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为了研究杨树AP1同源基因的表达规律,利用PCR技术从毛果杨基因组DNA中克隆出PtrAP1-2基因上游一段2103bp序列。联合使用PLACE和PlantCARE在线软件分析序列,结果表明该序列含有TATA-box和CAAT-box启动子基本元件,另外含有MRE、GT1-motif、AE-box、TCT-motif和Box4多种光响应元件,因此,PtrAP1-2可能与植物开花过程紧密相关。在序列分析的基础上,构建了PtrAP1-2基因启动子驱动GUS报告基因的植物表达载体,命名为PtrAP1-2pro::GUS。利用农杆菌介导的瞬时表达法转化烟草,结果表明PtrAP1-2启动子可以驱动GUS基因在烟草萼片和花瓣中特异表达,在根、茎、叶、雄蕊和心皮等其他组织部位里没有GUS表达活性,这表明PtrAP1-2pro为花器官特异表达启动子。
In order to study the expression pattern of AP1 homologous genes in poplar, a 2103bp sequence upstream of the PtrAP1-2 gene was cloned from Populus tomentosa by PCR. Combining PLACE and PlantCARE online software analysis sequences, the results showed that this sequence contains the basic elements of TATA-box and CAAT-box promoter, MRE, GT1-motif, AE-box, TCT-motif and Box4, Therefore, PtrAP1-2 may be closely related to plant flowering. On the basis of sequence analysis, we constructed a plant expression vector of PtrAP1-2 gene promoter driven GUS reporter gene named PtrAP1-2pro :: GUS. Agrobacterium tumefaciens-mediated transient expression of tobacco transformation results showed that PtrAP1-2 promoter can drive GUS gene specifically expressed in tobacco sepals and petals in the root, stem, leaf, stamens and other tissue parts of the carotid body in the absence of GUS Expression activity, which indicates that PtrAP1-2pro is a flower organ-specific promoter.