Selective excision of the centromere chromatin complex from Saccharomyces cerevisiae

Selective excision of the centromere chromatin complex from Saccharomyces cerevisiae

We have taken advantage of the known structural parameters associated with centromere DNA in vivo to construct a CEN fragment that can be selectively excised from the chromatin DNA with restriction endonucleases. CEN3 DNA is organized in chromatin such that a 220-250- bp region encompassing the elem...

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Format: Online
Language:English
Published: The Rockefeller University Press 1988
Online Access:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2115176/
id pubmed-2115176
recordtype oai_dc
spelling pubmed-21151762008-05-01 Selective excision of the centromere chromatin complex from Saccharomyces cerevisiae Articles We have taken advantage of the known structural parameters associated with centromere DNA in vivo to construct a CEN fragment that can be selectively excised from the chromatin DNA with restriction endonucleases. CEN3 DNA is organized in chromatin such that a 220-250- bp region encompassing the elements of centromere homology is resistant to nuclease digestion. Restriction enzyme linkers encoding the Bam HI- recognition site were ligated to a 289 base pair DNA segment that spans the 220-250-bp protected core (Bloom et al., 1984). Replacement of this CEN3-Bam HI linker cassette into a chromosome or plasmid results in formation of a complete structural and functional centromeric unit. A centromere core complex that retains its protected chromatin conformation can be selectively excised from intact nuclei by restriction with the enzyme Bam HI. The centromeric protein-DNA complex is therefore not dependent upon the intact torsional constrains on linear chromosomes for its structural integrity. Isolation of this complex provides a novel approach to characterizing authentic centromeric proteins bound to DNA in their native state. The Rockefeller University Press 1988-07-01 /pmc/articles/PMC2115176/ /pubmed/2839524 Text en This article is distributed under the terms of an Attribution–Noncommercial–Share Alike–No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms). After six months it is available under a Creative Commons License (Attribution–Noncommercial–Share Alike 4.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/4.0/).
repository_type Open Access Journal
institution_category Foreign Institution
institution US National Center for Biotechnology Information
building NCBI PubMed
collection Online Access
language English
format Online
title Selective excision of the centromere chromatin complex from Saccharomyces cerevisiae
spellingShingle Selective excision of the centromere chromatin complex from Saccharomyces cerevisiae
title_short Selective excision of the centromere chromatin complex from Saccharomyces cerevisiae
title_full Selective excision of the centromere chromatin complex from Saccharomyces cerevisiae
title_fullStr Selective excision of the centromere chromatin complex from Saccharomyces cerevisiae
title_full_unstemmed Selective excision of the centromere chromatin complex from Saccharomyces cerevisiae
title_sort selective excision of the centromere chromatin complex from saccharomyces cerevisiae
description We have taken advantage of the known structural parameters associated with centromere DNA in vivo to construct a CEN fragment that can be selectively excised from the chromatin DNA with restriction endonucleases. CEN3 DNA is organized in chromatin such that a 220-250- bp region encompassing the elements of centromere homology is resistant to nuclease digestion. Restriction enzyme linkers encoding the Bam HI- recognition site were ligated to a 289 base pair DNA segment that spans the 220-250-bp protected core (Bloom et al., 1984). Replacement of this CEN3-Bam HI linker cassette into a chromosome or plasmid results in formation of a complete structural and functional centromeric unit. A centromere core complex that retains its protected chromatin conformation can be selectively excised from intact nuclei by restriction with the enzyme Bam HI. The centromeric protein-DNA complex is therefore not dependent upon the intact torsional constrains on linear chromosomes for its structural integrity. Isolation of this complex provides a novel approach to characterizing authentic centromeric proteins bound to DNA in their native state.
publisher The Rockefeller University Press
publishDate 1988
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2115176/
_version_ 1611413665050787840

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