1、Tab1.eSI.PrimersusedforthisstudyPrimerSequence5*to3DescriptionPCR(kb)aPUrPoSe阳JIiPFCatcccttgcctacctgaHankingregiondownstreamOfChVE1.1.RPb0.78CItVE1.I,1.阳JUPRTCCTGTGTGAAATTGTrATCCGCCCAACAAACCCAATCCACRankingregiond。WnWemOfChVEUFPCVfcXZdownrGICGrGAC1.GGGAAAACCcTGgcgtctgcaacgagtcagcACtankingregionupstre
2、amofChVEURP0.72VEZJdownRTcttcggcctattcatgcRankingregionupstreamofChVE1.IFPN1.C37GGAIGCCrcrGCTCGAAG17HygBsp1.itmarkerRP1.2MI3RAGCGGATAACAATrrCACACAGGAFPtoamp1.ifyHygBMI3FCGCCAGGGTnTCCCAGTCACGACRPtoamp1.ifyHfB1.8N1.C38CgttgcaagacctgcctgaaHygBsp1.itmarkerFPVttZupVERrccccrrGGGCRPdownstreamofChVE1.1.veri
3、ficationprimer,pairwithN1.C372.0ChVE1.IconfirmationVE7dwnVER2TGCTTGTGTCTCTGCCCCFPupstreamofChVE1.I.verificationPri1.Mr,;IirwithN1.C3825DWIOIAgcacgctcaacccaatgFPUpsirvainofChVEIJ.verificationPrimCr15ChVEIJcomnt-at沁nConfirinaiionChVEUR1.GcggtgggtaacaaagtgatC1.E1.codingregion.RPDW26GIcTreGGCrCCnrTCATCC
4、hVE1.ICodingregion.I：P24DWIo2GCATGGGAGCAGGAGATrACRPdownstreamofChVE1.1.verificationprimerCh1.AE1.WXAaagatgcagatgggtggacHankingregionupstreamofCMt71.P0.77Ch1.A1.ICh1.AE1.RP1.CGACCXCTrCCCGTCCCctgtgtgaaattgttatccGCTthinkingregionupstreamof(h1.1.IRPCh1.AE1.FP1.CccgttccatgccagcaaagTcgtgactgggaaaaccctgGCG
5、I1.ankingregiondownstreamo(Ch1.AE1.FP0.63CMt7RP2CeCACGCTAGAAACeTrCAGf1.ankingregiondownstreamCh1.AEIRPC1.EiGgatgggtcccagaggtgupstreamofChtAEIverificationrinr.FP.pairwithN1.C3710CMAEIconfirmationCh1.AEIDAtactgcctgatccgtgtccdownstreamofCh1.AEIverificationprimer,P.pairwithN1.C382.7DW27OAAGATCACTGGAzXCA
6、ACTGGCATGCCCACGCAGAAACCTTCAGRptoump1.ifyCh1.AEIsequencewithPirpCtai1.forCompkmenUUionpairwithChIAEim2.7CMAEIcomp1.ementationDW69CTGCCAG11GCCAGIG1.PrpCpromoter2.7DW70AcctctaaacaagtgtacctGRPTrpCterminatorDW27IGCACAGGTACACTTGTrTAGAggtggcaaagtggctcggcatFPcoumpIifyCAf7ck)wns(rvmankingsequencewithTirpCtai
7、1.forcomp1.ementation0.7DW272GccaaaacaacttcaccacRPtoamp1.ifyCMEdownsirca11f1.unkingsequeefcompkmcntaiionTtrpCcndGtgaatgctccgtaacacccAATACFPUCATPH.TcnniiiatorOfTrpC1.2Ch1.AEIcomp1.ementationCOnfimuIiOnDW273TGre1.1.GTGICrGnCeTrCGRPtoraxftrni(ionCh1.AEntaionCh1.EIFGgccgctggtatcatggcttCCG11bpdownsreamof
8、startcodonofCh1.AEI1.2Ch1.AE1RAgcatcggcccagtaacatcAAACnearstopCOdOnofCh1.AEIDW28TGCAggatccATGTCCAACATT11GIGVcAOEFPatATGwithAomH1.cutsite2.8ChVE1.IOEcp1.asmidSInIC1.ionDW29TCGGaagcttGTFTCACCTTTGGGGTAATCVcAOERPat+287bpwithHinMWcutsiteDW30CGAGggatccATGGAAAATGGCCGCTGG1.aCAOEFPatATGwith80mHIcusite2.0Ch1.
9、AE1.OEp1.asmidconstmctioDW3ICGTcaagcuCTATACCGTCATGCCGAG14eAOERP加376bwithHiTtdII1.cutsiteDW38Agatggtcaacgctgcttacpc1.APrOmeHcrPnmCrforconfirmationOfoEstrains.41.bpawayfromv1.1.site2.9ChVE1.JOEconfirmationDW39CGAGI11GGGATi1.GAG94bpawayfromDW29forConfirnuiiKmofChVEIJOEstrainswithDV38DW40Actctagaacatcct
10、cgacg21.5bpawayfromDW31forconfirmationofCMAE1.OEstrainswithDW382.2Ch1.AEIOEconfirmationDW4IGcTGGTTGGAGAAGTATGAGTreha1.oseRPforconfirmationofPromoter1integration1.0ChVEIJandCh1.AEIOEconfirmationDW42AeGACCCIGAATGACAAGPcIAPrOnW1.erRPforConfirnKiiMinofPromoterIintegration,pairwithDV41PCRIkb)=PCRproductIcngihinkb:hFP=forwardprimer;cRP-revcneprimerd：=genede1.etion;OE=Ovcrcxprcssion
