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水稻泛素缀合酶基因OsUbc13的分子特征和蛋白互作研究
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水稻泛素缀合酶基因OsUbc13的分子特征和蛋白互作研究
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Journal Article
水稻泛素缀合酶基因OsUbc13的分子特征和蛋白互作研究
2014
Overview
研究目的：通过研究水稻泛素缀合酶基因OsUbc13的序列特征、表达模式、亚细胞定位模式及其互作分子，为深入研究该基因的生物学功能和分子作用机理奠定基础。创新要点：首次对植物Ubc13进行了亚细胞定位研究及蛋白互作研究。研究方法：通过序列比对及聚类分析进行OsUbc13的序列特征研究；通过实时荧光定量聚合酶链式反应（PCR）进行OsUbc13的表达模式分析；通过聚乙二醇（PEG）介导转化烟草BY-2原生质体进行OsUbc13亚细胞定位研究（见图4）；通过酵母双杂交进行OsUbc13的蛋白质互作分析（见图5和表1）。重要结论：OsUbc13编码具有153个氨基酸的蛋白质，其推断的氨基酸序列与其它同源序列具有很高的相似性；该基因在水稻各组织中均有表达，其中内稃、雌蕊、雄蕊和叶片中的表达量较高，而根、茎和外稃中的表达量较低；低温、甲基磺酸甲酯（MMS）和过氧化氢（H2O2）胁迫处理使胚性愈伤中OsUbc13的表达量显著上调，甘露醇、脱落酸（ABA）和氯化钠（NaCl）胁迫则使愈伤组织中该基因的表达量降低；OsUbc13与绿色荧光蛋白（GFP）的融合蛋白表达于质膜和核膜处；酵母双杂交结果表明约有20个蛋白可能与OsUbc13存在相互作用，其中OsVDAC（与细胞凋亡有关）、OsMADS1（与花器官发育有关）、OsB22EL8（与活性氧清除及DNA 保护有关）和OsCROC-1（为Lys63聚合泛素链形成及运行无误性DNA 损伤耐受机制所必需）四个蛋白经验证确与OsUbc13互作。
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