黄花蒿AaCYP71AV1基因的克隆及植物表达载体的构建

黄花蒿AaCYP71AV1基因的克隆及植物表达载体的构建CLONINGOFAaCYP71AV1GENEANDCONSTRUCTIONEXPRESSIONVECTORFROMARTEMISIAANNUAl目录1前言 [39]。综上，我们给出了一种新型的提高青蒿素合成量的调控策略：将基于青蒿素生物合成途径的次生代谢调控和基于青蒿素合成部位的腺毛特征调控结合起来，由此来共同提高青蒿素的产量。目前，社会上对转基因技术存在一定的疑虑与偏见，但我认为：随着科学技术对我们日常生活的不断渗入，互联网时代下人们的信息交流愈加透明，相关制度不断地建立和完善，在不远的将来，人们会看到成熟的植物基因工程技术的伟大之处，尤其是在生产青蒿素等天然抗疟疾药物的过程中。参考文献屠呦呦.青蒿及青蒿素类药物[M].北京：化学工业出版社:北京市东城区青年湖南街13号,2009:1-260.赵燏黄.祁州药志.第一集.国立北平研究院，1936.58.中国中医研究院中药研究所.青蒿抗疟研究专辑1971～1978.中药研究资料,1978,3.青蒿素结构协作组.一种新型的倍半萜内酯——青蒿素[J].科学通报,1977,3:142.屠呦呦,倪慕云,钟裕蓉等.中药青蒿化学成分的研究[I].药学学报,1981,16(5):366-370.屠呦呦,倪慕云,钟裕蓉等.中药青蒿的化学成分和青蒿素衍生物的研究（简报）[J].中药通报,1981,6(2):31.ChinacooperativeresearchgrouponQinghaosuanditsderivativesasantimalarials.ChemicalstudiesonQinghaosu(Artemisinine)[J].JournalofTraditionalChineseMedicine,1982,2(1):3-8.TuYY,NiMY,ZhongYR,etc.StudiesontheconstituentsofArtemisiaannua:partⅡ[J].PlantaMedica,1982,44(3):143-145.屠呦呦,尹建平,吉力等.中药青蒿化学成分的研究（Ⅲ）[J].中草药,1985,16(5):200-201.屠呦呦,朱启聪,沈星.中药青蒿幼株的化学成分研究[J].中药通报,1985,10(9):419-420，402.KlaymanDL..Qinghaosu(artemisnin):anantimalarialdrugfromChina.[J].Science,1985,228:1049-1055.ZhaoX，LiuM，LiJ，etal．AntimalarialagentartesunateprotectsconcanavalinA－inducedautoimmunehepatitisinmicebyinhibitinginflammatoryresponses［J］．ChemBiolInteract，2017，274:116．TranTH，NguyenTD，PoudelBK，etal．Developmentandevaluationofartesunate-loadedchitosan-coatedlipidnanocapsuleasapotentialdrugdeliverysystemagainstbreastcancer［J］.AAPSPharmSciTech，2015，16(6):1307.ShiC，LiH，YangY，etal．Anti－inflammatoryandimmunoregulatoryfunctionsofartemisininanditsderivatives［J］．MediatorsInflamm，2015，doi:10.1155/2015/435713．WallAartTE,ParsN,BeekmanAC,etc.SeasonalvariationofatemisinanditsbiosynthicprecursorsinplantsArtemisiaannuaofdifferentgeographicalorigin:proofforexistenceofchemotypes[J].PlantaMed,1999,66:57-62.MaDM，WangZ，WangL，etal．Agenome－widescenarioofterpenepathwaysinself-pollinatedArtemisiaannua［J］．MolPlant，2015，8(11):1580．RamM，KhanM，JhaP，etal．HMG－CoAreductaselimitsartemisininbiosynthesisandaccumulationinArtemisiaannuaL．plants［J］．ActaPhysiolPlant，2010，32(5):859SchramekN，WangH，RMisch-marglW，etal．ArtemisininbiosynthesisingrowingplantsofArtemisiaannua．A13CO2study［J］．Phytochemistry，2010，71(2/3):179．AbdinMZ，AlamP．Geneticengineeringofartemisininbiosynthesis:prospectstoimproveitsproduction［J］．ActaPhysiolPlant，2015，37(2):33．NguyenKT，ArsenaultPR，WeathersPJ．Trichomes+roots+ROS=artemisinin:regulatingartemisininbiosynthesisinArtemisiaannuaL［J］．InVitroCellDevBiolPlant，2011，47(3):329．WangH，HanJ，KanagarajanS，etal．Trichome－specificexpressionoftheamorpha-4，11-diene12-hydroxylase(cyp71av1)gene，encodingakeyenzymeofartemisininbiosynthesisinArtemisiaannua，asreportedbyapromoter-GUSfusion［J］．PlantMolBiol，2013，81(1/2):119．RoDK，ParadiseEM，OuelletM，etal．Productionoftheantimalarialdrugprecursorartemisinicacidinengineeredyeast［J］．Nature，2006，440(7086):940．PorterTD,CoonMJ.CytochromeP450.Multiplicityofisoforms,substrates,calalyticandregulatorymechanisms[J].JBiolChem,1991,95:4126-4133.史超,邓洁,常振战.青蒿素生物合成关键酶CYP71AV1的表达纯化和晶体培养[J].中国中药杂志,2018,43(15):3064-3069.BerteaCM,FreijeJR,vanderWoudeH,etc.IdentificationofintermediatesandenzymesinvoledintheearlystepsofarteisininbiosynthesisinArtemisiaannua[J].PlantaMed,2005,71:40-47.KeatHT,DevinRP,DarwinWR,etc.ArtemisiaannuaL(Asteraceae)trichome-specificcDNAsrevealCYP71AV1,acytochromeP450withakeyroleinthebiosynthesisoftheantimalarialsesquiterpenelactoneartemisinin[J].FEBSlett,2006,580:1411-1416.TeohKH，PolichukDR，CovelloPS，etal．ArtemisiaannuaL．(Asteraceae)trichome-specificcDNAsrevealCYP71AV1，acytochromeP450withakeyroleinthebiosynthesisoftheantimalarialsesquiterpenelactoneartemisinin［J］．FEBSLett，2006，580(5):1411．王红.真菌诱导子对青蒿发根生长和青蒿素生物合成的影响及开花与青蒿素生物合成的相关性研究[D].北京:中国科学院植物研究所,2001.KomoriA，SuzukiM，MuranakaT，etal．ComparativefunctionalanalysisofCYP71AV1naturalvariantsrevealsanimportantresidueforthesuccessiveoxidationofamorpha-4，11-diene［J］．FEBSLett，2013，587(3):278．刘春朝.黄花蒿组织培养及培养物中青蒿素含量的研究[D].河南:河南大学,2007.陈大华,孟玉玲,叶和春等.青蒿转杜松烯合成酶基因发根系的培养[J].植物学报,1998,40:711-714.ChenDH,LiuCJ,YeHC,etc.Ri-mediatedtransformationofArtemisiaannuawitharecombinantfarnesyldiphosphatesynthasegeneforartemisininproduction[J].PlantCellTissOrgCult,1999,57:157-162.ChenDH,YeHC,LiGF.ExpressionofachimericfarnesyldiphosphateaynthasegeneinArtemisiaannuaL.transgenicplantsviaAgrobacteriumtumefaciens-mediatedtransfoemation[J].PlantSci,2000,155:179-185.GengS,MaM,YeHC,etc.EffectsofiptgeneexpressiononthephysiologicalandchemicalcharacteristicsofArtemisiaannuaL.[J].PlantSci,2001,160:691-698.WangH,GeL,YeHC,etc.Studiesontheeffectsoffpf1geneonArtemisiaannuafloweringtimeandonthelinkagebetweenfloweringandartemisininbiosynthesis[J].PlantaMed,2004,70:347-352.HanJL,LiuBY,YeHC,etc.EffectsofoverexpressionofArtemisiaannuafarnesyldiphosphatesynthaseontheartemisnincontentinArtemisiaannua[J].JournalofIntegrativePlantBiology,2006,48:482-487.QianShen,LidaZhang,ZhihuaLiao.TheGenomeofArtemisiaannuaProvidesInsightintotheEvolutionofAsteraceaeFamilyandArtemisininBiosynthesis[J].MolecularPlant,2018,43(03):1-