毛白杨PtSEP3-1基因启动子的克隆分析及其表达载体构建

Cloning and Construction of Expression Vectors of PtSEP3-1 Promoter from Populus tomentosa

SEP(SEPALLATA)类基因属于花器官发育ABCDE模型中的E类基因,拟南芥中的研究表明该类基因可能具有控制花器官形态发育以及激活其它类型基因的功能,是一类花发育过程中的关键基因。因此,研究杨树SEP类基因启动子表达特性对于杨树的开花调控研究具有重要意义。本文根据毛白杨SEP3基因和毛果杨基因组序列设计引物,通过PCR获得了PtSEP3-1基因上游2000bp的序列。序列分析结果表明该序列具有启动子的基本元件TATA-box和CAAT-box,还包含大量光响应元件ACE、Box I和Box4等,此外还有脱落酸响应元件ABRE,赤霉素响应元件GARE-motif以及胁迫响应元件HSE、TC-richrepeats等。进一步构建了一个以PtSEP3-1启动子驱动GUS基因的植物表达载体pPtSEP3-1protest,为该启动子的功能鉴定奠定了基础。

SEP （SEPA LLA TA）-class genes confer E-function in ABCDE model that depicts its roles in floral organ development. Previous studies on Arabidopsis have shown that, genes of this class, key component during floral development, might feature the function of determing floral organogenesis and activating other genes. Therefore analyse expression characteristics of the poplar SEP-class genes promoter are significant for its regulation of flowering. Here, a 2 000 bp long sequence that locates at upstream region of PtSEP3-1 was isolated by using PCR with primers that designed from genomic sequences ofP. trichocarpa and SEP3 of P. tomentosa. Sequence analysis revealed this sequence contains basic promoter elements, TATA-box and CAAT-box. Light response cis-acting elements, ACE, Box I, Box 4, ABA response element ABRE, GA response elements GARE-motifand stress-induced cis-acting elements including HSE, TC-rich repeats were also represented. Moreover, PtSEP3-1 protest vector carrying PtSEP3-1 promoter：GUS was constructed, laying a foundation for functional identification of the PtSEP3-1 promoter.