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盐藻小G蛋白基因(DsRab)的克隆及在高盐胁迫下的表达分析

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Abstract:

采用RT-PCR和RACE技术扩增了杜氏盐藻小G蛋白基因cDNA全长序列(GenBankAccessionNo.JN989548),命名为DsRab,对其进行生物信息学分析,并通过实时荧光定量PCR方法检测盐胁迫下该基因的表达情况。结果表明,DsRab基因的cDNA全长为1299bp,开放阅读框(ORF)为612bp,编码203个氨基酸,5'非编码区78bp,3'非编码区609bp;保守性结构域分析可知编码的小G蛋白有4个GTP/GDP保守结构域,1个效应区、1个羧基端的半胱氨酸结构域和5个Rab亚家族共有的结构域;二级结构预测表明该蛋白有32.02%的α-螺旋,23.65%的伸展片段,44.33%的自由卷曲,三维建模成功;比对分析发现DsRab蛋白与多种生物的Ypt/Rab的氨基酸序列具有较高的同源性。荧光定量PCR结果表明,盐藻在高盐(3.0mol/L)胁迫下,DsRab基因表达量显著上调,1h后表达量达到最大值,为正常培养下对照组(0h)的4.9倍,差异极显著(P<0.01)。

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