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。
Abstract: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.