Transposomics™ Research |
In Vivo Transposomics™ Research
Order
| Protocol
| Citations
| Related Products
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 in vivo transposition following
electroporation into living cells.**
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.
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
- Goryshin, I.Y. and Reznikoff , W.S. (1998) J. Biol. Chem. 273,
7367.
- Davies, D.R. et al. (2000) Science 289, 77.
- Shevchenko, Y. et al. (2002) Nucleic Acids Res. 30,
2469.
*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.
EZ-Tn5™ Transposon Construction Vectors
TransforMax™ EC100™-T1R Electrocompetent E. coli
TransforMax™ EC100™ Electrocompetent E. coli
TransforMax™ EC100™ Chemically Competent E. coli
[top of page]
|
