Function of the plant gene induced by oxidative stress on the reduciton of lipid hydroperoxide

氧化应激诱导的植物基因对脂质过氧化氢还原的作用

• 批准号: 12660295
• 负责人: SUGIMOTO Manabu
• 金额: $ 1.98万
• 依托单位: OKAYAMA UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2000
• 资助国家: 日本
• 起止时间: 2000 至 2002
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-12660295/
• 关键词: phospholipid hydroperoxide glutathione peroxidase; oxidative stress; salt stress; プロモーター

The gene that includes the promoter region of the phospholipid hydroperoxide glutathione peroxidase (PHGPX)-like protein was cloned from Arabidopsis thaliana. The gene was subcloned into a plasmid, pBI101, resulting the promoter gene was fused with the β-glucuronidase (GUS) gene. The fused gene was transferred to the root of the pea by the particle bombardment system. Transient expression of the GUS was shown in the roots cultured by the MS medium containing 0, 50, and 100mM NaCl. This result suggests that the PHGPX-like gene is not induced by oxidative/salt stress.A cDNA encoding barley PHGPX-like protein was expressed in Escherichia coli cells to produce an extra protein. The PHGPX was not detected in the cell-free extract prepared from the clone cells when tert-butyl hydroperoxide, cumene hydroperoxide, and hydrogen peroxide were used as a substrate. The clone cells were assayed for their ability to grow in the medium containing NaCl. The clone cells were much more tolerant to NaCl stress than the host cells, suggesting that the PHGPX-like protein would be a good candidate component of anti-salt/oxidative enzyme in plants.

从拟南芥中克隆了包含磷脂氢过氧化物谷胱甘肽过氧化物酶（PHGPX）样蛋白启动子区域的基因，该基因被亚克隆到质粒pBI101中，从而使启动子基因与β-葡萄糖醛酸酶（GUS）基因融合。融合基因通过粒子轰击系统​​转移到豌豆的根部，GUS 的瞬时表达如图所示。用含有0、50和100mM NaCl的MS培养基培养的根。该结果表明PHGPX样基因不是由氧化/盐胁迫诱导的。编码大麦PHGPX样蛋白的cDNA在大肠杆菌细胞中表达以产生。当叔丁基过氧化氢、过氧化氢异丙苯、并使用过氧化氢作为底物测定了克隆细胞在含有 NaCl 的培养基中的生长能力。克隆细胞比宿主细胞对 NaCl 胁迫具有更强的耐受性，这表明 PHGPX 样蛋白可能是一种类似的蛋白质。植物抗盐/氧化酶的良好候选成分。

项目成果

Manabu Sugimoto: "Structure and function of a phospholipid hydroperoxide glutathione peroxidase-like protein from barley"Journal of Molecular Catalysis B : Enzymatic. (in press). (2003)

Manabu Sugimoto：“来自大麦的磷脂氢过氧化物谷胱甘肽过氧化物酶样蛋白的结构和功能”分子催化杂志 B：酶学。

Manabu Sugimoto, Kazuyoshi Takeda: "Structure and function of a phospholipid hydroxide glutathione peroxidase-like protein from barley"Journal of Molecular Catalysis B : Enzymatic. (in press). (2003)

Manabu Sugimoto、Kazuyoshi Takeda：“来自大麦的磷脂氢氧化物谷胱甘肽过氧化物酶样蛋白的结构和功能”分子催化杂志 B：酶学。