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The in vivo replication origin of the yeast 2μm plasmid

1987; Cell Press; Volume: 51; Issue: 3 Linguagem: Inglês

10.1016/0092-8674(87)90643-x

1097-4172

Joel A. Huberman, Loretta D. Spotila, Kevin A. Nawotka, Sufian M. ElAssouli, Leslie R. Davis,

Genomics and Chromatin Dynamics

Abstract We have used two-dimensional neutral/alkaline agarose gel electrophoresis to separate the nascent strands of replicating yeast 2μm plasmid DNA molecules according to extent of replication, away from nonreplicating molecules and parental strands. Analysis of the lengths of nascent strands by sequential hybridization with short probes shows that replication proceeds bidirectionally from a single origin at map position 3700 ± 100, coincident with the genetically mapped ARS element. The two recombinational isomers of 2μm plasmid (forms A and B) replicate with equal efficiency. These results suggest that ARS elements may prove to be replication origins for chromosomal DNA.