摘要
利用Genome Walker方法从巴西橡胶树中克隆了Hb SERK1起始密码子上游1 395 bp的5′调控序列,序列分析表明,该序列A/T含量高达66.2%,符合真核生物启动子序列的特征。Hb SERK1启动子序列中含有多个典型的真核生物启动子基本元件,如:TATA-box,CAAT-box等,同时存在其它应答元件,如:CAT-box、02-site、ERE、ARE、TCA-element、GCN4-motif、ACA-motif等。将Hb SERK1启动子进行5′缺失,并构建了5个植物缺失表达载体。利用真空渗透法和农杆菌介导法将植物缺失表达载体转化烟草,对烟草转化植株进行GUS活性定量分析结果表明,除了SP5外,其它缺失表达载体均可以驱动GUS蛋白的表达,说明在Hb SERK1启动子-1 395^-252 bp区域可以驱动GUS的表达,表明Hb SERK1启动子是具有生物活性的启动子。
The 5＇ regulatory region of HbSERK1 from Hevea brasiliensis was cloned using GenomeWalker strategy. A/T content in this sequence is up to 66.2%, and meet the characteristic of eukaryotic promoter sequence. TATA box, CAAT box CAT-box, 02-site, ERE, ARE, TCA-element, GCN4-motif, ACA-motif were found in this promoter regioru In order to verify HbSERK1 promoter function, Five deletions plant expression vector were constructed and transformed to the tobacco by Agrobacterium-mediated vacuum infiltration method and Agrobacterium-mediated. GUS activity analysis revealed that the region from -1 395-252 bp nt drived expression of the GUS gene except SP5 deletion （-252 bp）. The results of transient expression and stable expression suggest HbSERK1 promoter possess bioactivity.
出处
《热带作物学报》
CSCD
北大核心
2017年第2期295-301,共7页
Chinese Journal of Tropical Crops
基金
海南省自然科学基金(No.314115)
星火计划重点项目(No.2014GA800003)
