桂花DAD-1基因克隆与表达分析
发布时间：2018-11-01 20:40
【摘要】：以"柳叶金桂"桂花(Osmanthus fragrans‘Liuye Jingui’)为试材,采用TAIL-PCR技术,克隆了"柳叶金桂"花瓣中DAD-1基因的全长序列,并分析了其在桂花不同组织部位及开花阶段的时空表达模式,以期为研究该基因在桂花花瓣衰老过程中的功能提供参考依据。结果表明:DAD-1基因由560个碱基组成,包含一个351bp的完整开放阅读框和一个3′-Poly A尾巴;DAD-1基因编码的蛋白包含116个氨基酸,其结构预测表明DAD-1蛋白属于疏水性膜整合蛋白,通过3个α?螺旋构成的跨膜区行使功能。半定量PCR结果表明,该基因在"柳叶金桂"的根、茎、叶、花等各组织器官中均有表达,且在花苞、新叶等幼嫩组织器官中相对表达量较高,在老叶等成熟组织器官中相对表达量较低。实时荧光定量PCR结果表明,在"柳叶金桂"花瓣衰老过程中,DAD-1基因在铃梗期开始显著下调表达。
[Abstract]:The full-length sequence of DAD-1 gene in the petals of "Cinnamomum willow" was cloned by means of TAIL-PCR technique. The temporal and spatial expression patterns of the gene in different tissues and flowering stages of Osmanthus fragrans were analyzed in order to provide a reference for studying the function of the gene in the senescence process of Osmanthus osmanthus petals. The results showed that the DAD-1 gene was composed of 560 bases and contained a complete open reading frame of 351bp and a tail of 3'-Poly A. The protein encoded by DAD-1 gene contains 116 amino acids, and its structure prediction indicates that DAD-1 protein belongs to hydrophobic membrane integrin protein. The function of the transmembrane region formed by the helix. The results of semi-quantitative PCR showed that the gene was expressed in root, stem, leaf, flower and so on, and the relative expression was higher in young tissues such as bud and new leaf. The relative expression in mature tissues and organs such as old leaves was low. The results of real-time fluorescence quantitative PCR showed that the expression of DAD-1 gene began to down-regulate significantly during the senescence of the petals of "Cinnamomum willow".
【作者单位】： 湖北科技学院核技术与化学生物学院;孝感市农业科学院蔬菜研究所;华中农业大学园艺植物生物学教育部重点实验室;
【基金】：湖北省教育厅科学技术研究资助项目(Q20162805) 湖北科技学院博士启动基金资助项目(BK1429)
【分类号】：S685.13
本文编号：2305041
[Abstract]:The full-length sequence of DAD-1 gene in the petals of "Cinnamomum willow" was cloned by means of TAIL-PCR technique. The temporal and spatial expression patterns of the gene in different tissues and flowering stages of Osmanthus fragrans were analyzed in order to provide a reference for studying the function of the gene in the senescence process of Osmanthus osmanthus petals. The results showed that the DAD-1 gene was composed of 560 bases and contained a complete open reading frame of 351bp and a tail of 3'-Poly A. The protein encoded by DAD-1 gene contains 116 amino acids, and its structure prediction indicates that DAD-1 protein belongs to hydrophobic membrane integrin protein. The function of the transmembrane region formed by the helix. The results of semi-quantitative PCR showed that the gene was expressed in root, stem, leaf, flower and so on, and the relative expression was higher in young tissues such as bud and new leaf. The relative expression in mature tissues and organs such as old leaves was low. The results of real-time fluorescence quantitative PCR showed that the expression of DAD-1 gene began to down-regulate significantly during the senescence of the petals of "Cinnamomum willow".
【作者单位】： 湖北科技学院核技术与化学生物学院;孝感市农业科学院蔬菜研究所;华中农业大学园艺植物生物学教育部重点实验室;
【基金】：湖北省教育厅科学技术研究资助项目(Q20162805) 湖北科技学院博士启动基金资助项目(BK1429)
【分类号】：S685.13
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