Applications

• InvitroinsertionofanEZ-Tn5TransposonintoDNAclonedinvectors,suchasplasmids,fosmids,cosmids,orBACsaswellasinvitroinsertionintolinearDNA.*
• PreparationofEZ-Tn5Transposomesforinvivotranspositionfollowingelectroporationintolivingcells.**

EZ-Tn5™TransposaseisahyperactiveformofTn5transposase.¹Thehighlypurified,single-subunitenzymecanbeusedtorandomlyinsert(transposeor"hop")anyEZ-Tn5TransposonintoanytargetDNAinvitrowithanefficiencyupto>10⁶insertionclonesperstandardreaction.WhenincubatedwithanEZ-Tn5TransposonintheabsenceofMg²⁺,astableEZ-Tn5Transposome™complexisformed.TheTransposomeissostablethatitcanbeelectroporatedintolivingcells.Onceinthecell,theTransposomeisactivatedbyintracellularMg²⁺andtheEZ-Tn5Transposoncomponentisrandomlyinsertedintothehost'sgenomicDNA.Thethree-dimensionalstructureofaTransposomecomplexhasbeenelucidated.²

AtypicalEZ-Tn5transpositionreactionrequiresfourcomponents:(1)theEZ-Tn5Transposase;(2)anEZ-Tn5Transposon;(3)atargetDNA;and(4)thepresenceofMg²⁺.ThehighlyrandominsertionofanEZ-Tn5TransposonintothetargetDNA³proceedsbyacut-and-pastemechanism,¹catalyzedbytheEZ-Tn5Transposase,andresultsina9-bpduplicationoftargetDNAsequenceimmediatelyadjacenttobothendsoftheTransposon.

Benefits

• SpecificallyanduniquelyrecognizestheOuterEndsequencesofnaturallyoccurringTn5andmini-Tn5transposonsandthehyperactiveMosaicEndsofEZ-Tn5Transposons.

References

1. Goryshin,I.Y.andReznikoff,W.S.(1998)J.Biol.Chem.273,7367.
2. Davies,D.R.etal.(2000)Science289,77.
3. Shevchenko,Y.etal.(2002)NucleicAcidsRes.30,2469.

*Coveredbyissuedand/orpendingpatents,exclusivelylicensedorassignedtoEpicentre®(anIllumina®Company).
**TheuseofTransposome™complexesforinvivoinsertionofatransposon,including,butnotlimited,toHyperMu™andEZ-Tn5™Transposome™complexes,iscoveredbyU.S.PatentNo.6,159,736andrelatedpatentsandpatentapplications,exclusivelylicensedtoEpicentre®(anIllumina®Company).