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机构地区:[1]中国科学院遗传与发育生物学研究所植物细胞与染色体工程国家重点实验室,北京100101
出 处:《Acta Genetica Sinica》2004年第3期281-286,共6页
摘 要: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 750bp和 2 3kb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。Southernblot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。Northernblot检测表明 ,开花后 13~15d和 11~ 13d ,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 3kb和 750bp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 2170bp的GIn 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.
关 键 词:水稻谷蛋白GluA—2基因 表达模式 G—box
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