Chen Yu, Qu Le-Qing, Jia Xu
State Key Laboratory of Plant Cell & Chromosome Engineering, Institute of Genetics and Development Biology, Chinese Academy of Sciences, Beijing 100101, China.
Yi Chuan Xue Bao. 2004 Mar;31(3):281-6.
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.
为了研究水稻谷蛋白胚乳特异性启动子在中国品种中花8号(Oryza sativa L. subsp japonica)中的表达模式,将UidA基因分别与水稻谷蛋白GluA-2基因5'上游2.3 kb和750 bp的序列融合,并通过农杆菌介导转化法导入水稻。Southern杂交表明UidA基因以单拷贝形式整合到转基因植物基因组中。Northern杂交显示UidA基因和内源GluA-2基因的表达分别在授粉后13 - 15天和11 - 13天达到最高水平,随后下降。对未成熟转基因水稻种子的组织化学染色表明UidA基因在胚乳中特异性表达,并且在糊粉层中观察到最高水平的GUS表达。GUS活性的定量分析表明,2.3 kb转基因植株种子的GUS活性约为750 bp转基因植株种子的两到三倍。序列分析表明位于 - 2,170 bp(从转录起始位点起)的G - box可能是一个定量顺式元件。