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. 1983 Dec;80(23):7284-8.
doi: 10.1073/pnas.80.23.7284.

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

D Vetter, B J Andrews, L Roberts-Beatty, P D Sadowski

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

D Vetter et al. Proc Natl Acad Sci U S A. 1983 Dec.

Abstract

Most strains of the yeast Saccharomyces cerevisiae harbor several copies of a 2-micron plasmid circle DNA termed "2 micron." This circular plasmid contains two 599-base-pair precise inverted repeats across which a site-specific inversion event occurs in vivo. This inversion is promoted by a plasmid-encoded function called "FLP." We have cloned the FLP gene of 2-micron DNA under control of a strong yeast promoter and transformed yeast cells with a plasmid containing the cloned FLP gene. Cell-free extracts from such a transformant promote highly efficient inversion of 2-micron 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.

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