A Target Site for Spontaneous Insertion of IS10 Element in pUC19 DNA Located within Intrinsically Bent DNA

Residual insertion sequence elements (IS elements) in Escherichia coli strains that are commonly used for DNA cloning are known to cause cloning artifacts by transposing themselves into the recombinant DNA fragments. In such cases, chance insertion of IS elements may occur at integration sites in th...

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Detalles Bibliográficos
Autores principales: Kobori, Shungo, Ko, Yumi, Kato, Mikio
Formato: Texto
Lenguaje:English
Publicado: Bentham Open 2009
Materias:
Article
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2758499/
https://www.ncbi.nlm.nih.gov/pubmed/19812719
http://dx.doi.org/10.2174/1874285800903010146
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author Kobori, Shungo
Ko, Yumi
Kato, Mikio
author_facet Kobori, Shungo
Ko, Yumi
Kato, Mikio
author_sort Kobori, Shungo
collection PubMed
description Residual insertion sequence elements (IS elements) in Escherichia coli strains that are commonly used for DNA cloning are known to cause cloning artifacts by transposing themselves into the recombinant DNA fragments. In such cases, chance insertion of IS elements may occur at integration sites in the cloning targets, which in the case of the IS10 element is a 9-bp consensus sequence. We report here that the integration of IS10-related DNA sequences into the pUC19 cloning vector and its derivative occurred with considerable frequency in E. coli strains JM107 and DH10B, with duplication of a 9-bp segment (TCTAAAGTA). Notably, native polyacrylamide gel electrophoresis revealed that intrinsically bent DNA flanks the insertion site.
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spelling pubmed-27584992009-10-07 A Target Site for Spontaneous Insertion of IS10 Element in pUC19 DNA Located within Intrinsically Bent DNA Kobori, Shungo Ko, Yumi Kato, Mikio Open Microbiol J Article Residual insertion sequence elements (IS elements) in Escherichia coli strains that are commonly used for DNA cloning are known to cause cloning artifacts by transposing themselves into the recombinant DNA fragments. In such cases, chance insertion of IS elements may occur at integration sites in the cloning targets, which in the case of the IS10 element is a 9-bp consensus sequence. We report here that the integration of IS10-related DNA sequences into the pUC19 cloning vector and its derivative occurred with considerable frequency in E. coli strains JM107 and DH10B, with duplication of a 9-bp segment (TCTAAAGTA). Notably, native polyacrylamide gel electrophoresis revealed that intrinsically bent DNA flanks the insertion site. Bentham Open 2009-09-25 /pmc/articles/PMC2758499/ /pubmed/19812719 http://dx.doi.org/10.2174/1874285800903010146 Text en © Kobori et al.; Licensee Bentham Open. http://creativecommons.org/licenses/by-nc/3.0/ This is an open access article licensed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0/) which permits unrestricted, non-commercial use, distribution and reproduction in any medium, provided the work is properly cited.
spellingShingle Article
Kobori, Shungo
Ko, Yumi
Kato, Mikio
A Target Site for Spontaneous Insertion of IS10 Element in pUC19 DNA Located within Intrinsically Bent DNA
title A Target Site for Spontaneous Insertion of IS10 Element in pUC19 DNA Located within Intrinsically Bent DNA
title_full A Target Site for Spontaneous Insertion of IS10 Element in pUC19 DNA Located within Intrinsically Bent DNA
title_fullStr A Target Site for Spontaneous Insertion of IS10 Element in pUC19 DNA Located within Intrinsically Bent DNA
title_full_unstemmed A Target Site for Spontaneous Insertion of IS10 Element in pUC19 DNA Located within Intrinsically Bent DNA
title_short A Target Site for Spontaneous Insertion of IS10 Element in pUC19 DNA Located within Intrinsically Bent DNA
title_sort target site for spontaneous insertion of is10 element in puc19 dna located within intrinsically bent dna
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2758499/
https://www.ncbi.nlm.nih.gov/pubmed/19812719
http://dx.doi.org/10.2174/1874285800903010146
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