Rapid generation of long tandem DNA repeat arrays by homologous recombination in yeast to study their function in mammalian genomes

We describe here a method to rapidly convert any desirable DNA fragment, as small as 100 bp, into long tandem DNA arrays up to 140 kb in size that are inserted into a microbe vector. This method includes rolling-circle phi29 amplification (RCA) of the sequence in vitro and assembly of the RCA produc...

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Detalles Bibliográficos
Autores principales: Noskov, Vladimir N, Lee, Nicholas CO, Larionov, Vladimir, Kouprina, Natalay
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2011
Materias:
Methodology
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3200152/
https://www.ncbi.nlm.nih.gov/pubmed/21982381
http://dx.doi.org/10.1186/1480-9222-13-8
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author Noskov, Vladimir N
Lee, Nicholas CO
Larionov, Vladimir
Kouprina, Natalay
author_facet Noskov, Vladimir N
Lee, Nicholas CO
Larionov, Vladimir
Kouprina, Natalay
author_sort Noskov, Vladimir N
collection PubMed
description We describe here a method to rapidly convert any desirable DNA fragment, as small as 100 bp, into long tandem DNA arrays up to 140 kb in size that are inserted into a microbe vector. This method includes rolling-circle phi29 amplification (RCA) of the sequence in vitro and assembly of the RCA products in vivo by homologous recombination in the yeast Saccharomyces cerevisiae. The method was successfully used for a functional analysis of centromeric and pericentromeric repeats and construction of new vehicles for gene delivery to mammalian cells. The method may have general application in elucidating the role of tandem repeats in chromosome organization and dynamics. Each cycle of the protocol takes ~ two weeks to complete.
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spelling pubmed-32001522011-10-25 Rapid generation of long tandem DNA repeat arrays by homologous recombination in yeast to study their function in mammalian genomes Noskov, Vladimir N Lee, Nicholas CO Larionov, Vladimir Kouprina, Natalay Biol Proced Online Methodology We describe here a method to rapidly convert any desirable DNA fragment, as small as 100 bp, into long tandem DNA arrays up to 140 kb in size that are inserted into a microbe vector. This method includes rolling-circle phi29 amplification (RCA) of the sequence in vitro and assembly of the RCA products in vivo by homologous recombination in the yeast Saccharomyces cerevisiae. The method was successfully used for a functional analysis of centromeric and pericentromeric repeats and construction of new vehicles for gene delivery to mammalian cells. The method may have general application in elucidating the role of tandem repeats in chromosome organization and dynamics. Each cycle of the protocol takes ~ two weeks to complete. BioMed Central 2011-10-07 /pmc/articles/PMC3200152/ /pubmed/21982381 http://dx.doi.org/10.1186/1480-9222-13-8 Text en Copyright ©2011 Noskov et al; licensee BioMed Central Ltd. http://creativecommons.org/licenses/by/2.0 This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Methodology
Noskov, Vladimir N
Lee, Nicholas CO
Larionov, Vladimir
Kouprina, Natalay
Rapid generation of long tandem DNA repeat arrays by homologous recombination in yeast to study their function in mammalian genomes
title Rapid generation of long tandem DNA repeat arrays by homologous recombination in yeast to study their function in mammalian genomes
title_full Rapid generation of long tandem DNA repeat arrays by homologous recombination in yeast to study their function in mammalian genomes
title_fullStr Rapid generation of long tandem DNA repeat arrays by homologous recombination in yeast to study their function in mammalian genomes
title_full_unstemmed Rapid generation of long tandem DNA repeat arrays by homologous recombination in yeast to study their function in mammalian genomes
title_short Rapid generation of long tandem DNA repeat arrays by homologous recombination in yeast to study their function in mammalian genomes
title_sort rapid generation of long tandem dna repeat arrays by homologous recombination in yeast to study their function in mammalian genomes
topic Methodology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3200152/
https://www.ncbi.nlm.nih.gov/pubmed/21982381
http://dx.doi.org/10.1186/1480-9222-13-8
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