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大豆科学  2009 

野生大豆GsADF1基因的克隆与表达分析

DOI: 10.11861/j.issn.1000-9841.2009.05.0784, PP. 784-790

Keywords: 野生大豆,肌动蛋白解聚因子,GsADF,克隆

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Abstract:

ADF蛋白是一种在真核生物中广泛存在的低分子量的肌动蛋白结合蛋白,在细胞分裂、细胞运动、植物顶端生长如花粉管伸长,根毛的形成等重要的生命活动中发挥着重要的作用。以野生大豆(Glycinesoja)的叶片cDNA为模板,通过RT-PCR方法扩增分离获得GsADF1基因。该基因全长为693bp,包含417bp的开放阅读框;其编码的ADF蛋白含139个氨基酸残基。运用生物信息学方法将GsADF1与其他物种ADF蛋白氨基酸序列进行多重比对并构建系统进化树,发现GsADF1与其它生物的ADF蛋白具有很高的同源性。这说明植物ADF基因高度保守。为进一步研究GsADF1在野生大豆不同组织,器官以及不同发育期大豆种子中的表达情况,对它进行了荧光定量分析(RealtimeRT-PCR)。结果表明:GsADF1在野生材料的根、茎、叶、花和种子中都有表达,且以根和花中表达量最高。而对其在不同发育阶段的种子中的分析表明该基因在15DAF、20DAF、30DAF、40DAF和45DAF的种子中都有表达,且以15DAF表达量最低,随后呈上升趋势。其表达量的变化表明GsADF1通过调节肌动蛋白可能参与了种子从胚胎发生到贮藏物质积累的转变。

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