地上部特异性启动子驱动番茄原系统素表达的载体构建及转基因植株的获得  

Vector Construction of Shoot-specific Promoter Driven Prosystemin and the Acquisition of Transgenic Plants

作　　者：赵雪芹[1,2] 刘维仲[1] 蒋红玲[3] 张海燕[2] 

机构地区：[1]山西师范大学生命科学学院,临汾041000 [2]中国科学院植物研究所分子发育生物学研究中心,北京100093 [3]中国科学院遗传与发育生物学研究所基因组生物学研究中心,北京100101

出　　处：《植物研究》2011年第3期318-322,共5页Bulletin of Botanical Research

基　　金：国家自然科学基金委重大研究计划项目(90817010)

摘　　要：克隆地上部特异表达的启动子——cab2(chlorophyll a/b binding protein 2,cab2)基因的启动子,构建该启动子驱动下的番茄原系统素(Prosystemin;PS)与GFP融合的植物表达载体并获得转基因植株。利用农杆菌介导法转化拟南芥,通过RT-PCR的方法及激光共聚焦显微镜观察启动子驱动PS-GFP的表达及其亚细胞定位。以拟南芥基因组为模板,利用高保真聚合酶获得了cab2启动子的目的片段,并将其与接GFP的番茄原系统素载体(SlPS)融合,激光共聚焦显微镜观察表明,该启动子驱动的基因正常表达和并定位于细胞质中。克隆获得到了cab2基因的启动子,该启动子能够驱动番茄原系统素和GFP的融合蛋白正常表达和定位。The objective of this study was to clone a shoot-specific promoter,it is the promoter of cab2（chlorophyll a/b binding protein 2,cab2） gene.The promoter can drive the expression of prosystemin（PS） and GFP fusion vector,then get the transgenic plants.The plasmids were transformed into Arabidopsis by using Agrobacterium tumefaciens-mediated method.The semi-quantitative RT-PCR method and laser scanning confocal microscope（LSCM） were used to observe the expression and subcellular localization of promoter-driven PS-GFP.Using Arabidopsis genomic DNA as a template,the sequence of promoter cab2 was amplified by high fidelity PCR enzyme,then connected it with the PS and GFP fusion vector（SlPS）.The LSCM observation showed that the gene driven by this promoter was normally expressed and located in the cytoplasm.The cab2 promoter was cloned,which can drive the expression and localization of the PS and GFP fusion protein.

关 键 词：拟南芥 cab2 地上部特异性启动子 系统素 

分 类 号：S641.2[农业科学—蔬菜学]