Description:
IMPACTCitations
pTYB21isanE.colicloningandexpressionvector(7514bp)usedintheIMPACT?ProteinPurificationSystemwhichallowstheoverexpressionofatargetproteinasafusiontoaself-cleavableaffinitytag(1,2).ItisanN-terminalfusionvectordesignedforin-frameinsertionofatargetgeneintothepolylinker,downstreamoftheinteintag(theSceVMAintein/chitinbindingdomain,55kDa)(3,4).ThisallowstheN-terminusofthetargetproteintobefusedtotheinteintag.Theself-cleavageactivityoftheinteinallowsthereleaseofthetargetproteinfromthechitin-boundinteintag,resultinginasinglecolumnpurificationofthetargetprotein.ThisvectorcanbeusedinconjuctionwithaC-terminalfusionvectortotestwhichfusionconstruction(N-terminalorC-terminal)maximizestheexpressionandyieldofatargetprotein.ForthefusionoftheC-terminusofthetargetproteintotheinteintag,usepTXB1(NEB?#N6707),pTXB3(NEB#N6708),pTYB1(NEB#N6701),pTYB2(NEB#N6702),pTYB3(NEB#N6703)orpTYB4(NEB#N6704).