中文摘要：

根据以往试验获得的Cs E1α的c DNA全长序列,利用TAIL-PCR克隆Cs E1α启动子。测序验证与生物信息学分析后发现,该启动子片段长336 bp,含有2个CAAT-box,2个TATA-box,2个GATA-box,1个LTR,1个G-box等顺式作用元件。构建载体转入洋葱内表皮细胞瞬时表达,启动子可启动下游报告基因,使荧光蛋白表达于整个细胞,表明所克隆的启动子具有启动功能。Cs E1α与GFP融合蛋白瞬时表达表明Cs E1α定位于线粒体。本实验为下一步转基因拟南芥稳定表达,进一步研究Cs E1α基因的表达调控,探讨茶树花粉抗寒的分子生物学机理奠定基础。

英文摘要：

In this paper, full-length c DNA is identified for designing the gene-special primers in TAIL-PCR to clone the promoter of Cs E1α. By sequencing and bioinformatic analyzing, we observed two CAAT-box, two TATA-box, two GATA-box, one LTR and one G-box located in the 336 bp promoter region. After constructing and transferring the transient expression vectors into the onion epiderm al cells, subcellular localization of Cs E1α-GFP fusion proteins is verified in mitochondria, while the promoter can activate downstream gene expressing in entire cell. This experiment may provide useful information for the subsequent stable expression in transgenic Arabidopsis and the further investigation on the expression and regulation of Cs E1α gene as well as the investigation on the molecular mechanism of cold resistance in pollen of Camellia sinensis.