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草业学报  2013 

拟南芥高亲和性K+载体蛋白基因cDNA克隆及其序列特征分析

DOI: 10.11686/cyxb20130629, PP. 230-238

Keywords: 拟南芥,高亲和性K+载体蛋白,RT-PCR技术,生物信息学分析

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Abstract:

以NaCl胁迫处理的拟南芥幼苗叶片为材料,用RNA提取试剂盒抽提总RNA,通过RT-PCR技术和DNA序列测定分析,证实获得了拟南芥高亲和性K+载体蛋白基因(AtHKT1)的cDNA序列。该cDNA全长1521bp,包括506个氨基酸和1个终止密码子序列,且与原序列(accessionnumberAF237672)同源性为99.34%,但与其他科植物HKT1基因同源性较低,注册该基因到GenBank中,注册号为AY685182。利用生物信息学相关软件分析预测AtHKT1基因蛋白质功能和结构,结果发现,该蛋白分子量为57.45kD,理论等电点为9.33;氨基酸序列中第1~40个氨基酸属信号肽序列;第152~500个氨基酸属TrkH阳离子转运体蛋白保守结构域,并存在蛋白激酶C,酪氨酸蛋白激酶,依赖cAMP/cGMP蛋白激酶磷酸化,糖基化和豆蔻酰化等功能位点;该基因编码的蛋白有10个跨膜结构,N末端、C末端及中部等多个跨膜区具疏水性,符合载体类运输蛋白特点。表明本研究获得了拟南芥AtHKT1基因。

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