大豆硬脂酸-ACP脱饱和酶基因启动子的克隆及其表达活性分析  

Cloning and Activity Analysis of Soybean SACPD-C Promoter

作　　者：张庆林[1] 赵艳[2] 李晓薇[1] 翟莹[1] 张艳[1] 王英[1] 李景文[1] 王庆钰[1] 

机构地区：[1]吉林大学植物科学学院,吉林长春130062 [2]齐齐哈尔大学生命科学与农林学院,黑龙江齐齐哈尔161006

出　　处：《作物学报》2011年第7期1205-1211,共7页Acta Agronomica Sinica

基　　金：国家转基因生物新品种培育重大专项(2008ZX08004-003);国家自然科学基金项目(30971808);吉林省科技发展计划重点项目(20080204);长春市科技局国际科技合作项目(08GH10);"211"三期建设项目资助

摘　　要：以吉豆2号基因组为模板,通过TAILPCR方法,扩增得到大豆硬脂酸-ACP脱饱和酶基因启动子片段SACPD-Cp。PLACE在线启动子预测分析表明,该序列中含有多种典型的种子特异性表达序列元件。将SACPD-Cp片段取代pCAMBIA1301质粒中的CaMV35S启动子,构建表达载体pCAM-SACPD-Cp,通过农杆菌介导法在大豆组织中进行瞬时表达,GUS组织化学染色和荧光定量研究其表达特性。结果表明,SACPD-Cp驱动GUS基因在种子中的表达活性是CaMV35S启动子的93.01%;SACPD-Cp启动子与现已知启动子无同源性,仅在大豆种子中检测到GUS活性,而在根、茎和叶组织中均未检测到GUS活性,证实SACPD-Cp是一个新的种子特异性启动子。The SACPD-Cp promoter of soybean SACPD-C was isolated from the genomic DNA of soybean cultivar Jidou 2 using TAIL PCR.Promoter sequence analysis by PLACE showed that the cloned fragment contained many motifs that constituted the seed-specific cis-elements.Replacing CaMV35S promoter of pCAMBIA1301 with the SACPD-Cp fragment,the binary expression vector pCAM-SACPD-Cp was constructed.Transient expression by Agrobacterium tumefaciens mediated method,the histochemical GUS analysis and fluorometric GUS analysis were used for testing the expression of the GUS activity.The results indicated that GUS activity driven by SACPD-Cp fragment was 93.01% of that driven by CaMV35S promoter.The SACPD-Cp promoter did not have the homology compared with the reported promoters.GUS activity assays indicated that GUS was expressed only in seeds,but not in roots,stems and leaves,which suggests the SACPD-Cp is a seed-specific promoter.

关 键 词：大豆 硬脂酸-ACP脱饱和酶基因 序列分析 种子特异性启动子 瞬时表达 

分 类 号：S565.1[农业科学—作物学]