Site-Specific Recombination Promoted in Vitro by the FLP Protein of the Yeast Two-Micron Plasmid
The yeast 2-micron plasmid is a 6,318-bp circular DNA molecule occurring in many yeast strains. It is generally present at 50 to 100 copies per cell and has been sequenced in its entirety (11). The plasmid encodes at least four protein products (Ref. 6; Volkert et al., this Volume), which are involved primarily in plasmid maintenance. One of these proteins, designated FLP, is the central component in a plasmid-encoded, site-specific recombination system. The sequence of the open reading frame (ORF) encoding this protein provides a predicted molecular weight of slightly less than 48,000 Da. This recombinase acts at a pair of sites located within the 599-bp inverted repeats. Recombination results in the inversion of the DNA between the sites, giving two forms of the plasmid, termed A and B (see Fig. 1). Isolated plasmid DNA is found to be a 50:50 mixture of the two forms. Inversion has been shown to be part of a copy-number amplification system (Volkert et al., this Volume). The general features of the plasmid and its maintenance systems appear to be shared by plasmids isolated from a variety of related organisms (A. Toh-e et al., this Volume).
KeywordsRecombination Site XbaI Restriction Site Yeast Plasmid Homology Requirement Phosphate Contact
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