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Vol. 12, Issue 11, 3317-3327, November 2001
and
Department of Microbiology and Molecular Genetics, UMDNJ-New Jersey
Medical School, Newark, New Jersey 07103
In Saccharomyces cerevisiae chromosomal DNA
replication initiates at intervals of ~40 kb and depends upon the
activity of autonomously replicating sequence (ARS) elements.
The identification of ARS elements and analysis of their function as
chromosomal replication origins requires the use of functional assays
because they are not sufficiently similar to identify by DNA sequence analysis. To complete the systematic identification of ARS elements on
S. cerevisiae chromosome III, overlapping clones
covering 140 kb of the right arm were tested for their ability to
promote extrachromosomal maintenance of plasmids. Examination of
chromosomal replication intermediates of each of the seven ARS elements
identified revealed that their efficiencies of use as chromosomal
replication origins varied widely, with four ARS elements active in
10% of cells in the population and two ARS elements active in
90%
of the population. Together with our previous analysis of a 200-kb
region of chromosome III, these data provide the first complete
analysis of ARS elements and DNA replication origins on an entire
eukaryotic chromosome.
Present address: BD Biosciences, San Jose, CA 95131.
Corresponding author: E-mail address:
newlon{at}umdnj.edu.
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