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1、福建林學(xué)院學(xué)報(bào)2014,34(2):152—157第34卷第2期JournalofFujianCollegeofForestry2014年4月葉子花花色DOD基因全長(zhǎng)cDNA克隆與序列分析柏忠良,佘文琴,潘東明,郭懷攀(1.福建農(nóng)林大學(xué)園藝學(xué)院,福建福州350002;2.福建農(nóng)林大學(xué)園藝產(chǎn)品采后貯藏與保鮮研究所,福建福州350002)摘要:以葉子花花苞為材料,運(yùn)用RACE技術(shù)克隆出了葉子花苞片中的多巴雙加氧酶基因cDNA全長(zhǎng),探討葉子花花色變化機(jī)理和甜菜色素代謝途徑。該序列長(zhǎng)度為1059bp,開(kāi)放閱讀框位于56—802bp之間,共
2、編碼249個(gè)氨基酸殘基。對(duì)該氨基酸序列進(jìn)行生物信息學(xué)分析,結(jié)果表明該氨基酸序列具有多巴雙加氧酶保守結(jié)構(gòu)域,所組成的蛋白分子量為27.94kD,等電點(diǎn)為5.48,是穩(wěn)定蛋白。對(duì)其二級(jí)結(jié)構(gòu)進(jìn)行預(yù)測(cè):該序列具有8個(gè)0【螺旋、8個(gè)B轉(zhuǎn)角、20個(gè)B折疊,其序列大多是由親水性氨基酸殘基組成,不具有跨膜結(jié)構(gòu)和信號(hào)肽。對(duì)該氨基酸序列進(jìn)一步的Blast比對(duì),發(fā)現(xiàn)組成多巴雙加氧酶的氨基酸序列因植物種類(lèi)而異,且親緣關(guān)系越遠(yuǎn),差異越大,結(jié)合花青素與甜菜色素代謝途徑的不同和在自然界中不能共存的現(xiàn)象,表明多巴雙加氧酶基因在生成甜菜色素植物中具有獨(dú)特的保守性。
3、關(guān)鍵詞:葉子花;多巴雙加氧酶;甜菜色素;基因克隆中圖分類(lèi)號(hào):Q781文獻(xiàn)標(biāo)識(shí)碼:A文章編號(hào):1001—389X(2014)02—0152—06Cloningandsequenceanalysisoffull-lengtheDNAoftheBougainvilleaspectabifisWilld.flowercolorDoDgeneBAIZhong—liang,,SHEWen—qing’,PANDong.ming',GU0Huai.pan,(1.CollegeofHorticulture,F(xiàn)uiianAgricuhureandFo
4、restryUniversity,F(xiàn)uzhou,F(xiàn)ujian350002,China;2.InstituteofPostharvestScienceandTechnologyofHorticulturalProducts,F(xiàn)ujianagricultureandForestryUniversity,F(xiàn)uzhou,F(xiàn)ujian350002,China)Abstract:TheRNAofBougainvilleaspectabilisWilld.bractswereextracted,andthe4,5-DOPAdioxygenase
5、(DOD)genefull—lengtheDNAwasclonedbyRACEtechnique.ThefulllengthofthecDNAwas1059bp,openreadingflame(ORF)waslocatedbetweenthe56—802bp,whichencoding249aminoacidresidues.Accordingtobioinformatieanalyticalresultsoftheaminoacidsequence,therewas4,5-DOPAdioxygenaseconserveddom
6、aininthissequence,andthemolecularweightoftheproteincon—ponentwas27.94kD.Thep1was5.48,anditwasstableprotein.Thepredictionofthesecondarystructureshowedthattherewere8d—coils,8B—sheets,and20p—turnsinit.Mostoftheaminoacidresidueswerehydrophilic,yetnosignalpeptideandtransme
7、m—branestructurewasfound.Theblastresultsshowedthatasthedifferencesofthespeciesincreased,thesimilarityoftheDODwasdecreased.ItisindicatedthatDODgenewasquiteconservedinbetalainplantsbydiscoveringthephenomenonthatanthocyaninsmetabolismandbetalainmetabolismweredifferentand
8、couldnotcoexistinoneplant.Keywords:BougainvilleaspectabilisWilld.;4,5-DOPAdioxygenase;betalain;genecloning葉子花(Bougainvilleas