生物学杂志 ›› 2019, Vol. 36 ›› Issue (6): 6-.doi: 10.3969/j.issn.2095-1736.2019.06.006

• 研究报告 • 上一篇    下一篇

铁皮石斛氨基酸通透酶基因(DoAAP2)的克隆及表达分析

  

  1. 西南交通大学 生命科学与工程学院, 成都 610031
  • 出版日期:2019-12-18 发布日期:2019-12-12
  • 通讯作者: 王万军,教授,主要从事植物发育生物学研究,E-mail:wanjunwang@home.swjtu.edu.cn
  • 作者简介:刘楚琪,硕士研究生,主要从事植物发育生物学研究,E-mail: 281737941@qq.com
  • 基金资助:
    国家自然科学基金项目(31371232)

Cloning and expression analysis of a amino acid permease gene (DoAAP2) in the Dendrobium officinale Kimura et Migo #br#

  1. School of Life Science and Engineering, Southwest Jiaotong University, Chengdu 610031, China
  • Online:2019-12-18 Published:2019-12-12

摘要:

采用RACE (Rapid amplification of cDNA ends)和巢式 PCR技术克隆出铁皮石斛氨基酸通透酶基因DoAAP2 (Amino acid permease 2)序列。序列分析表明该基因长1772 bp,编码493个氨基酸残基,所编码的蛋白含有完整的Aa trans结构域。系统发育分析显示,DoAAP2与小兰屿蝴蝶兰亲缘关系最近,其次是深圳拟兰,这3种植物同为单子叶兰科植物。实时荧光定量PCR (RT-qPCR) 结果显示,DoAAP2在茎和叶中表达量较高,经过生物信息预测分析亚细胞定位于叶绿体类囊体膜,推测该蛋白可能参与植物的光合作用。在原球茎发育时期中,原分生组织形成(P3)、顶端分生组织形成(P4)、根端分生组织形成(P7)时期表达量较高,表明该基因对铁皮石斛根、茎的发育起重要作用。


关键词: 铁皮石斛, 氨基酸通透酶, RACE, 实时荧光定量

Abstract:

Amino acid permease 2 (DoAAP2) in Dendrobium officinale Kimura et Migo was achieved by RACE (rapid amplification of cDNA ends) and nested PCR technologies. DoAAP2 contained 1772 bp nucleotides with a putative open reading frame encoding 493 amino acid residues with a Aa trans domain. Phylogenetic analysis showed that the DOAAP2 had the closest relation with orchidaceae mono cotyledon plants such as Phalaenopsis equestris and Apostasia shenzhenica. Results of real-time quantitative PCR (RT-qPCR) showed high expression levels of DoAAP2 in leaves and stems, and subcellular localization analysis of bioinformatics prediction showed the product played a major role in the chloroplast thylakoid membrane, suggested that the protein might be involved in photosynthesis. During the development of the protocorm, the expression of primary meristem formation (P3), apical meristem formation (P4) and root meristem formation (P7) was high, indicating that the gene plays an important role in the development of root and stem of Dendrobium officinale Kimura et Migo.


Key words: Dendrobium officinale Kimura et Migo, amino acid permease, RACE, RT-qPCR

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