A1 Refereed original research article in a scientific journal
A set of mini-Mu transposons for versatile cloning of circular DNA and novel dual-transposon strategy for increased efficiency
Authors: Pulkkinen E, Haapa-Paananen S, Turakainen H, Savilahti H
Publisher: ACADEMIC PRESS INC ELSEVIER SCIENCE
Publication year: 2016
Journal: Plasmid
Journal name in source: PLASMID
Journal acronym: PLASMID
Volume: 86
First page : 46
Last page: 53
Number of pages: 8
ISSN: 0147-619X
eISSN: 1095-9890
DOI: https://doi.org/10.1016/j.plasmid.2016.07.002(external)
Abstract
Mu transposition-based cloning of DNA circles employs in vitro transposition reaction to deliver both the plasmid origin of replication and a selectable marker into the target DNA of interest. We report here the construction of a platform for the purpose that contains ten mini-Mu transposons with five different replication origins, enabling a variety of research approaches for the discovery and study of circular DNA. We also demonstrate that the simultaneous use of two transposons, one with the origin of replication and the other with selectable marker, is beneficial as it improves the cloning efficiency by reducing the fraction of autointegration-derived plasmid clones. The constructed transposons now provide a set of new tools for the studies on DNA circles and widen the applicability of Mu transposition based approaches to clone circular DNA from various sources. (C) 2016 Elsevier Inc. All rights reserved.
Mu transposition-based cloning of DNA circles employs in vitro transposition reaction to deliver both the plasmid origin of replication and a selectable marker into the target DNA of interest. We report here the construction of a platform for the purpose that contains ten mini-Mu transposons with five different replication origins, enabling a variety of research approaches for the discovery and study of circular DNA. We also demonstrate that the simultaneous use of two transposons, one with the origin of replication and the other with selectable marker, is beneficial as it improves the cloning efficiency by reducing the fraction of autointegration-derived plasmid clones. The constructed transposons now provide a set of new tools for the studies on DNA circles and widen the applicability of Mu transposition based approaches to clone circular DNA from various sources. (C) 2016 Elsevier Inc. All rights reserved.
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