کد مقاله کد نشریه سال انتشار مقاله انگلیسی نسخه تمام متن
1997863 1065622 2006 8 صفحه PDF دانلود رایگان
عنوان انگلیسی مقاله ISI
Isolating Apparently Pure Libraries of Replication Origins from Complex Genomes
موضوعات مرتبط
علوم زیستی و بیوفناوری بیوشیمی، ژنتیک و زیست شناسی مولکولی زیست شیمی
پیش نمایش صفحه اول مقاله
Isolating Apparently Pure Libraries of Replication Origins from Complex Genomes
چکیده انگلیسی

SummaryBecause 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 ∼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.

ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: - Volume 21, Issue 5, 3 March 2006, Pages 719–726
نویسندگان
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