棉花光合基因GhRCAα启动子的克隆及活性分析

发布时间：2018-11-16 16:33

【摘要】：为了克隆棉花Rubisco活化酶基因(RCA)启动子,研究其表达调控的分子机制,以百棉1号为材料,对GhRCAα启动子区2 000 bp的片段进行克隆、顺式作用元件分析以及活性分析,结果表明,许多重要的顺式作用元件包括响应于光、生物钟、逆境胁迫、植物激素以及其他的基本顺式作用元件特异地存在于GhRCAα启动子区;进一步对GhRCAα进行表达特性分析发现,该基因在光合作用进行的主要位置叶片中表达量最高,在其他组织表达量很低,其表达具有组织特异性,这与该启动子区存在许多光响应及组织特异性表达相关元件的结果相一致;将克隆的GhRCAα启动子片段以烟草叶片为受体材料进行瞬时表达分析表明,GhRCAα启动子可以驱动GUS基因的表达,表明克隆的启动子片段具有驱动目标基因表达的活性。克隆的GhRCAα启动子可能是一种组织特异型启动子,有望用于植物的遗传转化,进而更好地调控重要基因的特异性表达。
[Abstract]:In order to clone the (RCA) promoter of cotton Rubisco activase gene and study the molecular mechanism of its expression regulation, the 2 000 bp fragment of GhRCA 伪 promoter was cloned, the cis acting element analysis and the activity analysis were carried out using Baimian 1 as the material. The results show that many important cis-acting elements, including the response to light, biological clock, stress, plant hormones and other basic cis-acting elements, are specifically present in the GhRCA 伪 promoter region. Further analysis of the expression characteristics of GhRCA 伪 showed that the expression of GhRCA 伪 was the highest in the main position of photosynthesis, but very low in other tissues, and its expression was tissue specific. This is consistent with the results of many light responses and tissue-specific expression of related elements in the promoter region. The transient expression analysis of the cloned GhRCA 伪 promoter using tobacco leaves as the receptor material showed that the GhRCA 伪 promoter could drive the expression of GUS gene, indicating that the cloned promoter fragment had the activity of driving the expression of the target gene. The cloned GhRCA 伪 promoter may be a tissue-specific promoter, which is expected to be used in the genetic transformation of plants and to regulate the specific expression of important genes.
【作者单位】：河南科技学院生命科技学院现代生物育种河南省协同创新中心;河南省农业科学院;
【基金】：国家自然科学基金项目(31601347) 河南省高等学校重点科研计划项目(15A210001) 河南科技学院大学生创新训练计划(2016CX006)
【分类号】：S562

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