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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



全 文 :2016
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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
·8751·
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doi:10.4268/cjcmm20160902
  
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·9751·
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Vol41,No.9 May,2016
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3 
á 
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·1851·
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Vol41,No.9 May,2016
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32(11):186.
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[J].
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[4] 
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±_§mB„lmÒ
¡¢
[J].
ZW
,2014,36(1):58.
[5] ShiXueping,ZhangCuijie,LiuQinhong,etal.Denovocom
parativetranscriptomeanalysisprovidesnewinsightsintosucrose
inducedsomaticembryogenesisincamphortree(Cinnamomum
camphoraL.)[J].BMCGenomics,2016,17(1):26.
[6] 
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Actin
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[J].
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EF1a
•›„éœ
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[J].
9²È&(wk ,2015,35(5):122. [8] YangTao,LiJing,WangHaoxin,etal.Ageraniolsynthasegene fromCinnamomum tenuipilum[J].Phytochemistry,2005,66 (3):285. [9] NewmanJD,ChappelJ.Isoprenoidbiosynthesisinplants:car bonpartitioningwithinthecytoplasmicpathway[J].CritRev BiochenMolBiol,1999,34(2):95. [10] RohmerM,KnaniM,SimoninP,etal.Isoprenoidbiosynthesisin bacteria:anovelpathwayfortheearlystepsleadingtoisopentenyl diphosphate[J].BiochenJ,1993,295:517. [11] LauleO,FürholzA,ChangHS,etal.Crosstalkbetweency tosolicandplastidialpathwaysofisoprenoidbiosynthesisinAra bidopsisthaliana[J].ProcNatlAcadSciUSA,2003,100(11): 6866. [12]  ºG , ›_ò , (IJ , É . h 4(5′ ‘’“ )2C ” • D –—˜™šœ cDNA w՝ž‚—˜Ÿ ¡¢ [J]. ,k ,2008,43(12):1251. [13]  èKµ . ?@ MEP ¬­ôO•›wÕ cDNA ¡v]`Æ OÈɉŠ [D]. 23 : %²È&‰Šþ
,2012.
[14] 
Lœy



MB , É . ØD 4(5′ ‘’“ )2C ” • D –—˜™š•›œw՝žŸ ¡¢ [J]. %, -. ,2015,40(21):4165. ·3851· 2016 y 5 z À 41 ÁÀ 9 } Vol41,No.9 May,2016 [15]  NO , èP¸ , Q þ , É . ¿©4L”Ñ1‘™š NsyMK • ›žÖÆO¡¢ [J]. ‡¨-. ,2013(5):75. [16]  !R , SòF , ßU² . Lk”Ñ1‘™š PnMVK1 •›œ ž#©¨vС¢ [J]. ,k ,2012,47(8):1092. [17] LoisLM,RodrI′guezConcepcio′nM,GalegoF,etal.Carote noidbiosynthesisduringtomatofruitdevelopment:regulatoryrole of1deoxyDxylulose5phosphatesynthase[J].PlantJ,2000, 22(6):503. [18] CareteroPauletL,AhumadaI,CunileraN,etal.Expression andmolecularanalysisoftheArabidopsisDXRgeneencoding 1deoxyDxylulose5phosphatereductoisomerase,thefirstcom mitedenzymeofthe2CmethylDerythritol4phosphatepath way[J].PlantPhysiol,2002,129(4):1581. [19] QuerolJ,CamposN,ImperialS,etal.Functionalanalysisof theArabidopsisthalianaGCPEproteininvolvedinplastidisopre noidbiosynthesis[J].FEBSLet,2002,514(2/3):343. [20] ShanShan,ChenXuehui.CrystalizationandpreliminaryXray analysisof4diphosphocytidyl2CmethylDerythritolkinase (IspE)fromMycobacteriumtuberculosis[J].ActaCrystalogr SectFStructBiolCrystCommun,2011,67(7):821. [21]  “)a , TU , ¡VW , É . %§°¨²>‚\¥«
¡
[M].


ä9&(Öm÷
,1997:57.
[22] LütgenH,RohdichF,HerzS,etal.Biosynthesisofterpe
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ProcNatlAcadSciUSA,2000,97(3):1062.

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