부산대학교 도서관

To link to full-text access for this article, visit this link: http://dx.doi.org/10.1016/j.molcel.2006.01.015 Byline: Larry D. Mesner (1), Emily L. Crawford (1), Joyce L. Hamlin (1) Abstract: Because of the complexity of higher eukaryotic genomes and the lack of a reliable autonomously replicating sequence (ARS) assay for isolating potential replicators, the identification of origins has proven to be extremely challenging and time consuming. We have developed a new origin-trapping method based on the partially circular nature of restriction fragments containing replication bubbles and have prepared a library of [approximately equal to]1000 clones from early S phase CHO cells. When 15 randomly selected clones were analyzed by a stringent two-dimensional (2D) gel replicon mapping method, all were shown to correspond to active, early firing origins. Furthermore, most of these appear to derive from broad zones of potential sites, and the five that were analyzed in a time-course study are all inefficient. This bubble-trapping scheme will allow the construction of comprehensive origin libraries from any complex genome so that their natures and distributions vis-a-vis other chromosomal markers can be established. Author Affiliation: (1) Department of Biochemistry and Molecular Genetics, University of Virginia School of Medicine, Charlottesville, Virginia 22908 Article History: Received 24 October 2005; Revised 16 December 2005; Accepted 12 January 2006 Article Note: (miscellaneous) Published: March 2, 2006