EZ-Tn5- Transposase

• In vitroinsertion of an EZ-Tn5 Transposon into DNA cloned in vectors, such as plasmids, fosmids, cosmids, or BACs as well asin vitroinsertion into linear DNA.*
• Preparation of EZ-Tn5 Transposomes forin vivotransposition following electroporation into living cells.**

EZ-Tn5™ Transposase is a hyperactive form of Tn5 transposase.1The
highly purified, single-subunit enzyme can be used to randomly insert (transpose
or “hop”) any EZ-Tn5 Transposon into any target DNAin vitrowith
an efficiency up to >106insertion clones per standard reaction.
When incubated with an EZ-Tn5 Transposon in the absence of Mg2+,
a stable EZ-Tn5 Transposome™ complex is formed. The Transposome is so
stable that it can be electroporated into living cells. Once in the cell, the
Transposome is activated by intracellular Mg2+and the EZ-Tn5 Transposon
component is randomly inserted into the host’s genomic DNA. The three-dimensional
structure of a Transposome complex has been elucidated.2

A typical EZ-Tn5 transposition reaction requires four components: (1) the
EZ-Tn5 Transposase; (2) an EZ-Tn5 Transposon; (3) a target DNA; and (4) the
presence of Mg2+. The highly random insertion of an EZ-Tn5 Transposon
into the target DNA3proceeds by a cut-and-paste mechanism,1catalyzed
by the EZ-Tn5 Transposase, and results in a 9-bp duplication of target DNA
sequence immediately adjacent to both ends of the Transposon.

Benefits

• Specifically and uniquely recognizes the Outer End sequences of naturally occurring Tn5 and mini-Tn5 transposons and the hyperactive Mosaic Ends of EZ-Tn5 Transposons.

References

1. Goryshin, I.Y. and Reznikoff , W.S. (1998)J. Biol. Chem.273, 7367.
2. Davies, D.R.et al.(2000)Science289, 77.
3. Shevchenko, Y.et al.(2002)Nucleic Acids Res.30, 2469.

*Covered by issued and/or pending patents.**The use of Transposome™ complexes forin vivoinsertion of a
transposon, including, but not limited, to HyperMu™ and EZ-Tn5™ Transposome™ complexes,
is covered by U.S. Patent No. 6,159,736 and related patents and patent applications,
exclusively licensed to Epicentre.

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