拟南芥3个蛋白磷酸酶2C基因编码蛋白的亚细胞定位及组织表达分析

Subcellular Localization and Tissue Expressions of Three Phosphatase 2C Genes in Arabidopsis

利用gateway技术从拟南芥中克隆了3个蛋白磷酸酶2C基因At5G66080、At1G68410和At5G06750,3个基因的ORF全长分别为1 158 bp、1 311 bp和1 182 bp,分别编码一条385、376和393个氨基酸残基的多肽.构建了3个基因的植物表达载体35S:GFP:At5G66080、35S:GFP:At1G68410和35S:GFP:At5G06750,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,显示这3个基因的产物可能在细胞核上发挥作用.利用实时荧光定量PCR研究At5G66080、At1G68410和At5G06750基因在不同组织中的表达特性,结果表明:3个基因在各个器官均有表达,但表达量不同;At5G66080、At1G68410和At5G06750基因在花中表达量最大;At5G66080和At5G06750基因在根、叶和叶柄中的表达量次之,在茎中的表达量最低;At1G68410基因在根中的表达量次之,在茎、叶和叶柄中的表达量较低.

Three protein phosphatase 2C genes including At5G66080,At1G68410 and At5G06750 were cloned from Arabidopsis seedlings by gateway method.The At5G66080,At1G68410 and At5G06750 contain open reading frames（ORF） of 1 158 bp,1 311 bp and 1 182 bp,which encode 385,376,and 393 amino acids,respectively.The plant expression vectors of 35S：GFP：At5G66080,35S：GFP：At1G68410 and 35S：GFP：At5G06750 were constructed by the gateway system.The 35S：GFP：At5G66080,35S：GFP：At1G68410 and 35S：GFP：At5G06750 fusion proteins were localized to the nucleus in onion epidermal cells,indicating that the products of the three genes play a role in the cell nucleus.The quantitative real-time RT-PCR was used to determine the tissue expression patterns of At5G66080,At1G68410 and At5G06750 in different tissues.The results showed that the above three genes were expressed ubiquitously in various organs and tissues,but the expression levels were different.The expressions of At5G66080,At1G68410 and At5G06750 were the highest in the flowers.The expressions of At5G66080 and At5G06750 were lower in root,leaves and leafstalks,and the lowest in the stems.The At1G68410 was expressed lower in root,and the expression levels in the stems,leaves and leafstalk were very low.