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《甘藍(lán)型油菜2個(gè)gpat6同源基因的克隆與表達(dá)分析》由會(huì)員上傳分享，免費(fèi)在線閱讀，更多相關(guān)內(nèi)容在學(xué)術(shù)論文-天天文庫(kù)。
1、作物學(xué)報(bào)ACTAAGRONOMICASINICA2014,40(7):1304?1310http://zwxb.chinacrops.org/ISSN0496-3490;CODENTSHPA9E-mail:xbzw@chinajournal.net.cnDOI:10.3724/SP.J.1006.2014.01304甘藍(lán)型油菜2個(gè)GPAT6同源基因的克隆與表達(dá)分析11111,21,2,*劉聰肖旦望胡學(xué)芳鄔克彬官春云熊興華12湖南農(nóng)業(yè)大學(xué)作物基因工程湖南省重點(diǎn)實(shí)驗(yàn)室,湖南長(zhǎng)沙410128;湖南農(nóng)業(yè)大學(xué)油料作物研究所,湖南長(zhǎng)沙
2、410128摘要:甘油-3-磷酸?；D(zhuǎn)移酶(sn-glycerol-3-phosphateacyltransferase,GPAT)是三酰甘油生物合成的關(guān)鍵酶,催化三酰甘油合成的起始步驟,在植物中參與調(diào)節(jié)生長(zhǎng)發(fā)育、脂質(zhì)的合成和逆境應(yīng)答等過(guò)程。本試驗(yàn)通過(guò)RT-PCR從甘藍(lán)型油菜品種湘油15中克隆得到GPAT6基因的2個(gè)拷貝,分別命名為BnGPAT6-1(GenBank登錄號(hào)為KJ000117)和BnGPAT6-2(KC106728)。它們的編碼區(qū)序列(codingDNAsequence,CDS)均為1506bp,編碼501個(gè)
3、氨基酸。預(yù)測(cè)其蛋白結(jié)構(gòu)N端含有一個(gè)類鹵酸脫鹵酶(haloaciddehalogenase-likehydrolase,HAD-like)活性結(jié)構(gòu)域,C端含有一個(gè)溶血磷脂?；D(zhuǎn)移酶(lysophospholipidacyltransferase,LPLAT)功能域。蛋白序列比對(duì)和進(jìn)化樹(shù)分析表明,甘藍(lán)型油菜與白菜(B.rapa)、甘藍(lán)(B.oleracea)、擬南芥(Arabidopsisthaliana)和琴葉擬南芥(A.lyrata)中的GPAT6序列相似性最高。組織表達(dá)結(jié)果表明BnGPAT6基因在花中的表達(dá)量最高,在未成
4、熟的胚中表達(dá)量呈先升高后降低的趨勢(shì)。BnGPAT6的表達(dá)受ABA的抑制;在干旱和6-BA脅迫下幾乎同時(shí)升高;在鹽脅迫下,短時(shí)間內(nèi)升高,達(dá)到峰值后降低;在水漬條件下沒(méi)有明顯變化。關(guān)鍵詞:甘藍(lán)型油菜;甘油-3-磷酸?；D(zhuǎn)移酶;基因克隆;表達(dá)分析;逆境脅迫CloningandExpressionAnalysisofTwoHomologousGenesCodingsn-Glycerol-3-PhosphateAcyltransferase6inBrassicanapus11111,21,2,*LIUCong,XIAODan-Wan
5、g,HUXue-Fang,WUKe-Bin,GUANChun-Yun,andXIONGXing-Hua12CropGeneEngineeringKeyLaboratoryofHunanProvince,HunanAgriculturalUniversity,Changsha410128,China;OilseedCropsInstitute,HunanAgriculturalUniversity,Changsha410128,ChinaAbstract:Akeyenzymesn-glycerol-3-phosphateac
6、yltransferase(GPAT)catalyzestheinitialstepofTAGbiosyntheticpathway,andisinvolvedinmanyprocessesincludingplantgrowth,developmentandresponsetoabioticstresses.Inthisstudy,twoB.napusGPAT6homologswereclonedfromtheleafofthecultivarxiangyou15usingRT-PCR,anddesignatedasBn
7、GPAT6-1andBnGPAT6-2,respectively.ThecodingDNAsequences(CDS)oftwoBnGPAT6genesare1506bpinlength,encodingtwodiffer-entpolypeptidesof501aminoacidresidues.Theproteinswerepredictedtoconsistofahaloaciddehalogenase-likehydrolasedomainandalysophospholipidacyltransferasedom
8、ain.MultiplesequencealignmentsandphylogeneticanalysisofGPATproteinsshowedthatBnGPAT6-1andBnGPAT6-2sharehighsimilaritywithGPAT6proteinsfromB.rapa,B.olera
