title: A set of mini-Mu transposons for versatile cloning of circular DNA and novel dual-transposon strategy for increased efficiency.
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.
authors.affiliation: Division of Genetics and Physiology, Department of Biology, University of Turku, Vesilinnantie 5, FI-20500 Turku, Finland.
authors.forename: Elsi
authors.initials: E
authors.lastname: Pulkkinen
authors.affiliation: Division of Genetics and Physiology, Department of Biology, University of Turku, Vesilinnantie 5, FI-20500 Turku, Finland.
authors.forename: Saija
authors.initials: S
authors.lastname: Haapa-Paananen
authors.affiliation: Institute of Biotechnology, Viikki Biocenter, P.O. Box 56, Viikinkaari 9, FI-00014, University of Helsinki, Helsinki, Finland.
authors.forename: Hilkka
authors.initials: H
authors.lastname: Turakainen
authors.affiliation: Division of Genetics and Physiology, Department of Biology, University of Turku, Vesilinnantie 5, FI-20500 Turku, Finland; Institute of Biotechnology, Viikki Biocenter, P.O. Box 56, Viikinkaari 9, FI-00014, University of Helsinki, Helsinki, Finland. Electronic address: harri.savilahti@utu.fi.
authors.forename: Harri
authors.initials: H
authors.lastname: Savilahti
mesh_terms: D003001:Cloning, Molecular / Q000379:methods*; D004251:DNA Transposable Elements / Q000235:genetics*; D004270:DNA, Circular / Q000235:genetics*; D004926:Escherichia coli / Q000235:genetics; D018741:Replication Origin / Q000235:genetics; D019895:Transposases / Q000378:metabolism*
chemical_list: D004251:DNA Transposable Elements; D004270:DNA, Circular; C114697:mu transposase; D019895:Transposases
keywords: Autointegration; Cloning of circular DNA; Origin of replication; Phage Mu; Replication-deficient