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1、http://www.paper.edu.cnCloningandanalysisofacytochromeP450genefromriceZHUShi-Feng,CAOXue-Song,ZHONGYong-Wang,WEIChun-Hong,LIYi(Peking-YaleJointCenterforPlantMolecularGeneticsandAgrobiotechnology,NationalLaboratoryofProteinEngineeringandPlantGeneticEngineering,CollegeofLifeScience,Peking
2、University,Beijing,100871,China)E-mail:Firstauthor:zhushif@pku.edu.cnCorrespondingauthor:liyi@pku.edu.cnAbstract：CytochromeP450mono-oxygenases(P450s)areasuperfamilyofheme-dependentenzymeswhichplayimportantrolesinalmostalloforganisms.Basedonricegenomedatabase,ageneencodingaricecytochrome
3、P450mono-oxygenasewasclonedandsequenced(GenBankaccessionno.AY579214).Thecodingregionofthegenewas1530nucleotidesinlengthandencodingaproteinwith510aminoacidresiduesinlength.SequenceanalysesindicatedthatthecytochromeP450correspondstoanent-kaureneoxidase(KO)likeprotein,whichislikelytobeinvo
4、lvedinphytoanticipins/phytoalexinsbiosynthesis.Southernblotsanalysissuggestedthatthereareatleastfoursimilargenesencodingent-kaureneoxidaseorent-kaureneoxidaselikeproteinsinricegenome.Therefore,thegenereportedinthispaperwasnamedOsKOS1.RT-PCRanalysisshowedthatOsKOS1wasexpressedinvegetativ
5、eorgansandreproductiveorgans,butatahigherlevelinvegetativeorgans.TheexpressionlevelofOsKOS1inRDV(ricedwarfvirus)-infectedplantswasmuchlowerthanthatinhealthyplants,indicatingthatOsKOS1islikelytobeinvolvedindefendingRDV.Keywords：cytochromeP450,ent-kaureneoxidaselikeprotein,phytoanticipin/
6、phytoalexin,riceCytochromeP450mono-oxygenases(P450s)arealargegroupofheme-containingenzymesthatcatalyzeNADPH-andO2-dependenthydroxylationreactionsandparticipateinthemetabolismofvarietyofmolecules,includingendogenoussubstratesandforeigncompounds[1].P450s,whichcomposealargesuperfamily,have
7、beenfoundinbacteria,yeast,insects,fish,mammals,plants,andfungi[2].Inplants,cytochromeP450shaveprovedtobeinvolvedinthe1http://www.paper.edu.cnproductionofphenylpropanoids,alkaloids,terpenoids,lipids,cyanogenicglycosides,andglucosinolates,aswellasthebiosynthesisofplantgrowthregul
