重组蛋白在植物体中的表达及其应用①

2004,21(1):101112ChineseBulletinofBotanyAuthorforcorrespondence.1963119962002302002-11-102003-05-11110034RecombinantProteinExpressioninPlantsandItsApplicationHAOLinXUXinWANGZun-Sheng(DepartmentofBiology,ShenyangNormalUniversity,Shenyang110034)AbstractStudiesontransgenicplantsasthebioreactortoproducepharmaceuticalandcommericialvaluableproteinshasbeenadvancedforoveradecade.Somerecombinantproteinsproducedinplantsarenowinclinicaltrialsandhaveapromisefuture.Anotherexpressedproteinsinplantsareagronomicallyvaluableandcanmediateresistancetopathogeninfections,socouldbeanalternativemethodinmolecularbreedingofplants.Inthisreview,weintroducesomeadvancesinthisfield.KeywordsRecombinantprotein,Transgenicplant,Plantibody,Pathogenresistance,Immunomodulation10(Hiattetal,1989;Higoetal,1993;Stögeretal,2002)(1)(LarrickandThomas,2001)234(5)(6)(FischerandEmans,2000)(7)(8)1plantibody1989(Hiattetal,1989),(Maetal,1995)(vanEngelenetal,1994)(Maetal,1995;1998)Fab(fragmentantigenbinding)(DeNeveetal,1993)scFv(singlechainantibodyfragment)(Owenetal,1992)scFv(Fischeretal,1999)0.5%~2%TSP()(ConradandFiedler,1998)1.1mRNAcDNAPCRFabscFv(Birdetal,1988),,scFv(Danielletal,2001),IgG/A(Maetal,1995)(Vaqueroetal,1999)phagedisplay(McCaffertyetal,1990)(BoonhamandBarker,1998;Harperetal,1999;Griepetal,2000)(DeJaegeretal,1998)1.2(Vaqueroetal,1999)(FischerandEmans,2000)3DNA(Christou,1993)(Kapilaetal,1996)(Scholthofetal,1996;Kumagaietal,2000)1.3CaMV35SFabNsecretorypathwayYuanetal,1997;ConradandFiedler,1998scFvNC4KDELscFvapoplastDaniell,1999(recombinantantibodies,rAbs)scFv(Schillbergetal,1999)scFvframworkregionKnappikandPlückthun,1995BtCRY1TSP47%DeCosa,2001KimandMayfield,1997;Staub,2000,(positioneffect),Danielletal,2001,Daniell,199922.11(Larricketal,1998;Maetal,1998;Danielletal,2001)rAbsrAbs(Danielletal,2001)(glycosylation)(Chargelegueetal,2000;Bakkeretal,2001)N-CKDEL-1,4-galactosyltra-nsferase(Bakkeretal,2001)2.2Langridge(2000)Ediblevaccines(ediblevaccines)2(Castanon,1999;Kapustaetal,1999;Tubolyetal,2000)(Danielletal,2001)polyA(Richteretal,2000)(Tubolyetal,2000)(Danielletal,2001)1Table1PlantibodiesandapplicationsApplicationAntibodytypeAntigenExpressionsystemReferencesSigA*Maetal,1998DentalcariesStreptococcalNicotianaantigenortabacumIgGIgGKhoudietal,1999DiagnosticAnti-humanIgGAlfalfascFvStögeretal,2000CancertreatmentCarcinoembryonicantigenWheat,RiceBscFvMcCormickB-celllymphomaIdiotypevaccineNicotianaetal,1999treatmentbenthamianaIgGVerchetal,1998ColoncancerSurfaceantigenNicotianatreatmentbenthamianaIgGWigdorovitzHerpestreatmentHerpessimplexvirus2Soybeanetal,1999*IgA(secretoryIgA)(Richteretal,2000;Tubolyetal,2000)2.33hirudinGiddings,20013TSP1%(Kusnadietal,1997,32Table2VaccinesexpressedbytransgenicplantsSourceofpurposegenesProteinorpeptideExpressionImmunogenicityReferencesandtargetspeciesforexpressedsystemcapacitythevaccineE.coliB-Haqetal,1995;()Heat-labiletoxinImmunogenicandTacketetal,EnterotoxigenicE.coliB-subunitTobacco,Potato,protectivewhen1998(human)MaizeadministeredorallyBArakawaVibriocholeraeCholeratoxinPotatoImmunogenicandetal,1998(human)B-subunitprotectivewhenadministeredorallyKapustaHepatitisBvirusEnvelopesurfaceImmunogenicwhenetal,1999;(human)proteinPotato,Lupin,administeredorallyRichterLettuceetal,2000NorwalkMasonNorwalkvirus(human)CapsidproteinTobacco,PotatoImmunogenicwhenetal,1996administeredorally_____McGarveyRabiesvirus(human)GlycoproteinTomatoetal,1995VP1CarrilloFoot-and-mouthdiseaseVP1structuralArabidopsis,Immunogenicandetal,1998;virus(agriculturalproteinAlfalfaprotectivewhenadminis-Wigdorovitzdomesticanimals)teredbyinjectionororallyetal,1999STubolyetal,GastroenteritisGlycoproteinS2000coronavirus(pig)Arabidopsis,ImmunogenicwhenTobacco,Maizeadministeredbyinjectionororally(Schillbergetal,2001)3.1immunomodulationantisensecosuppression(DeJaegeretal,2000)(1)(2)(3)(4)(5)3.2(intracellularimmunization)(extracellularimmunizationRNAZimmermann1998TMVscFvTMVscFv3Table3ExpressionlevelsofselectedbiopharmaceuticalsintransgenicplantsProteinsexpressedPotentialapplicationExpressionExpressionlevelsReferencessystemC()0.01%TSPCrameretal,HumanproteinCAnticoagulantTobacco1996(serumprotease)0.3%Crameretal,HumanhirudinThrombininhibitorBrassicanapus0.30%seedprotein19990.01%TSPMatsumotoHumanerythropoietinAnemiatreatmentTobaccoetal,19950.01%TSPStaubetal,HumansomatotropinGrowthhormoneTobacco20000.01%TSPHigoetal,HumanepidermalgrowthWoundrepairTobacco1993-0.01%EldelbaumHumaninterferonHepatitisCandBTobacco0.01%freshetal,1992treatmentweight0.02%TSPKusnadiHumanserumalbuminLivercirrhosisTobaccoetal,1997treatment0.05%DieryckHumanhemoglobin,BloodsubstituteTobacco0.05%seedproteinetal,199790%IgG5FabsscFv(DeJaegeretal,2000)3.3(Wilson,1993;Baulcombe,1994)1993(Tavladorakietal,1993)scFvTMVTMV(Vossetal,1995)0.23%TSP70%scFv(Zimmermannetal,1998)(Lomonossoff,1995)RNA(Borjaetal,1999;Rubioetal,1999)3.4(Mourguesetal,1998),LeGall1998phytoplasmascFvphytoplasmascFvphytoplasma3.5root-knotMeloidogyneincognita(Smantetal,1998;Rossoetal,1999)(Schillbergetal,2001)(Yuanetal,1998)(Yuanetal,1997)4(DeJaeger