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Nucleic Acids Research, 2000, Vol. 28, No. 5 1067-1077
© 2000 Oxford University Press

A simple in vitro Tn7-based transposition system with low target site selectivity for genome and gene analysis

Matthew C. Biery, Fiona J. Stewart1, Anne E. Stellwagen, Elisabeth A. Raleigh1 and Nancy L. Craig*

Howard Hughes Medical Institute, Department of Molecular Biology and Genetics, Johns Hopkins University School of Medicine, 725 North Wolfe Street, Baltimore, MD 21205, USA and 1New England Biolabs Inc., 32 Tozer Road, Beverly, MA 01915, USA

A robust Tn7-based in vitro transposition system is described that displays little target site selectivity, allowing the efficient recovery of many different transposon insertions in target DNAs ranging from small plasmids to cosmids to whole genomes. Two miniTn7 derivatives are described that are useful for the analysis of genes: one a derivative for making translational and transcriptional target gene fusions and the other a derivative that can generate 15 bp (5 amino acid) insertions in target DNAs (proteins).

* To whom correspondence should be addressed. Tel: +1 410 955 3933; Fax: +1 410 955 0831; Email: ncraig@jhmi.edu Present address: Anne E. Stellwagen, Fred Hutchinson Cancer Research Center, 1100 Fairview Avenue North, Mailstop A3-025, Seattle, WA 98109, USA


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