CAJ | 학술논문

为了研究谷蛋白胚乳特异性表达启动子在我国栽培稻品种中的表达模式,将UidA基因分别置于水稻谷蛋白GluA-2基因750 bp和2.3 kb上游序列下游,利用农杆菌转化法导入栽培稻品种中花8号并获得转基因植株.Southern blot检测表明,UidA基因已经整合到水稻基因组当中并以单拷贝存在.Northern blot检测表明,开花后13～15 d和11～13 d,UidA基因和水稻内源的GluA-2基因的表达量分别达到最高,随后逐渐降低.对转基因植株种子的GUS染色表明,UidA基因仅在胚乳中表达,在糊粉层中GUS表达量最高.测定了2.3 kb和750 bp转基因植株种子的GUS活性,结果表明前者的GUS活性是后者的2～3倍.序列分析表明,位于GluA-2基因转录启始位点上游2 170 bp的G-box可能是一个与表达量相关的顺式调控元件.
위료연구곡단백배유특이성표체계동자재아국재배도품충중적표체모식,장UidA기인분별치우수도곡단백GluA-2기인750 bp화2.3 kb상유서렬하유,이용농간균전화법도입재배도품충중화8호병획득전기인식주.Southern blot검측표명,UidA기인이경정합도수도기인조당중병이단고패존재.Northern blot검측표명,개화후13～15 d화11～13 d,UidA기인화수도내원적GluA-2기인적표체량분별체도최고,수후축점강저.대전기인식주충자적GUS염색표명,UidA기인부재배유중표체,재호분층중GUS표체량최고.측정료2.3 kb화750 bp전기인식주충자적GUS활성,결과표명전자적GUS활성시후자적2～3배.서렬분석표명,위우GluA-2기인전록계시위점상유2 170 bp적G-box가능시일개여표체량상관적순식조공원건.
In order to study the expression pattern of rice glutelin endosperm specific promoter in Chinese cultivar Zhonghua 8 (Oryza sativa L.subsp japonica),UidA gene was fused with rice glutelin GluA-2 gene 5′upstream sequence 2.3 kb and 750 bp upstream respectively and transferred into rice by Agrobacterium mediated transformation.Southern blot indicated that UidA gene was integrated into the genome of transgenic plants as single copy.Northern blot demonstrated that the expression of UidA gene and endogenous GluA-2 gene reached their highest level at 13～15 days and 11～13 days after pollination respectively,and then declined.Histochemical staining of immature transgenic rice seeds showed UidA gene was specifically expressed in endosperm and the highest level GUS expression was observed in aleurone layer.Quantitative analysis of GUS activity showed seeds GUS activity of that 2.3 kb transgenic plant was about two to three folds of those of 750 bp transgenic plant.Sequence analysis suggested that the G-box located in the -2 170 bp (from transcription start site) may be a quantitative cis-element.