Plant Biotechnology Research Center, Fudan-SJTU-Nottingham Plant Biotechnology R&D Center, School of Agriculture and Biology, Shanghai Jiao Tong University, 200240 Shanghai, China.
Plant Cell Rep. 2010 Feb;29(2):185-92. doi: 10.1007/s00299-009-0811-2. Epub 2009 Dec 25.
Madagascar periwinkle (Catharanthus roseus) produces many therapeutically valuable terpenoid indole alkaloids (TIAs), such as vinblastine and vincristine derived from the monomers vindoline and catharanthine. Deacetylvindoline-4-O-acetyltransferase (DAT) is a key enzyme for the terminal step of vindoline biosynthesis. In this research, the DAT gene promoter was cloned, sequenced, and analyzed. An approximately 1,773 bp genomic DNA fragment of DAT promoter was obtained. Sequence analysis revealed that DAT promoter contained several potential regulatory elements which were involved in the regulation of gene expression. To investigate its function, the promoter fragments with 5' deletions and gain-of-function deletions were fused to GUS reporter gene, and their expressions were analyzed by transient expression in C. roseus cell suspensions. The regulatory activity of DAT promoter was identified with fluorescence quantitative assays. Three TGACG motifs and one inverted motif (CGTCA) between -808 and -1,086 bp in the DAT promoter were found to be involved in methyljasmonate signal transduction pathway.
马达加斯加长春花（Catharanthus roseus）产生许多具有治疗价值的萜类吲哚生物碱（TIAs），例如长春碱和长春新碱，它们是由单体长春质碱和长春碱衍生而来。去乙酰长春花碱-4-O-乙酰转移酶（DAT）是 vindoline 生物合成的最后一步的关键酶。在这项研究中，克隆、测序并分析了 DAT 基因启动子。获得了约 1773 bp 的 DAT 启动子基因组 DNA 片段。序列分析表明，DAT 启动子包含几个参与基因表达调控的潜在调控元件。为了研究其功能，将启动子片段的 5'缺失和功能获得性缺失与 GUS 报告基因融合，并通过在 C. roseus 细胞悬浮液中的瞬时表达来分析其表达。通过荧光定量测定鉴定了 DAT 启动子的调节活性。在 DAT 启动子的-808 到-1086 bp 之间发现了三个 TGACG 基序和一个反向基序（CGTCA），它们参与茉莉酸甲酯信号转导途径。
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