遗传 ›› 2009, Vol. 31 ›› Issue (10): 1037-1042.doi: 10.3724/SP.J.1005.2009.01037

• 研究报告 • 上一篇    下一篇

拟南芥CK1A基因功能初步研究

喻达时;赵琼;邓克勤;郭新红

  

  1. 湖南大学生命科学与技术研究院, 化学生物传感与计量学国家重点实验室, 长沙 410082

  • 收稿日期:2009-01-05 修回日期:2009-03-23 出版日期:2009-10-10 发布日期:2009-10-10
  • 通讯作者: 郭新红

Preliminary studies on the function of Arabidopsis CK1A gene

YU Da-Shi;ZHAO Qiong;DENG Ke-Qin;GUO Xin-Hong   

  1. State Key Laboratory of Chemo/Biosensing and Chemometrics, Institute of Life Science and Technology, Hunan University, Changsha 410082, China
  • Received:2009-01-05 Revised:2009-03-23 Online:2009-10-10 Published:2009-10-10
  • Contact: GUO Xin-Hong

摘要:

利用RT-PCR方法从拟南芥中分离了1个CK基因家族成员CK1A, 该基因的ORF全长2 112 bp, 编码一条703个氨基酸残基的多肽。构建了CK1A基因的植物表达载体35S: GFP: CK1A, 采用基因枪法进行的洋葱表皮细胞GFP瞬时表达实验表明, 荧光信号主要分布在细胞核上, 显示CK1A基因的产物可能在细胞核上发挥作用。半定量RT-PCR分析表明: CK1A基因在花中表达量最大, 其次是茎和根, 在叶和叶柄中表达量较弱。蓝光使CK1A基因的表达升高, 12 h时表达量明显增加, 24 h时表达量下降。酵母双杂交结果显示CK1A蛋白在蓝光下能与CRY2蛋白发生相互作用, 暗示CK1A基因可能参与拟南芥的蓝光信号传导途径。

关键词: 酵母双杂交, 基因克隆, 表达分析, CK1A基因, 拟南芥

Abstract:

A casein kinase 1 protein gene, CK1A, was isolated from Arabidopsis seedlings by RT-PCR method. This gene contains an open reading frame of 2 112 bp, which encodes 703 amino acids. The plant expression vector of 35S: GFP: CK1A was constructed by the Gateway System. The 35S: GFP: CK1A fusion protein was localized to the nucleus in onion epidermal cell, indicating that the product of CK1A gene plays a role in the cell nucleus. The semi-quantitative RT-PCR analysis showed that CK1A was highly expressed in flowers, stems and roots, but less in leaves and leafstalks. The yeast two-hybrid analysis demonstrated that CK1A and CRY2 can interact in vivo under blue light, which indicates that CK1A may play an important role in blue light signal induction of Arabidopsis.