CAJ | 학술논문

根据以往试验获得的CsE1α的cDNA全长序列，利用TAIL-PCR克隆CsE1α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。CsE1α与GFP融合蛋白瞬时表达表明CsE1α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CsE1α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。
근거이왕시험획득적CsE1α적cDNA전장서렬，이용TAIL-PCR극륭CsE1α계동자。측서험증여생물신식학분석후발현，해계동자편단장336 bp，함유2개 CAAT-box，2개 TATA-box，2개 GATA-box，1개 LTR，1개 G-box 등순식작용원건。구건재체전입양총내표피세포순시표체，계동자가계동하유보고기인，사형광단백표체우정개세포，표명소극륭적계동자구유계동공능。CsE1α여GFP융합단백순시표체표명CsE1α정위우선립체。본실험위하일보전기인의남개은정표체，진일보연구CsE1α기인적표체조공，탐토다수화분항한적분자생물학궤리전정기출。
In this paper, full-length cDNA is identified for designing the gene-special primers in TAIL-PCR to clone the promoter of CsE1α. 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 epidermal cells, subcellular localization of CsE1α-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 CsE1α gene as well as the investigation on the molecular mechanism of cold resistance in pollen of Camellia sinensis.