RESEARCH ARTICLE


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



Shungo Kobori1, Yumi Ko2, Mikio Kato1, *
1 Department of Biological Science, Graduate School of Science, Osaka Prefecture University, 1-1 Gakuencho, Sakai 599-8531, Japan
2 Department of Earth and Life Science, College of Integrated Arts and Sciences, Osaka Prefecture University, 1-1 Gakuencho, Sakai 599-8531, Japan


Article Metrics

CrossRef Citations:
0
Total Statistics:

Full-Text HTML Views: 1716
Abstract HTML Views: 954
PDF Downloads: 266
Total Views/Downloads: 2936
Unique Statistics:

Full-Text HTML Views: 621
Abstract HTML Views: 629
PDF Downloads: 162
Total Views/Downloads: 1412



© Kobori et al.; Licensee Bentham Open.

open-access license: 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.

* Address correspondence to this author at the Department of Biological Science, Graduate School of Science, Japan; Tel/Fax: +81-72-254-9746; E-mail: mkato@b.s.osakafu-u.ac.jp,mikio_kato@mac.com


Abstract

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.

Keywords: Bent DNA, cloning vector, DNA curvature, IS10, mobile element.