拟南芥CK1A基因功能初步研究
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
网络出版日期: 2009-10-10
Preliminary studies on the function of Arabidopsis CK1A gene
Received date: 2009-01-05
Revised date: 2009-03-23
Online published: 2009-10-10
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。
喻达时,赵琼,邓克勤,郭新红 . 拟南芥CK1A基因功能初步研究[J]. 遗传, 2009 , 31(10) : 1037 -1042 . DOI: 10.3724/SP.J.1005.2009.01037
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.
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