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
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.
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