毛白杨PtSEP2基因启动子克隆和瞬时表达特性分析  

Cloning and Transient Expression Analysis of PtSEP2 Promoter from Populus tomentosa

作　　者：崔东清[1] 叶梅霞[1] 刘军梅[1] 李昊[1] 张志毅[1] 安新民[1] 

机构地区：[1]北京林业大学林木育种国家工程实验室林木花卉遗传育种教育部重点实验室国家林业局树木花卉育种与生物工程重点开放实验室北京100083

出　　处：《中国生物工程杂志》2011年第5期42-47,共6页China Biotechnology

基　　金：国家自然科学基金(30571511);国家林业公益性项目(201004009);教育部重点项目(108017)资助项目

摘　　要：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。A 5′ flanking sequence of PtSEP2 which is a SEPALLATA2-like gene involved in floral organ development was amplified by PCR from the genomic DNA of Populus tomentosa.The length of fragment is approximately 2.3kb.The results derived from PlantCARE analysis showed that the sequence contains conserved cis-acting elements of promoter,coupled with a variety of light-responsive elements.Therefore,it was speculated preliminarily to be the promoter of PtSEP2 gene.To investigate function of this promoter,it was fused to GUS reporter gene,generating a plant expression vector pPtSEP2 promoter∷GUS,named PtSEP2p∷GUS.With roots,stems,leaves and flower buds of Nicotiana tobaccum for the receptor,the results of transient expression that Agrobacterium-mediated that showed the PtSEP2 promoter was competent to drive specially GUS reporter gene expression in anther,yet its’ activity was weaker than that of cauliflower mosaic virus（CaMV） 35S promoter,which constitutively expressed in planta.Consequently,provides a possible genetic modification tool for flowering regulation in poplar and other plants.

关 键 词：毛白杨 SEPALLATA2启动子 GUS活性 瞬时表达 

分 类 号：Q786[生物学—分子生物学]