Autonomously replicating sequences in Saccharomyces cerevisiae.

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Abstract
A method is presented for isolating DNA segments capable of autonomous replication from S. cerevisiae chromosomal DNA based on the differential transforming ability of autonomously replicating plasmids and nonreplicating plasmids. DNA plasmids that are capable of self-replication in yeast transform yeast spheroplasts at about 1000-fold higher frequency than nonreplicating plasmids. From total yeast DNA a number of DNA segments were cloned that permit the pBR322 plasmid carrying the yeast LEU2 gene to replicate autonomously. These plasmid clones are characterized by their ability to transform Leu- spheroplasts to Leu+ at a high frequency and their ability to replicate autonomously. Analysis of the insert DNA carried in some of these self-replicating plasmids divides them into 2 categories: those that are unique in the yeast genome and those that are repetitive.