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首页> 外文期刊>African Journal of Biotechnology >Cloning and functional analysis in transgenic tobacco of a tapetum-specific promoter from Arabidopsis
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Cloning and functional analysis in transgenic tobacco of a tapetum-specific promoter from Arabidopsis

机译：转基因烟草拟南芥中绒毡层特异性启动子的克隆与功能分析

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摘要

The 5’-flanking region of 1174 bp upstream of the translation start point (TSP) of a reported?Arabidopsis?anther-specific gene,?Anther7?gene (ATA7), which putatively encodes a protein related to lipid transfer protein, was cloned and functionally analyzed in transgenic tobacco after been fused with β-glucuronidase (GUS) gene reporter. Histochemical GUS staining of the transgenic plants showed that the cloned fragment did drive GUS expression exclusively in the anther, not in any other parts of floral organs, including pollens and nor in any vegetative tissue. Transverse section of the GUS-blue anthers disclosed that the blue cells were present uniquely in the tapetum of the anther. A series of 5’-deletion of cloned fragment indicated that a short segment of 179 bp upstream of the TSP (-155 bp upstream of the transcription start site) retained not only the promoter’s driving power, but also its tapetum-specificity.?Cis-acting element search in this short segment revealed the presence of numbers of organ- and tissue-specific motifs, including pollen-specific LAT52 and SLG13. These results indicated that the tapetum-specificity of?ATA7gene is mainly conferred by its promoter, and such a promoter, in particular, the core one should be useful both for identification of tapetum-involved genes and for biotechnological applications.
机译：克隆了一个已报道的拟南芥花药特异基因，即Anther7基因（ATA7）的翻译起始点（TSP）上游1174 bp的5'侧翼区，该基因推测编码与脂质转移蛋白相关的蛋白。与β-葡萄糖醛酸酶（GUS）基因报告基因融合后，在转基因烟草中进行功能分析。转基因植物的组织化学GUS染色表明，克隆的片段确实确实在花药中驱动GUS表达，而不是在花器官的任何其他部分（包括花粉）和任何营养组织中都驱动GUS表达。 GUS蓝花药的横切面显示，蓝色细胞独特地存在于花药的绒毡层中。一系列克隆片段的5'缺失表明，TSP上游179 bp的短片段（转录起始位点上游-155 bp）不仅保留了启动子的驱动力，而且还保留了其绒毡层特异性。在这短段中的功能分子搜索揭示了器官和组织特异性基序的存在，包括花粉特异性LAT52和SLG13。这些结果表明，βATA7基因的绒毡层特异性主要是由其启动子赋予的，这种启动子，特别是核心启动子，对于鉴定绒毡层相关基因和生物技术应用都应是有用的。

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