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首页> 外文期刊>In Vitro Cellular and Development Biology. Plant: Journal of the Tissue Culture Association >Development and characterization of a chimaeric tissue-specific promoter in wheat and rice endosperm
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Development and characterization of a chimaeric tissue-specific promoter in wheat and rice endosperm

机译：小麦和水稻胚乳中嵌合组织特异性启动子的开发与表征

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The recently achieved significant improvement of cereal transformation protocols provides facilities to alter the protein composition of the endosperm, for example, to increase or decrease the quantity of one of its protein components or to express foreign molecules. To achieve this goal, strong endosperm-specific promoters have to be available. The aim of our work was to develop a more efficient tissue-specific promoter which is currently used. A chimaeric promoter was assembled using the 5′ UTR (1,900 bp) of the gene coding for the 1Bx17 HMW glutenin subunit protein, responsible for tissue-specific expression and the first intron of the rice actin gene (act1). The sequence around of the translation initial codon was optimized. The effect of the intron and promoter regulatory sequences, using different lengths of 1Bx17 HMW-GS promoter, were studied on the expression of uidA gene. The function of promoter elements, promoter length, and the first intron of the rice actin gene were tested by a transient expression assay in immature wheat endosperm and in stable transgenic rice plants. Results showed that insertion of the rice act1 first intron increased GUS expression by four times in transient assay. The shortest 1Bx17 HMW-GS promoter fragment (173 bp) linked to the intron and GUS reporter gene provided almost the same expression level than the intronless long 1Bx17 HMW-GS promoter. Analysis of the stable transformant plants revealed that 173 nucleotides were sufficient for endosperm-specific expression of the uidA gene, despite 13 nucleotides missing from the HMW enhancer sequence, a relevant regulatory element in the promoter region.
机译：最近实现的谷物转化方案的重大改进提供了改变胚乳蛋白质组成的设施，例如增加或减少其蛋白质成分之一的数量或表达外来分子的设施。为了实现这个目标，必须有强大的胚乳特异性启动子。我们工作的目的是开发当前使用的更有效的组织特异性启动子。使用编码1Bx17 HMW谷蛋白亚基蛋白的基因的5'UTR（1,900 bp）组装了嵌合启动子，该基因负责组织特异性表达和水稻肌动蛋白基因的第一个内含子（act1）。优化了翻译起始密码子周围的序列。研究了使用不同长度的1Bx17 HMW-GS启动子的内含子和启动子调控序列对uidA基因表达的影响。水稻肌动蛋白基因的启动子元件，启动子长度和第一个内含子的功能通过瞬时表达试验在未成熟的小麦胚乳和稳定的转基因水稻植株中进行了测试。结果显示，在瞬时分析中，插入水稻act1第一内含子可使GUS表达增加四倍。与内含子和GUS报告基因连接的最短1Bx17 HMW-GS启动子片段（173 bp）提供的表达水平与无内含子长1Bx17 HMW-GS启动子几乎相同。对稳定的转化植物的分析显示，尽管HMW增强子序列（启动子区域中的相关调控元件）缺失了13个核苷酸，但173个核苷酸足以用于胚乳特异性表达uidA基因。

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