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Cloning and expression analysis of 4-diphosphocytidyl-2-C-methyl-D- erythritol kinase gene in Cinnamomum camphora

樟树4-二磷酸胞苷-2-C-甲基-D-赤藓醇激酶基因的克隆及表达分析



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Cloningandexpressionanalysisof4diphosphocytidyl2CmethylD
erythritolkinasegeneinCinnamomumcamphora
JINGLi1,ZHENGHan1,2,YAONa1,CHENMeilan1,SHENYe1,HUANGLuqi1
(1.StateKeyLaboratoryofDaodiHerbs,NationalResourceCenterforChineseMateriaMedica,
ChinaAcademyofChineseMedicalSciences,Beijing100700,China;
2.AnHuiUniversityofChineseMedicine,Hefei230038,China)
[Abstract] The4diphosphocytidyl2CmethylDerythritolkinasewasthefourthkeyenzymesinplantterpenoidbiosynthesispath
wayofmethylerythritolphosphatepathway(MEP)AccordingtothestudyofCinnamomumcamphoratranscriptomedata,weabtained
the4diphosphocytidyl2CmethylDerythritolkinasegeneusingRTPCR,andnamedCcCMK1,thendepositeditinGeneBank(Acces
sionnumber:Ku376098)Bioinformaticsanalysisshowedtheopenreadingframe(ORF)oftheCcCMK1was1212bpTheputative
proteinencoded403aminoacids,anditsmolecularweightwas4446kDaandtheoreticalyisoelectricpointwas499Transmembrane
structureanalysisshowedthattherewasnotransmembranestructureSignalpeptideanalysisshowedthatitwasanonsecretoryprotein,
andtherewasnosignalpeptideThesubcelularlocalizationshowedthatthechloroplastwaslocatedinthechloroplastAnalysisofthe
expressionofCcCMK1geneinfivechemotypesofCcamphorausingRealtimePCRshoweditsexpressionlevelwashighestin
Clongepaniculatum,andthelowestinBorneolcamphorThisresearchprovidedabasisforcharacterizingthekeyenzymegenesofter
penoidbiosyntheticpathwayinCcamphora
[Keywords] Cinnamomumcamphora;4diphosphocytidyl2CmethylDerythritolkinase(CMK);genecloning;bioinformation
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