EZ-Tn5™ Transposase is a hyperactive form of Tn5 transposase.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™ 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 DNA3 proceeds by a cut-and-paste mechanism,1 catalyzed
by the EZ-Tn5 Transposase, and results in a 9-bp duplication of target DNA
sequence immediately adjacent to both ends of the Transposon.
*Covered by issued and/or pending patents.
**The use of Transposome™ complexes for in vivo insertion 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.