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[水稻rbcS启动子调控外源基因在转基因水稻中的特异性表达]

[Specific expression of the foreign gene regulated by the rice rbcS promoter in transgenic rice].

作者信息

Liu Qiao-Quan, Yu Heng-Xiu, Zhang Wen-Juan, Wang Hong-Mei, Gu Ming-Hong

机构信息

Key Laboratory of Plant Functional Genomics of Jiangsu Province, College of Bioscience and Biotechnology, Yangzhou University, Yangzhou 225009, China.

出版信息

Zhi Wu Sheng Li Yu Fen Zi Sheng Wu Xue Xue Bao. 2005 Jun;31(3):247-53.

Abstract

To use different types of promoters in transgenic rice research, the 5'-upstream regulation region of rice Rubisco small subunit gene (rbcS) was cloned from a Chinese cultivar Wuyunjing 8, and its sequences were confirmed by comparison with the known genome sequences of both japonica and indica rice. The cloned rbcS promoter was fused to the 5'-upstream of GUS (beta-glucuronidase) coding region in a binary vector, and introduced into rice by Agrogacterium-mediated transformation. The integration of the rbcS-GUS fusion gene in transgenic rice was confirmed by PCR analysis. The results of both histochemical staining and quantitative analysis of GUS activity showed that the expression level of GUS fusion gene was significantly stronger in leaf blade and sheath than in other organs of transgenic rice plants, and the GUS activity was restricted to the mesophyll cells of leaf tissue, which showed that the rice rbcS promoter could control not only the tissue- but also the cell-specific expression of foreign genes in transgenic rice. The present results also demonstrated that light induction had a significant effect on the enhancement of transgene's expression when regulated by the rice rbcS promoter in transgenic rice. Our results showed that the rice rbcS promoter might be very useful for the expression of target genes in transgenic rice, with particularly high efficiency in leaf tissues.

摘要

为了在转基因水稻研究中使用不同类型的启动子,从中国品种武运粳8号中克隆了水稻核酮糖-1,5-二磷酸羧化酶/加氧酶小亚基基因(rbcS)的5'上游调控区,并通过与粳稻和籼稻已知基因组序列进行比较来确认其序列。将克隆的rbcS启动子与二元载体中GUS(β-葡萄糖醛酸酶)编码区的5'上游融合,并通过农杆菌介导的转化导入水稻。通过PCR分析确认了rbcS-GUS融合基因在转基因水稻中的整合。组织化学染色和GUS活性定量分析结果均表明,GUS融合基因在转基因水稻植株的叶片和叶鞘中的表达水平明显强于其他器官,且GUS活性局限于叶组织的叶肉细胞,这表明水稻rbcS启动子不仅可以控制转基因水稻中外源基因的组织特异性表达,还可以控制细胞特异性表达。本研究结果还表明,在转基因水稻中,当受水稻rbcS启动子调控时,光诱导对转基因表达的增强具有显著影响。我们的结果表明,水稻rbcS启动子可能对转基因水稻中靶基因的表达非常有用,在叶片组织中具有特别高的效率。

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