A Target Site for Spontaneous Insertion of IS10 Element in pUC19 DNA Located within Intrinsically Bent DNA
Shungo Kobori1, Yumi Ko2, Mikio Kato1, *
Identifiers and Pagination:Year: 2009
First Page: 146
Last Page: 150
Publisher ID: TOMICROJ-3-146
Article History:Received Date: 18/8/2009
Revision Received Date: 25/8/2009
Acceptance Date: 27/8/2009
Electronic publication date: 25/9/2009
Collection year: 2009
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.
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.