 Wysłany: Pon 11:09, 04 Kwi 2011 Temat postu: Tn5 transposon inserted into AcMNPV p95 gene does |
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| Tn5 transposon inserted p95 gene does not affect the replication of AcMNPV
weversL, IatrouK. Thep95GeneofBombyxmorinuclearpolyhedrosisvirus: temporalexpressionandfunctionalproperties [J]. JVirol ,1998,72:4789-4797. [12] RussellRLQ, RohrmarmGF. CharacterizationofP91, aproteinassociatedwithvirionsofan0rapseudot-sugatabaculovirus [J]. Virology ,1997,belstaff outlet,233:210-223. InsertionalDisruptionofAcMNPVp95GenedidnotAffectVirusReplicationinSf9CellsYINJuan, HUZhi-peng, SONGDa-xin, ZHONGJiang (DepartmentofMicrobiologyandMicrobialEngineering, SchoolofLSciences, FudanUniversity, Shanghai200433, China) Abstract: Areplication-competentmutantvirus, AcApra41, wasisolatedfromalibraryofAcM = NPVrandominsertionaImutantsconstructedwithTn5transposonmediatedmutagenesis. TheTn5transposonwasdetenninedtobelocatedinthep95geneofAcMNPVgenome. ToexcludethepossibilitythatAcApra41containedothermutations, arecombinantvirus, AcGFP-P95in, whichhadtargetedinsertionofTn5inp95geneinexactlythesamewayasinAcApra41, wasconstructedbyhomologousrecombination. TheinsertionofTn5transposonwasconfirmedwithPcR. Theinsertionoftransposonre-suitedintheappearanceofatruncatedformofP95proteinof55kuincellsinfectedwithAc (FP-P95inasshownbyWesternblotusingantibodyagainsttheN-terminalpartofP95protein, incontrastwiththefulllengthP95proteinof95kuincellsinfectedwithwildtypevirus.VirusreplicationCHIVeandfluorescentmicroscopicobservationindicatedthatAcGFP-P95inreplicatedandexpressedviralgenesnormallyaswildtypevirusinSf9cells.Theresultsindicatedthatafull-lengthP95proteinwasnotessentiaIforvirusreplication.Keywords: Baculovirus; mutants; replication; geneexpression |
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