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- 2016
阳春砂HMGR基因cDNA克隆及生物信息学分析
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Abstract:
摘要 目的克隆阳春砂3-羟基-3-甲基戊二酰辅酶A还原酶(HMGR)基因的编码区cDNA.方法:根据阳春砂叶片转录组得到的HMGR基因序列自主设计引物.用改良的CTAB法提取阳春砂叶片总RNA,反转录得到cDNA,以cDNA为模板通过PCR反应扩增阳春砂HMGR基因并克隆到pMD-18T载体,将获得的阳性克隆进行测序分析.运用生物信息学方法分析得到的cDNA序列.结果:成功扩增出全长1 716 bp的cDNA片段,将其成功克隆进入pMD-18T载体,测序后将序列提交至GenBank得到其编号KU572444.生物信息分析结果显示,该基因片段编码1个由571个氨基酸残基组成HMGR蛋白.HMGR二级结构主要由43.95%的α-螺旋和47.29%的β-转角组成,同时还含有8.76%的β-折叠.与其他植物该蛋白质序列同源性及进化树分析结果表明,阳春砂HMGR与小米(Setaria italic)的同源性最高,且阳春砂与单子叶植物聚为一类.结论:从阳春砂叶片中成功克隆HMGR基因并进行了生物信息学分析.
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