《M1GS-HCV／C141核酶的构建及其体外抗病毒活性测定.》.pdf

李喜芳~M／IGS—HCVC／，核酶的构建及其体外抗病毒活性测定●髓 InterferODtreatmentincombinationwithribaviriniSusedasthefirst1ineclinicaltreatmentforHCV infection ． However． goodresponsetothistreatmenthasonlybeen observed in feW patientsandrepeatedrecurrencehasalsobeenreported frequently．Therefore，new antiviralagentsand therapiesare inurgentdemand．Here，wereportanewly constructed Escherichia coliRNaseP basedM 1GSribozymethatcanspecifically andeffi cientlytargetthecoregeneofHCV． The guidesequence(GS)ofthisMIGSwasdesignedaccordingtothesequenceofthecorecodingregionofHCVgenome．The GSwasthencovalentlylinkedtothe3’terminusofM 1RNA，thecatalyticsubunitofRNasePfrc．m EscherichiacoliThe ． specification ofthis sequence—specific ribozyme，M 1GS，was then examined using an in vitro cleavage assay ． The cytotoxicityanditsactivityininhibitionofHCV geneexpressionandviralproliferationwerefurtherstudiedinvivo ． Our resultsshow thatthereconstructedM IGSribozymedisplayedobviouscatalyticactivityincleavingtargetmRNAsrfagment invitro．Notablereduction intheexpressionofHCV coreproteinanda 1000．foldreduction inviralgrowth werealso observedin culturedHCV irifectedHuh7．5．1cellsexpressingthefunctionalM 1GSribozyme．Thisstudydemonstrateda directevidencefortheantiviralactivityofthecustomizedM 1GS—HCV／C】4】ribozyme andthusprovidedapromisingnew ， strategyofrclinicaltreatmentofHCV infection． Keywords： HCV,coregene，M1GSribozyme，antiviral 丙型肝炎是 由丙型肝炎病毒 (HepatitisC