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Several plasmids, each containing a segment of DNA capable of conferring mitotic stability on the ars1 vector YRp7 replicating in yeast (Saccharomyces cerevisiae), have been isolated from a yeast genomic library by direct selection for stable plasmid maintenance. The mitotic-stabilizing sequences fall into two classes: (i) segments of the yeast "2-micrometers" plasmid and (ii) DNA segments containing yeast centromeric DNA. Plasmids of the first class segregate 4+:0- through meiosis whereas the centromere plasmids segregate predominantly 2+:2-, as typical chromosomes. Previously identified centromeres from chromosomes II (CEN3) and XI (CEN11), along with the CEN11-linked gene MET14, can be isolated by this direct selection procedure. In addition, several other unidentified centromere DNAs were obtained. There are no detectable sequence homologies between CEN3 and CEN11 DNAs nor do these DNAs hybridize to the other yeast centromere regions.

Direct selection procedure for the isolation of functional centromeric DNA.