柠条锦鸡儿CkNCED1基因启动子的克隆及表达分析

利用染色体步移(Genome walking)技术,克隆柠条锦鸡儿CkNCED1基因上游启动子序列。经顺式元件预测分析,该序列除基本的启动元件之外,还含有2个脱落酸诱导响应元件ABRE、此外还含有多个逆境相关的元件。将CkNCED1基因启动子区连接到pGWB533植物表达载体上,构建Promoter::GUS载体,GUS组织化学染色结果表明,CkNCED1基因在植物的叶、根、茎、花和角果的维管组织中均有表达,且在叶片中表达强度最高。以上研究确定了柠条锦鸡儿CkNCED1基因的表达部位和强度。进一步说明CkNCED1基因在ABA合成调控中发挥重要作用,为深入研究基因功能奠定基础。

Cloning and expression analysis of the promoter of Caragana korshinskii gene

A genomic walking technique was used to clone the promoter sequence of the CkNCED1 gene from Caragana korshinskii. The plant CARE cis-element analysis revealed that, besides the basic elements, the promoter sequence included two ABA responsive elements (ABRE) and many other stress-response elements. The PGWB533 expression vector containing this promoter sequence was constructed with the GUS reporter gene. Histochemical staining analysis showed that the GUS reporter gene was expressed in the tissues of leaves, root, stem, flower and silique, and was especially higher in leaves. These results identified the localization and expression of the CkNCED1 gene in vivo, which suggested that CkNCED1 gene plays an important role in biosynthesis regulation of ABA. The study lay a foundation for further analysis of the gene function.