[Expression of the GUS fusion gene controlled by the tomato rbcS3A promoter in transgenic rice].

植物生理与分子生物学学报 Pub Date : 2007-06-01
Qiao-Quan Liu, Heng-Xiu Yu, Wen-Juan Zhang, Zhi-Yun Gong, Ming-Hong Gu
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Abstract

To use different types of promoters in transgenic rice research, the 1.1 kb 5'-upstream regulation region of one of the tomato (Solanum tuberosum L.) Rubisco small subunit gene, rbcS3A, was cloned and its sequences were confirmed by comparison with the known sequences in GenBank. The cloned rbcS3A promoter was fused to the 5'-upstream of GUS (beta-glucuronidase) coding region in a binary vector, and introduced into an elite japonica rice variety by Agrogacterium-mediated transformation. The integration of the GUS fusion gene into the genome of transgenic rice was confirmed by both PCR and Southern blot 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 stem, leaf blade and sheath than in other organs of transgenic rice plants, and showed highest in the stem, which implies that the tomato rbcS3A promoter can make tissue-specific, in particular in the stem, expression of foreign genes in transgenic rice. The results present here also demonstrate that light induction had no effect on the expression of the foreign gene when regulated by the tomato rbcS3A promoter in transgenic rice. Our results show that the cloned tomato rbcS3A promoter might be very useful for the expression of target genes in transgenic rice, with particularly high efficiency in stem tissues.

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[番茄rbcS3A启动子调控GUS融合基因在转基因水稻中的表达]。
为了在水稻转基因研究中使用不同类型的启动子,其中一种番茄(Solanum tuberosum L.)的1.1 kb 5'-上游调控区克隆了Rubisco小亚基基因rbcS3A,并将其序列与GenBank中已知序列进行比对。克隆的rbcS3A启动子在二元载体中融合到GUS (β -葡萄糖醛酸酶)编码区5′上游,并通过agrogacterium介导转化导入到粳稻优良品种中。通过PCR和Southern blot分析证实GUS融合基因已整合到转基因水稻基因组中。组织化学染色和GUS活性定量分析结果显示,GUS融合基因在转基因水稻茎、叶片和鞘中的表达量明显强于其他器官,且在茎中表达量最高,说明番茄rbcS3A启动子可以使外源基因在转基因水稻中表达具有组织特异性,特别是在茎中。本研究结果还表明,在番茄rbcS3A启动子调控下,光诱导对外源基因在转基因水稻中的表达没有影响。我们的研究结果表明,克隆的番茄rbcS3A启动子可能对转基因水稻中靶基因的表达非常有用,在茎组织中的表达效率特别高。
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