Pre-madelentiviralparticlesbicistronicallyexpresstwoindividualproteins:fireflyluciferaseandnuclearpermeableCRE recombinase,underthesamesuCMVpromoter.ARFP-PuromycinfusiondualarkerwasexpressedunderRSVpromoter,whichallowtoselectthepositivetransducedstablecellsforlongtermcultureviaRFPcellsortingorviaantibioticselection.
BothlucifeaseandCREarehighlyactive,bestforbothenzymes'assaysInvivoorInvitro.CREprotein,frombacteriophageP1,catalyzesrecombinationbetween34basepairtargetsequences, namedloxsites.CRElentiviralparticles can catalyze thejointingeventInVivo inthepresenceof loxsites.This nuclearpermeant CRE(NLS-CRE) increasestherecombinationeventsinvivo. PurifiedCREenzymecanjointindividualplasmidscontainingloxsites.
LuciferaeandNLS-CRE areconstitutivelyexpressedinhighlevel,andoptionally,alsocanbeusedastetracyclineinducIBLeexpression asdesired. LuciferaseandRFPprovide theconvenient,realtimemonitoring signals. Seeproductmanualfordetails(.pdf). Amount: 200ul/pervial,at 1x107IFU/mlinDMEMmediumwith10%FBS. Cat#:LVP414
