Molecular Dissection of an Extrachromosomal Amplicon Reveals a Circular Structure Consisting of an Imperfect Inverted Duplication
Identifieur interne : 004639 ( Main/Exploration ); précédent : 004638; suivant : 004640Molecular Dissection of an Extrachromosomal Amplicon Reveals a Circular Structure Consisting of an Imperfect Inverted Duplication
Auteurs : Genevieve H. Nonet [États-Unis] ; Susan M. Carroll [États-Unis] ; Margaret L. Derose [États-Unis] ; Geoffrey M. Wahl [États-Unis]Source :
- Genomics [ 0888-7543 ] ; 1993.
Abstract
Abstract: A mouse fibroblast line, B-1/50, with a 4300-fold amplification of the adenosine deaminase gene locus (Yeung et al., 1983, J. Biol. Chem. 258: 8338-8345), was shown by in situ hybridization to harbor the amplified sequences on variously sized extrachromosomal elements. We show here that the smallest circle is approximately 500 kb. We describe a facile screening technique for identifying cosmid and yeast artificial chromosome (YAC) clones derived from the amplicon. A closed molecular map was generated by arranging the cosmids and YACs into a contig spanning over 250 kb of the adenosine deaminase gene locus. YACs from the two ends of this contig were shown to delimit a 250-kb inverted duplication. Long-range mapping of a Sal I partial digest of B-1/50 DNA is also consistent with the interpretation that the 500-kb adenosine deaminase amplicon in B-1/50 cells is an inverted duplication. The finding that this amplicon is the only or predominant structure containing amplified sequences in the B-1/50 cell line suggests that such structures are not inherently prone to high frequency rearrangement, even when present at such high copy number. This study provides the first molecular description of the structure of an episome involved in mammalian gene amplification. The implications of this finding for models of gene amplification and episome formation are discussed.
Url:
DOI: 10.1006/geno.1993.1107
Affiliations:
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Nonet</name><affiliation wicri:level="2"><country xml:lang="fr">États-Unis</country><placeName><region type="state">Californie</region></placeName><wicri:cityArea>Gene Expression Laboratory, The Salk Institute for Biological Studies, 10010 North Torrey Pines Road, La Jolla, California 92037; and Department of Chemistry, University of California at San Diego, La Jolla</wicri:cityArea></affiliation></author><author><name sortKey="Carroll, Susan M" sort="Carroll, Susan M" uniqKey="Carroll S" first="Susan M." last="Carroll">Susan M. 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Biol. Chem. 258: 8338-8345), was shown by in situ hybridization to harbor the amplified sequences on variously sized extrachromosomal elements. We show here that the smallest circle is approximately 500 kb. We describe a facile screening technique for identifying cosmid and yeast artificial chromosome (YAC) clones derived from the amplicon. A closed molecular map was generated by arranging the cosmids and YACs into a contig spanning over 250 kb of the adenosine deaminase gene locus. YACs from the two ends of this contig were shown to delimit a 250-kb inverted duplication. Long-range mapping of a Sal I partial digest of B-1/50 DNA is also consistent with the interpretation that the 500-kb adenosine deaminase amplicon in B-1/50 cells is an inverted duplication. The finding that this amplicon is the only or predominant structure containing amplified sequences in the B-1/50 cell line suggests that such structures are not inherently prone to high frequency rearrangement, even when present at such high copy number. This study provides the first molecular description of the structure of an episome involved in mammalian gene amplification. The implications of this finding for models of gene amplification and episome formation are discussed.</div></front></TEI><affiliations><list><country><li>États-Unis</li></country><region><li>Californie</li></region></list><tree><country name="États-Unis"><region name="Californie"><name sortKey="Nonet, Genevieve H" sort="Nonet, Genevieve H" uniqKey="Nonet G" first="Genevieve H." last="Nonet">Genevieve H. Nonet</name></region><name sortKey="Carroll, Susan M" sort="Carroll, Susan M" uniqKey="Carroll S" first="Susan M." last="Carroll">Susan M. Carroll</name><name sortKey="Derose, Margaret L" sort="Derose, Margaret L" uniqKey="Derose M" first="Margaret L." last="Derose">Margaret L. Derose</name><name sortKey="Wahl, Geoffrey M" sort="Wahl, Geoffrey M" uniqKey="Wahl G" first="Geoffrey M." last="Wahl">Geoffrey M. Wahl</name></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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