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大豆科学  2015 

大豆GmGST12基因的克隆及表达分析

DOI: 10.11861/j.issn.1000-9841.2015.05.0782, PP. 782-788

Keywords: 大豆,质核互作雄性不育,谷胱甘肽S-转移酶,基因克隆,表达分析

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Abstract:

植物中的活性氧(ROS)作为细胞内或细胞间的信号起重要作用,但其浓度过高可能会导致植物出现雄性不育,而谷胱甘肽S-转移酶类(GSTs)对于解除ROS对细胞的毒害具有重要作用。前期在大豆质核互作雄性不育系NJCMS1A与其保持系NJCMS1B的比较转录组学研究中发现一个差异表达基因GmGST12。本研究通过同源克隆法从NJCMS1A和NJCMS1B花蕾中克隆了GmGST12基因,其编码区序列(CDS)全长均为708bp,且核苷酸序列相同,编码含235个氨基酸的谷胱甘肽S-转移酶;系统发生分析表明,GmGST12与拟南芥AtGSTU9的同源性最高,二者氨基酸序列相似度为52%;组织表达分析表明,GmGST12在NJCMS1A花蕾中的表达水平极显著地高于NJCMS1B花蕾中的表达水平,而在根、茎和叶中的表达水平差异不显著;亚细胞定位结果显示GmGST12定位于细胞核和细胞质中;此外,还构建了植物过表达载体pCAMBIA3301-GmGST12,以用于下一步转基因功能验证研究。以上结果为进一步研究大豆质核互作雄性不育的分子机理提供了基础。

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