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农杆菌Thegenetictransformationisamultiple-stepprocesswhichincludes:(1)Agrobacterium-hostrecognitionandattachment;(2)sensingofspecificplantsignals;(3)signaltransductionandvirulenceactivation;(4)generationofamobilecopyofT-DNA,theT-strand;(5)generationoftheVirB-VirD4transportercomplex,andtransportofT-strandsandVirproteinsintothehostcellcytoplasm;(6)formationofthematureT-complex;(7)T-complexmovementthroughthecytoplasmandnuclearimport;(8)T-complexdisassociationandT-DNAintegrationintothehostcellgenome.(IM,bacterialinnermembrane;NPC,nuclearporecomplex;OM,bacterialoutermembrane;PP,bacterialperiplasm).Ti质粒SouthernblotRealtimePCRDNAfiberFISH质粒拯救plasmidrescue反式PCRinversePCRSitefindingPCR基因整合方式同源重组锌指蛋白核酸酶ZFN重组酶介导的盒式交换RMCE