大豆GmbZIP33基因启动子的克隆及瞬时表达分析  

Cloning of GmbZIP33 Promoter from Soybean and Its Transient Expression Analysis in Arabidopsis thaliana

作　　者：白丽娟 刘薇 王志莉 王文婷 吴存祥[2] 冯永君[1] BAI Li-juan;LIU Wei;WANG Zhi-li;WANG Wen-ting;WU Cun-xiang;FENG Yong-jun(School of Life Science, Beijing Institute of Technology, Beijing 100081 , China;Institute of Crop Science, Chinese Academy of Agricultural Science,Beijing 100081 , China;Crop Institute, Shandong Academy of Agricultural Sciences, Jinan 250100, China;School of Life Science,The ChineseUniversity of Hong Kong, Hong Kong 999077 , China)

机构地区：[1]北京理工大学生命学院,北京100081 [2]中国农业科学院作物科学研究所,北京100081 [3]山东省农业科学院作物研究所,山东济南250100 [4]香港中文大学生命科学院,香港999077

出　　处：《大豆科学》2019年第4期511-516,共6页Soybean Science

基　　金：国家自然科学基金(31271636,31870090)

摘　　要：启动子作为基因调控的重要元件,决定着下游基因表达的时空和强度特性。为研究大豆GmbZIP33基因启动子功能,在前期克隆获得GmbZIP33基因(Glyma.03G219300.1)序列的基础上,通过N-PCR(nested PCR)技术克隆了GmbZIP33 CDS上游启动子区序列(2 316 bp)。利用PlantCARE在线分析软件进行顺式作用元件的预测,发现该序列上除含有TATA-box和CAAT-box等核心调控元件外,同时包括与抗逆、光响应、激素响应、蔗糖应答元件和多个与组织特异性表达相关的元件。为研究GmbZIP33启动子的表达调控特性,构建了该启动子调控报告基因GUS表达的载体,并利用农杆菌介导的花序浸染法将其转入拟南芥中,发现该启动子驱动下的GUS基因在不同组织(莲座叶、花、荚果和根)的维管束中均特异性表达;对转基因拟南芥进行实时荧光定量PCR检测发现,GUS基因在花中表达量最高,荚果中次之,表明GmbZIP33基因可能受到多种因素和蔗糖的调节并参与花荚发育的调控。Promoter, as an important element in regulating gene expression, determines the temporal, spatial and intensity characteristics of the downstream gene expression. To study the promoter function of soybean GmbZIP33 gene, we cloned 2 316 bp upstream region of GmbZIP33 CDS as the promoter by N-PCR(nested PCR) technique on the basis of previous cloned GmbZIP33 gene sequence, and predicted its cis-acting elements by using PlantCARE online analysis software. It was found that besides the core regulatory elements of TATA-box and CAAT-box, there were also stress resistance response, light response, hormone response, sucrose response elements and tissue-specific expression elements. A vector to regulate the expression of reporter gene GUS was constructed and transformed into Arabidopsis thaliana by Agrobacterium-mediated floral-dip method to study the expression characteristics of GmbZIP33 promoter. Histochemical staining of transgenic Arabidopsis showed that GUS gene driven by this promoter was specifically expressed in vascular bundles of different tissues(rosette leaf, flower, pod and root). Real-time fluorescence quantitative PCR of the GUS gene in these four tissues of transgenic Arabidopsis showed that it was most expressed in flower, followed by in pod, suggesting that GmbZIP33 gene might be regulated by many factors and sucrose, and showing the possibility of roles in flower and pod development regulation.

关 键 词：大豆 拟南芥 GUS 维管组织特异性启动子 

分 类 号：Q943.2[生物学—植物学] S565.1[农业科学—作物学]