Isolation and characterization of an anther-specific gene, RA8, from rice (Oryza sativa L.)
Isolation and characterization of an anther-specific gene, RA8, from rice (Oryza sativa L.)
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DOI：
10.1023/a:1006157603096
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发表时间：
1999-01-01
影响因子：
5.1
通讯作者：
An, GH
中科院分区：
文献类型：
--
作者：
Jeon, JS;Chung, YY;An, GH
通过差异筛选，从水稻花药文库中分离到水稻花药特异性cDNA克隆RA8。RNA印迹分析表明，RA8转录物特异性存在于花药中，随着花的成熟，转录物水平升高，在成熟花中达到最高水平。RA8克隆包含264个氨基酸残基的开放阅读框，并带有疏水n端区。推导出的氨基酸序列与已知序列没有明显的同源性。基因组DNA印迹分析显示RA8为单拷贝基因。分离到一个与RA8 cDNA相对应的基因组克隆，并将其启动子区与β -葡萄糖醛酸酶（GUS）基因融合。转基因水稻表现出GUS报告基因的花药特异性表达。组织化学GUS分析表明，RA8启动子在花药绒毡层、绒毡层和结缔组织中均有活性。实验表明，该基因的表达始于小孢子从四分体中释放出来，在液泡化花粉后期达到最高水平。RA8启动子可用于控制谷类植物花药中的基因表达，并可用于雄性不育植株的产生。
An anther-specific cDNA clone of rice, RA8, was isolated from an anther cDNA library by differential screening. RNA blot analysis indicated that the RA8 transcript is present specifically in anthers and the transcript level increased as flowers matured, reaching the highest level in mature flowers. The RA8 clone contains an open reading frame of 264 amino acid residues with a hydrophobic N-terminal region. The deduced amino acid sequences did not show significant homology to any known sequences. Genomic DNA blot analysis showed that RA8 is a single-copy gene. A genomic clone corresponding to the RA8 cDNA was isolated and its promoter region was fused to the beta-glucuronidase (GUS) gene. Transgenic rice plants exhibited anther-specific expression of the GUS reporter gene. Histochemical GUS analysis showed that the RA8 promoter was active in the tapetum, endothecium, and connective tissues of anthers. Experiments showed that expression of the gene starts when microspores are released from tetrads, and it reaches to the maximum level at the late vacuolated-pollen stage. The RA8 promoter may be useful for controlling gene expression in anthers of cereal plants and for generating male-sterile plants.