棉花种子特异表达α球蛋白A基因启动子的功能分析  

Functional Analysis of the Cotton Seed-Specific Expression of α-Globin A Gene Promoter(Gossypium hirsutum L.)

作　　者：王芳[1,2] 魏琦超[2] 张锐[2] 孙国清[2] 陈全家[1] 石书兵[1] 曲延英[1] 郭三堆[2] 

机构地区：[1]新疆农业大学农学院,乌鲁木齐830052 [2]中国农业科学院生物技术研究所,北京100081

出　　处：《棉花学报》2016年第3期189-198,共10页Cotton Science

基　　金：国家转基因重大专项:转基因耐旱耐盐碱棉花新品种培育(2014ZX08005-004)

摘　　要：研究植物种子特异启动子具有重要的理论和实际意义。本文研究了棉花α球蛋白A基因启动子,该启动子序列全长为1640 bp,作用元件分析表明该区域除了具有核心调控序列外,还含有多个与组织特异性相关的顺式作用元件。设计其5'端构建4个不同长度的缺失、融合GUS基因的表达载体,并通过蘸花法分别转化拟南芥。转基因拟南芥GUS表达分析结果表明,该启动子能驱动GUS基因在胚、露白的种子、子叶期的幼苗中表达,而二叶期的幼苗、根、茎、莲座叶、茎生叶和花苞组织则没有表达,说明棉花α球蛋白A基因启动子是一个种子特异性启动子。208 bp长度的启动子足以维持其种子特异表达功能,而且在启动子的-684和-208区域之间可能存在负调控元件或负调控区域。分析棉花α球蛋白A基因启动子是一个种子特异性启动子,其基本启动子区域不长于208 bp。The study of plant seed-specific promoters has important theoretical and practical significance. The α-globin A gene promoter sequence length is 1640 bp, and cis-acting element analysis revealed the region not only has the core control sequence,but also contains multiple function components associated with tissue specificity of cis-acting elements. Four fusions of GUS gene expression vectors were constructed using 5′ end deletions of different lengths. These were then transformed into plants using the Arabidopsis floral dipping method. Transgenic Arabidopsis GUS expression analyses demonstrated the promoter can drive the GUS gene to express in the embryos, testa split seeds, and seedlings at the cotyledon stage; while expression was not observed in the seedlings at the second leaf stage, roots, stems, rosette leaves, stem leaves, or buds. These indicate the cottonα-globin A gene promoter is seed specified. With a length of 208 bp, a promoter is sufficient to maintain seed specific expression function; there may be a negative regulatory element or negative regulatory region located in the-684 to-208 region of the promoter. Thus the cotton α-globin A gene promoter is seed-specific and its basic promoter region is at most 208 bp.

关 键 词：棉花 种子特异性启动子 α球蛋白A基因 功能分析 

分 类 号：S562[农业科学—作物学]