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茉莉花 j sggp ps 基因的克隆及生物信息学与表达分析

花匠小妙招
2024-11-20 17:08

孙君,林浥,俞滢,等.茉莉花JsGGPPS基因的克隆及生物信息学与表达分析[J].福建农业学报,2016,31(4):350-355.SUNJ,LINY,YUY,etal.Cloning,MolecularCharacterization,andExpressionofJsGGPPsGenefromJasminumsambac[J].FujianJournalofAgriculturalSciences,2016,31(4):350-355.茉莉花JsGGPPS基因的克隆及生物信息学与表达分析孙君1,2,林浥2,俞滢2,陈静2,姚雪倩2,陈桂信2,叶乃兴2∗(1福建省农业科学院茶叶研究所,福建福安355015;2福建农林大学园艺学院/茶学福建省高等学校重点实验室,福建福州350002)

收稿日期:2016-02-09初稿;2016-03-30修改稿作者简介:孙君(1988-),女,研究实习员,研究方向:茶树栽培与资源利用(Email:huiseshuijing@qqcom)∗通讯作者:**兴(1963-),男,教授,研究方向:茶学与茉莉资源利用(Email:ynxtea@126com)基金项目:福建省自然科学基金(2016J01110);福州市农业局2014年福州茉莉花茶产业提升项目(20143)摘要:牻牛儿基牻牛儿基焦磷酸合酶(Geranylgeranypyrophosphatesynthase,GGPPS)是萜烯类香气物质合成途径中的关键酶。采用RTPCR和RACE相结合的方法,克隆了茉莉花牻牛儿基牻牛儿基焦磷酸合酶基因的全长cDNA(命名为JsGGPPS,GenBank登录号AIY244211),采用实时荧光定量PCR技术分析JsGGPPS基因在茉莉花开放过程中的表达模式。结果表明,JsGGPPS全长cDNA为1410bp,含1083bp完整的开放阅读框(ORF),其推导的氨基酸序列包含360个氨基酸,属于transisoprenyldiphosphatesynthases(IPPS)和classIterpenecyclases家族,含链长控制区(CLDR)、2个天冬氨酸富集区以及其他保守区。JsGGPPS基因与独行菜Lepidiumapetalum、橡胶草Taraxacumkok-saghyz的同源性分别为91%、86%。实时荧光定量PCR结果表明,茉莉花开放过程中JsGGPPS基因在晚上18:00时表达量较低,呈上调趋势,在22:00时达到最高,呈下调趋势,在翌日2:00时表达量最低,与茉莉花释香趋势相似,为深入研究茉莉花萜烯类香气物质代谢途径奠定基础。关键词:茉莉花;萜烯类;香气;牻牛儿基牻牛儿基焦磷酸合酶;克隆;表达分析中图分类号:S6066文献标识码:A文章编号:1008-0384(2016)04-350-06Cloning,MolecularCharacterization,andExpressionofJsGGPPsGenefromJasminumsambacSUNJun1,2,LINYi2,YUYing1,CHENJing2,YAOXueqian2,CHENGuixin2,YENaixing2∗(1TeaResearchInstitute,FujianAcademyofAgriculturalSciences,Fu′an,Fujian355015,China;2CollegeofHorticulture,FujianAgricultureandForestryUniversity/KeyLaboratoryofTeaScienceatUniversitiesinFujian,Fuzhou,Fujian350002,China)Abstract:Geranylgeranypyrophosphatesynthase(GGPPS)catalyzesthebiosynthesisofgeranylgeranyldiphosphate,whichisakeyprecursorofthearomaticterpenesinjasmineflowers.ThefulllengthcDNAsequenceofGGPPSgeneintheterpenoidbackbonebiosynthesiswasclonedfromJasminumsambacbyRTPCRcombinedwithRACE,andsubsequentlynamedJsGGPPSwithaGenBankAccessionnumberofAIY244211TheexpressionofJsGGPPSduringjasminebloomingwasdetectedbyquantitativerealtimePCR.TheresultsshowedthatthefulllengthofthecDNAsequencewas1410bpwitha1083bpopenreadingframe.Thededucedproteinconsistedof360aminoacids,andshared91%identitywithGGPPSfromLepidiumapetalumand84%fromTaraxacumkok-saghyz.JsGGPPShadonechainlengthdeterminationregion,twohighlyconservedaspartaterichmotifs,andotherconserveddomains,whichbelongedtothetransisoprenyldiphosphatesynthasesandclassIterpenecyclasessuperfamily.ThequantitativerealtimePCRshowedthattheJsGGPPSexpressionat18:00waslow,andbegantoriseandreachedamaximumat20:00followedbyasteadydeclinetoaminimumat2:00.TheobservationsignificantlycorrelatedwiththearomareleasepatternofJsambacinnature.Theresultsprovidedabasisforfurtherstudyontheterpenebiosyntheticpathwayoftheflower.Keywords:Jasmimunsambac;terpene;fragrance;geranylgeranypyrophosphatesynthase;cloning;expressionanalysis.福建农业学报31(4):350~355,2016http://www.fjnyxb.cnFujianJournalofAgriculturalSciencesdoi:1019303/j.issn.10080384.2016.04.004