小麦LEAsm基因及其启动子的克隆与功能研究
发布时间：2018-09-12 20:17
【摘要】：胚胎发育晚期丰富蛋白(LEA)是一个小的高亲水性的蛋白家族,该蛋白家族在逆境胁迫下大量积累,保护植物免受逆境胁迫。LEA蛋白可分为7组,其中重复的11-氨基酸基序是第3组LEA蛋白的特征。为深入分析第3组LEA蛋白在小麦响应逆境胁迫中的作用机制,利用芯片技术从小麦表达谱中筛选出一个渗透胁迫诱导表达的第3组LEA蛋白基因TaLEAsm,然后根据该基因序列设计引物筛选石麦15的BAC文库,获得1个含有该基因的BAC单克隆,以该BAC单克隆质粒为模板,通过BAC延伸测序克隆了TaLEAsm基因及其启动子序列,并对TaLEAsm序列特征、表达模式和启动子功能进行了初步分析。结果表明,TaLEAsm基因序列仅含有1个105bp的内含子,其开放读码框长675bp,编码224个氨基酸。TaLEAsm含有10个11-氨基酸重复序列,属于第3组LEA蛋白。低温、高盐和渗透胁迫均诱导TaLEAsm基因上调表达,但在根和叶中表达模式不同。在TaLEAsm基因起始密码子上游1 500bp序列中,预测含有14个逆境响应顺式元件。在拟南芥中,TaLEAsm基因启动子能够启动GUS基因表达,渗透胁迫诱导GUS基因明显上调表达。以上结果表明,TaLEAsm为小麦脱水响应基因,其启动子为渗透胁迫诱导启动子。
[Abstract]:Late embryonic development rich protein (LEA) is a small high hydrophilic protein family, which accumulates under stress, and can be divided into 7 groups to protect plants from stress. The repeated 11-amino acid motifs are the characteristics of the third group of LEA proteins. In order to analyze the mechanism of the third group of LEA protein in wheat response to stress, A third group of LEA protein gene TaLEAsm, was screened from wheat expression profile by microarray technique. According to the gene sequence, primers were designed to screen the BAC library of Shimai 15. A BAC monoclonal containing the gene was obtained. Using the BAC monoclonal plasmid as template, the TaLEAsm gene and its promoter sequence were cloned by BAC extension sequencing, and the characteristics, expression patterns and promoter functions of TaLEAsm sequence were preliminarily analyzed. The results showed that the Tale Asm gene contained only one intron of 105bp, and its open reading frame was 675 BP, encoding 224 amino acids. Tale Asm contained 10 11 amino acid repeats, belonging to the third group of LEA proteins. Low temperature, high salt and osmotic stress all induced the up-regulation of TaLEAsm gene expression, but the expression patterns were different in root and leaf. In the upstream 1 500bp sequence of the start codon of the TaLEAsm gene, 14 adduct-response elements were predicted. In Arabidopsis thaliana (Arabidopsis thaliana), the promoter of Tale Asm gene could activate the expression of GUS gene, and the expression of GUS gene was up-regulated by osmotic stress. The results indicated that Tale Asm was a dehydration response gene of wheat and its promoter was osmotic stress induced promoter.
【作者单位】： 石家庄市农林科学研究院;
【基金】：国家重点研发计划项目(2016YFD0101802) 国家公益性行业(农业)科研专项(201203012) 河北省科技支撑计划项目(16226320D)
【分类号】：S512.1
[Abstract]:Late embryonic development rich protein (LEA) is a small high hydrophilic protein family, which accumulates under stress, and can be divided into 7 groups to protect plants from stress. The repeated 11-amino acid motifs are the characteristics of the third group of LEA proteins. In order to analyze the mechanism of the third group of LEA protein in wheat response to stress, A third group of LEA protein gene TaLEAsm, was screened from wheat expression profile by microarray technique. According to the gene sequence, primers were designed to screen the BAC library of Shimai 15. A BAC monoclonal containing the gene was obtained. Using the BAC monoclonal plasmid as template, the TaLEAsm gene and its promoter sequence were cloned by BAC extension sequencing, and the characteristics, expression patterns and promoter functions of TaLEAsm sequence were preliminarily analyzed. The results showed that the Tale Asm gene contained only one intron of 105bp, and its open reading frame was 675 BP, encoding 224 amino acids. Tale Asm contained 10 11 amino acid repeats, belonging to the third group of LEA proteins. Low temperature, high salt and osmotic stress all induced the up-regulation of TaLEAsm gene expression, but the expression patterns were different in root and leaf. In the upstream 1 500bp sequence of the start codon of the TaLEAsm gene, 14 adduct-response elements were predicted. In Arabidopsis thaliana (Arabidopsis thaliana), the promoter of Tale Asm gene could activate the expression of GUS gene, and the expression of GUS gene was up-regulated by osmotic stress. The results indicated that Tale Asm was a dehydration response gene of wheat and its promoter was osmotic stress induced promoter.
【作者单位】： 石家庄市农林科学研究院;
【基金】：国家重点研发计划项目(2016YFD0101802) 国家公益性行业(农业)科研专项(201203012) 河北省科技支撑计划项目(16226320D)
【分类号】：S512.1
【参考文献】
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