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草业学报  2014 

马铃薯和拟南芥GAPC酶基因的克隆及分析

DOI: 10.11686/cyxb20140129, PP. 239-247

Keywords: 马铃薯,拟南芥,胞质3-磷酸甘油醛脱氢酶,基因克隆,生物信息学分析

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Abstract:

胞质三磷酸甘油醛脱氢酶(GAPC)是糖酵解中的关键酶,近年的研究表明其对盐、低温、高温、氧化、高渗、低磷等多种逆境胁迫均有响应。本实验采用RT-PCR,分别从马铃薯和拟南芥中克隆GAPC的编码区序列(CDS),并作生物信息学比较,同时构建2个基因的植物表达载体。结果表明,StGAPC和AtGAPC2的CDS长度均为1017bp,相似性84%,编码338个氨基酸,相似性为92%。蛋白分子量分别为36.65和36.91kDa,理论等电点为6.34和6.67,均属稳定蛋白;三级结构预测表明两者和水稻(2e5rC)GAPC蛋白结构相似,具有GAPC保守功能域。多种植物GAPC基因的序列进化分析表明同科属植物的同源性较高可归于同一分支,与现有的分类系统相对应。以pBI121为基础载体,构建了由CaMV35S启动GAPC基因的植物表达载体,导入农杆菌EHA105,经PCR检测确定获得阳性克隆。本研究为获得转GAPC基因的材料并进一步研究GAPC在逆境中的功能奠定基础。

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