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首页> 美国卫生研究院文献>3 Biotech >Molecular cloning and functional analysis of the promoter of γ-Tocopherol Methyl Transferase (γ-TMT) gene of soybean (Glycine max)
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Molecular cloning and functional analysis of the promoter of γ-Tocopherol Methyl Transferase (γ-TMT) gene of soybean (Glycine max)

机译：大豆γ-生育酚甲基转移酶（γ-TMT）基因启动子的分子克隆和功能分析

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摘要

γ-Tocopherol methyl transferase (γ-TMT) (EC 2.1.1.95) is the key enzyme of the tocopherol biosynthetic pathway that determines the α-tocopherol concentration in plants. The overexpression of γ-TMT has been a successful approach for α-tocopherol enrichment of most plants including soybean. The typical soybean varieties are rich in γ-tocopherol (constitutes nearly 65–70% of its total seed tocopherol pool), while α-tocopherol, the biologically most active form among all tocopherols, constitutes only 10% of the total tocopherol content. The identification of soybean varieties that have seed α-tocopherol as high as > 20% of the total tocopherols has shifted attention towards the breeding based approach for α-tocopherol enrichment of this crop. Previous research on this aspect suggests that polymorphisms in γ-TMT promoter might be associated with the high α-tocopherol concentration of some soybean varieties. To understand the molecular basis of genetic variation for α-tocopherol concentration in Indian varieties of soybean we cloned the 1.4 kb upstream promoter region of γ-TMT from a high α-tocopherol containing soybean variety (Bragg) as well as from a low α-tocopherol containing variety (DS 2706). Cloning of each of these promoters in pORE R2 vector having GUS reporter gene and the subsequent GUS assay revealed a slightly high promoter activity of Bragg γ-TMT as compared to DS 2706 γ-TMT. On promoter sequence analysis, no sequence polymorphisms were observed in the core promoter region of this gene. However, seven single nucleotide polymorphisms (SNPs) were observed outside the core promoter region. Further study based on deletion construct analysis of this promoter will elucidate the significance of these SNPs in influencing the activity of γ-TMT promoter and the α-tocopherol concentration.
机译：γ-生育酚甲基转移酶（γ-TMT）（EC 2.1.1.95）是确定植物中α-生育酚浓度的生育酚生物合成途径的关键酶。 γ-TMT的过表达已成为包括大豆在内的大多数植物中α-生育酚富集的成功方法。典型的大豆品种富含γ-生育酚（占其种子生育酚总量的近65-70％），而α-生育酚是所有生育酚中生物学活性最高的形式，仅占总生育酚含量的10％。种子中α-生育酚含量高达总生育酚> 20％的大豆品种的鉴定，已将注意力转移到基于育种该作物α-生育酚的方法上。先前对此方面的研究表明，γ-TMT启动子的多态性可能与某些大豆品种的高α-生育酚浓度有关。为了了解印度大豆品种中α-生育酚浓度的遗传变异的分子基础，我们从含有高α-生育酚的大豆品种（Bragg）和低α-生育酚中克隆了γ-TMT的1.4kb上游启动子区域。含有生育酚的品种（DS 2706）。与具有DS 2706γ-TMT相比，在具有GUS报告基因的pORE R2载体中克隆了这些启动子中的每一个，并随后进行了GUS分析，发现布拉格γ-TMT的启动子活性略高。在启动子序列分析中，在该基因的核心启动子区域未观察到序列多态性。但是，在核心启动子区域之外观察到七个单核苷酸多态性（SNP）。基于对该启动子的缺失构建体分析的进一步研究将阐明这些SNP在影响γ-TMT启动子活性和α-生育酚浓度方面的重要性。

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