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Isolation and Functional Characterization of a Novel Seed-Specific Promoter Region from Peanut

机译:从花生的新种子特异性启动子区的分离和功能表征

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摘要

The importance of using tissue-specific promoters in the genetic transformation of plants has been emphasized increasingly. Here, we report the isolation of a novel seed-specific promoter region from peanut and its validation in Arabidopsis and tobacco seeds. The reported promoter region referred to as groundnut seed promoter (GSP) confers seed-specific expression in heterologous systems, which include putative promoter regions of the peanut (Arachis hypogaea L.) gene 8A4R19G1. This region was isolated, sequenced, and characterized using gel shift assays. Tobacco transgenics obtained using binary vectors carrying uidA reporter gene driven by GSP and/or cauliflower mosaic virus 35S promoters were confirmed through polymerase chain reaction (PCR), RT-PCR, and computational analysis of motifs which revealed the presence of TATA, CAAT boxes, and ATG signals. This seed-specific promoter region successfully targeted the reporter uidA gene to seed tissues in both Arabidopsis and tobacco model systems, where its expression was confirmed by histochemical analysis of the transgenic seeds. This promoter region is routinely being used in the genetic engineering studies in legumes aimed at targeting novel transgenes to the seeds, especially those involved in micronutrient enhancement, fungal resistance, and molecular pharming.
机译:越来越多地强调在植物遗传转化中使用组织特异性启动子的重要性。在这里,我们报告了从花生和拟南芥和烟草种子中的验证分离新种子特异性启动子区。报告的促进子区域称为磨削种子启动子(GSP)赋予异源系统中的种子特异性表达,其包括花生(Arachis Hypogaea L.)基因8a4R19G1的推定启动子区域。使用凝胶移位测定分离,测序和表征该区域。通过聚合酶链式反应(PCR),RT-PCR和揭示塔塔,CAAT盒的存在的基序,CAAT箱,CAAT箱,CAAT箱,CAAT盒的存在的计算分析,确认使用携带由GSP和/或花椰菜马赛克病毒45s启动子的二进制载体获得的二元载体获得的烟草转基因。和ATG信号。这种种子特异性启动子区成功地将报告uida基因靶向拟南芥和烟草模型系统中的种子组织,其中通过转基因种子的组织化学分析证实了其表达。该启动子区域通常用于旨在靶向种子的新型转基因的豆类的基因工程研究,尤其是参与微量营养素增强,真菌抗性和分子药物的豆类。

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