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利用PCR技术从毛白杨基因组DNA中扩增获得花器官发育相关的SEPALLATA2类似基因PtSEP2 5’侧翼约2.3kb的一段序列,经PlantCARE序列分析表明,该序列中含有启动子特征的保守序列及多种光应答元件,初步推测其为PtSEP2基因启动子。进一步以GUS为报告基因,构建了pPtSEP2 promoter∷GUS的植物表达载体,命名为PtSEP2p∷GUS。以烟草根、茎、叶、花芽为受体,通过农杆菌介导转化植物表达载体PtSEP2p∷GUS进行瞬时表达研究,结果表明该启动子能够驱动GUS报告基因在烟草花药中表达,而其表达活性弱于组成型表达的花椰菜花叶病毒(CaMV)35S启动子。该启动子的获得对于杨树及其他开花调控基因工程研究具有重要的应用价值。
The PCR-based technique was used to amplify a sequence of about 2.3 kb flanking 5 ’of the SEPALLATA2-like gene related to floral organ development from Populus tomentosa genomic DNA. PlantCARE sequence analysis showed that this sequence contains conserved sequences of promoters and multiple Light response element, presumed to be PtSEP2 gene promoter. Further, using GUS as a reporter gene, a plant expression vector of pPtSEP2 promoter :: GUS was constructed and named as PtSEP2p :: GUS. The tobacco roots, stems, leaves and flower buds were used as recipients and transiently expressed in Agrobacterium tumefaciens-mediated plant expression vector PtSEP2p :: GUS. The results showed that the promoter could drive GUS reporter gene expression in tobacco anthers, and its expression activity Weaker than the constitutively expressed cauliflower mosaic virus (CaMV) 35S promoter. The promoter has important application value for poplar and other flowering control gene engineering.