Premium
Mapping replication origins in yeast chromosomes
Author(s) -
Brewer Bonita J.,
Fangman Walton L.
Publication year - 1991
Publication title -
bioessays
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.175
H-Index - 184
eISSN - 1521-1878
pISSN - 0265-9247
DOI - 10.1002/bies.950130702
Subject(s) - replicon , biology , origin of replication , genetics , eukaryotic dna replication , dna replication , saccharomyces cerevisiae , context (archaeology) , origin recognition complex , yeast , dna , replication (statistics) , eukaryotic chromosome fine structure , genome , replication timing , chromosome , computational biology , gene , paleontology , virology
The replicon hypothesis, first proposed in 1963 by Jacob and Brenner (1) , states that DNA replication is controlled at sites called origins. Replication origins have been well studied in prokaryotes. However, the study of eukaryotic chromosomal origins has lagged behind, because until recently there has been no method for reliably determining the identity and location of origins from eukaryotic chromosomes. Here, we review a technique we developed with the yeast Saccharomyces cerevisiae that allows both the mapping of replication origins and an assessment of their activity. Two‐dimensional agarose gel electrophoresis and Southern hybridization with total genomic DNA are used to determine whether a particular restriction fragment acquires the branched structure diagnostic of replication initiation. The technique has been used to localize origins in yeast chromosomes and assess their initiation efficiency. In some cases, origin activation is dependent upon the surrounding context. The technique is also being applied to a variety of eukaryotic organisms.