed 2 micron. This circular plasmid contains two 599-base-pair precise inverted repeats across which a site-specific conversion event occurs in vivo. This inversion is promoted by a plasmid-encoded function called FLP. The FLP gene of 2-.mu.m DNA under control of a strong yeast promoter was cloned and yeast cells were transformed with a plasmid containing the cloned FLP gene. Cell-free extracts from such a transformant promote highly efficient inversion of 2-.mu.m DNA in vitro. The reaction requires a cation and works efficiently on supercoiled, relaxed circular, or linear DNA. The FLP activity bears certain similarities to the cre protein, a site-specific recombinase encoded by bacteriophage P1.