黄花草木樨MoSOS1基因克隆及表达分析

发布时间：2018-03-01 10:11

  本文关键词： 黄花草木樨 MoSOS基因 同源克隆 序列分析 基因表达　出处：《生物技术通报》2017年09期 　论文类型：期刊论文

【摘要】：植物质膜Na+/H+逆向转运蛋白基因SOS1是植物耐盐性必需的基因之一,在抵御盐胁迫过程中发挥十分重要的作用。以黄花草木樨叶片总RNA为模板,通过RT-PCR结合RACE方法克隆得到黄花草木樨MoSOS1基因全长序列,命名为MoSOS1。序列分析表明该基因全长为3 931 bp,开放阅读框(ORF)为2 874 bp,编码957个氨基酸,分子量为112.8 k D,等电点为5.31。TMHAM软件跨膜区的预测分析表明,黄花草木樨MoSOS1蛋白具有8个跨膜结构区域,N端和C端都位于细胞外。氨基酸序列分析表明,MoSOS1蛋白含有1个Na+/H+Exchanger superfamily和一个c NMP(Cyclic nucleotide-monophosphate)结合位点以及1个CAP_ED(Catabolite gene activator protein-effector domain)superfamily结构域。生物信息预测显示,MoSOS1的编码蛋白为不稳定酸性蛋白,不存在信号肽,二级结构多为α-螺旋和无规则卷曲。荧光实时定量RT-PCR分析表明:随着Na Cl浓度的增加,黄花草木樨地上部和根中MoSOS1基因表达水平呈增加趋势,根中表达量大于地上部,表明MoSOS1基因的表达受盐胁迫诱导和调节。
[Abstract]:Plant plasma membrane Na / H antiporter gene SOS1 is one of the essential genes of plant salt tolerance and plays an important role in resisting salt stress. The total RNA of sweet clover leaves was used as a template. The full-length MoSOS1 gene of Melilotus lutei was cloned by RT-PCR and RACE, and named MoSOS1.The sequence analysis showed that the full-length of the gene was 3 931 BP, and the open reading frame (ORF) was 2 874 BP, encoding 957 amino acids. The predicted molecular weight is 112.8 KD and the isoelectric point is 5.31.TMHAM. The N-terminal and C-terminal of the MoSOS1 protein are located outside the cell. Amino acid sequence analysis shows that the moosos1 protein contains a Na / H Exchanger superfamily and a c NMP(Cyclic nucleotide-monophosphate) binding site and a CAP_ED(Catabolite gene activator protein-effector domain)superfamily node. Domain. Bioinformatics prediction shows that the encoded protein of MoSOS1 is unstable acidic protein. There was no signal peptide, and the secondary structure was mostly 伪 -helix and irregular curl. Fluorescence real-time quantitative RT-PCR analysis showed that with the increase of NaCl concentration, the level of MoSOS1 gene expression in the upper part and root of Xidi showed an increasing trend. The expression of MoSOS1 gene in root was higher than that in shoot, which indicated that the expression of MoSOS1 gene was induced and regulated by salt stress.
【作者单位】： 北京市农林科学院北京草业与环境研究发展中心;
【基金】：国家国际科技合作专项(2015DFR30570) 北京市农林科学院科技创新能力建设专项(KJCX20170110)
【分类号】：Q943.2
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本文编号：1551473