拟南芥耐盐相关基因AtSTK的克隆与功能研究

来源 :中国遗传学会第九次全国会员代表大会暨学术研讨会 | 被引量 : 0次 | 上传用户:dfsdfdf
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研究表明,丝氨酸/苏氨酸蛋白激酶是生物酶中的一个大家族,参与环境胁迫信号的传递是其重要功能之一.TaSTK是筛选获得的一个盐诱导后表达增强的小麦基因,通过在拟南芥中的异源功能验证,表明TaSTK基因的过量表达能够显著提高转基因拟南芥的耐盐性.拟南芥丝氨酸/苏氨酸蛋白激酶基因AtSTK是与小麦耐盐相关基因TaSTK高度同源的蛋白激酶基因.本研究对基因AtSTK进行了克隆和功能研究.结果发现,AtSTK基因的过量表达可以明显提高拟南芥对盐、PEG和ABA胁迫的耐受性.生理特性检测表明,AtSTK的过量表达造成拟南芥质膜透性显著降低,脯氨酸含量明显升高,MDA含量下降,这也许是AtSTK过表达造成拟南芥抗逆性提高的生理原因.在高浓度的外源ABA条件下,AtSTK基因的过量表达明显拮抗了ABA对种子萌发的抑制作用。亚细胞定位结果表明,AtSTK普遍分布于细胞质和细胞核。组织表达特异性检测表明,AtSTK基因在拟南芥的生长点具有较强表达。信号通路的RT-PCR检测表明,AtSTK可能主要通过拟南芥的MAPK信号通路传递盐胁迫信号。从拟南芥基因组DNA扩增获得AtSTK基因的全长启动子序列,并根据启动子结构分析结果,对其启动子序列进行了5’端系列缺失。然后在拟南芥中利用GUS报告基因,对这一系列5端序列缺失启动子序列的启动能力进行了检测,同时对其转基因拟南芥200mmo1/L NaCI盐胁迫后的启动能力也进行了检测。最终确定AtSTK基因启动子-99bp~-180bp和-457bp~-779bp区域有增强子作用,位于-115位点的CAAT框可能对基因表达起到了增强作用。启动子片段一旦缺失-29位点的TATA框,会造成其启动下游基因表达的能力丧失。
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