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1、作物學(xué)報(bào)ACTAAGRONOMICASINICA2014,40(11):1925?1935http://zwxb.chinacrops.org/ISSN0496-3490;CODENTSHPA9E-mail:xbzw@chinajournal.net.cnDOI:10.3724/SP.J.1006.2014.01925兩種苧麻纖維素合酶基因cDNA序列的克隆及表達(dá)1,**2,**1111劉昱翔陳建榮彭彥黃妤趙燕黃麗華21,*郭清泉張學(xué)文12湖南農(nóng)業(yè)大學(xué)生物科學(xué)技術(shù)學(xué)院,湖南長沙410128;長沙學(xué)院生物工程與環(huán)境科學(xué)系,湖南長沙410003摘要:從苧麻轉(zhuǎn)錄組數(shù)據(jù)出
2、發(fā),利用Blast工具從中分析出與多種植物纖維素合酶高度相似的片段CL789和Unigene20360。根據(jù)片段信息設(shè)計(jì)特異性引物,從苧麻[Boehmerianivea(Linn.)Gaud.]栽培種湘苧3號中克隆纖維素合酶核心片段,并利用5'及3'RACE技術(shù)獲得2個片段的全長cDNA。兩者都具有典型的纖維素合酶特征結(jié)構(gòu)域,表明為2個苧麻纖維素合酶基因CesA的cDNA序列,分別命名為BnCesA2和BnCesA3。BnCesA2基因編碼區(qū)全長度3240bp,編碼1079氨基酸多肽;BnCesA3基因編碼區(qū)全長3120bp,編碼1039氨基酸多肽。對BnCesA
3、2和BnCesA3基因在湘苧1號、湘苧3號、湘潭大葉白和城步青麻苧麻品種木質(zhì)部和韌皮部熒光定量PCR分析顯示,2個基因在不同品種苧麻的木質(zhì)部及韌皮部都有表達(dá),但表達(dá)量存在著一定差異,整體而言BnCesA2具有更高的表達(dá)水平,其木質(zhì)部和韌皮部的表達(dá)都為BnCesA3的2~5倍。推測BnCesA2和BnCesA3均參與了苧麻細(xì)胞壁的次生合成。關(guān)鍵詞:苧麻;纖維素合酶基因;cDNA克隆;表達(dá)分析cDNACloningandExpressionofTwoCelluloseSynthaseGenesfromBoeh-merianivea1,**2,**1111LIUYu-X
4、iang,CHENJiang-Rong,PENGYan,HUANGYu,ZHAOYan,HUANGLi-Hua,GUO21,*Qing-Quan,andZHANGXue-Wen12CollegeofBioscienceandBiotechnology,HunanAgriculturalUniversity,Changsha410128,China;DepartmentofBiotechnologyandEnviron-mentalScience,ChangshaUniversity,Changsha410003,ChinaAbstract:Twopotential
5、lyhighhomologousfragmentsCL789andUnigene20360wereidentifiedasplantcellulosesynthasecharactersequencefromthetranscriptomedataweobtainedpreviouslybyBlastaligningandhomologousscreening.ThetwopairsofspecificprimerswerethendesignedbasedontheCL789andUnigene20360sequencesinformation.Theinter
6、mediatefragmentsoftwocellulosesynthasegenecDNAwereclonedfromramievarietyXiangzu3byRT-PCR.AndthewholecDNAwasclonedbyfollowed5'and3'RACE.ThefulllengthcDNAsweresequencedandtheirencodedputativeproteinswereidentifiedascellulosesynthasebytheconserveddomainanalysis.The‘setwocDNAsequenceswere
7、namedasBnCesA2andBnCesA3respec-tively.Thefull-lengthcodingsequenceofBnCesA2geneis3240bp,andencodesaputative1079aminoacids.Thecodingse-quenceofBnCesA3geneis3120bp,andcouldbetranslatedintoa1039aminoacidsprotein.WedesignedthespecificprimersbasedoncDNAsequencesofthetwogenesandtheirexpress
8、ionle
