EZ-Tn5™ Transposase

For Research Use Only. Covered by issued and pending patents

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Last modified: 2008-07-15 14:57:15

EZ-Tn5™ Transposase is a hyperactive form of Tn5 transposase described by Goryshin and Reznikoff.¹ The highly purified, single-subunit enzyme can be used to randomly insert (transpose or 'hop') any EZ-Tn5™ Transposon into any target DNA in vitro with an efficiency up to >106 insertion clones per standard reaction. When incubated with an EZ-Tn5™ Transposon in the absence of Mg²+, a stable EZ-Tn5™ Transposome™ is formed.
The Transposome™ complex 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. EZ-Tn5™ Transposase
2. EZ-Tn5™ Transposon
3. a target DNA
4. Mg²+.

Insertion of an EZ-Tn5™ Transposon into the target DNA proceeds by a cut and paste mechanism,¹ catalysed by the EZ-Tn5™ Transposase, and results in a 9-bp duplication of target DNA sequence immediately adjacent to both ends of the Transposon.

Last modified: 2008-05-14 16:46:46

Applications

• in vitro insertion of an EZ-Tn5™ Transposon into DNA cloned in vectors, such as plasmids, fosmids, cosmids or BACs as well as in vitro insertion into linear DNA
• Preparation of EZ-Tn5™ Transposomes for delivery into living cells by electroporation

Benefits

• Hyperactive form of Tn5 transposase
• Specifically and uniquely recognises the Outer End sequences of naturally occurring Tn5 and mini-Tn5 transposons and the hyperactive Mosaic Ends

Last modified: 2008-05-14 16:46:46

Protocols for: EZ-Tn5™ Transposase

(catalogue number TNP92110)

Please note: all protocols off site are the responsibility of the products supplier

Last modified: 2008-05-14 16:46:46

References

1. Goryshin, I.Y. and Reznikoff, W.S. (1998) J. Biol. Chem. 273, 7367.
2. Davies, D.R. et al. (2000) Science 289 (5476), 77.
3. Shevchenko, Y. et al. (2002) Nucl. Acids Res. 30, 2469.

Last modified: 2008-05-14 16:46:46