期刊名称：Proceedings of the National Academy of Sciences
印刷版ISSN：0027-8424
电子版ISSN：1091-6490
出版年度：1980
卷号：77
期号：11
页码：6329-6333
DOI：10.1073/pnas.77.11.6329
语种：English
出版社：The National Academy of Sciences of the United States of America
摘要：A method is presented for isolating DNA segments capable of autonomous replication from Saccharomyces 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. We have cloned from total yeast DNA a number of DNA segments 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 DNAs carried in some of these self-replicating plasmids divides them into two categories: those that are unique in the yeast genome and those that are repetitive.
