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.

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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.

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EZ-Tn5™ Transposase

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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.
4. Allsopp, L. P., et al. (2010) UpaH Is a Newly Identified Autotransporter Protein That Contributes to Biofilm Formation and Bladder Colonization by Uropathogenic Escherichia coli CFT073, Infect. Immun. 78 , 1659-1669.
5. Bian, Q. & Belmont, A. S. (2010) BAC TG-EMBED: one-step method for high-level, copy-number-dependent, position-independent transgene expression, Nucleic Acids Res. , gkq178.
6. Oh, J., et al. (2010) A universal TagModule collection for parallel genetic analysis of microorganisms, Nucleic Acids Res. , gkq419.
7. Yamanaka, K., et al. (2010) Mechanism of {varepsilon}-Poly-L-Lysine Production and Accumulation Revealed by Identification and Analysis of an {varepsilon}-Poly-L-Lysine-Degrading Enzyme, Appl. Envir. Microbiol. 76 , 5669-5675.
8. Yu, H. & Kim, K. S. (2010) Ferredoxin Is Involved in Secretion of Cytotoxic Necrotizing Factor 1 across the Cytoplasmic Membrane in Escherichia coli K1, Infect. Immun. 78 , 838-844.
9. Wei, Q., et al. (2009) dfrA27, a new integron-associated trimethoprim resistance gene from Escherichia coli, J. Antimicrob. Chemother. 63 , 405-406.
10. Zhou, L., et al. (2009) Transcriptional Regulation of the Escherichia coli Gene rraB, Encoding a Protein Inhibitor of RNase E, J. Bacteriol. 191 , 6665-6674.
11. Cho, K. H. & Caparon, M. G. (2008) tRNA Modification by GidA/MnmE Is Necessary for Streptococcus pyogenes Virulence: a New Strategy To Make Live Attenuated Strains, Infect. Immun. 76 , 3176-3186.
12. Murray, T. S. & Kazmierczak, B. I. (2008) Pseudomonas aeruginosa Exhibits Sliding Motility in the Absence of Type IV Pili and Flagella, J. Bacteriol. 190 , 2700-2708.
13. Reid, A. N., et al. (2008) Identification of Campylobacter jejuni Genes Contributing to Acid Adaptation by Transcriptional Profiling and Genome-Wide Mutagenesis, Appl. Envir. Microbiol. 74 , 1598-1612.
14. Kiyohara, Y. B., et al. (2006) The BMAL1 C terminus regulates the circadian transcription feedback loop, PNAS 103 , 10074-10079.

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For Research Use Only. Covered by issued and pending patents

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• 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.**

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.

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**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.

If you cannot find the answer to your problem then please contact us or telephone +44 (0)1954 210 200

If you cannot find the answer to your problem then please contact us or telephone +44 (0)1954 210 200