【摘要】 通过克隆拟南芥磷酸酶PP2C家族基因At3g51370,构建了绿色荧光蛋白融合表达载体,用基因枪将构建好的载体轰击洋葱表皮细胞进行瞬时表达分析,发现该At3g51370基因表达蛋白定位在细胞核中;用实时定量PCR方法分析At3g51370基因的组织表达特性,发现该基因在花器官中的表达量明显高于其它组织.进一步构建了含At3g51370基因的启动子和GUS报告基因的植物表达载体,经农杆菌介导转化拟南芥,对转基因拟南芥进行GUS组织化学染色,分析该启动子在不同生长时期与不同组织中的转录活性,结果发现,在幼苗期At3g51370基因主要集中在根尖分生组织和顶端分生组织表达,在成年植株中则集中在生殖器官如花和果荚柄等部位表达,在光照和黑暗条件下,At3g51370基因的表达特性没有明显差异.研究表明,At3g51370可能与其它核定位的PP2C磷酸酶一样参与了基因表达的调控,可能在拟南芥早期发育阶段的细胞增值分裂相关信号转导途径中发挥功能,并在花器官的发育过程中行使功能,且不参与光信号转导.更多还原

【Abstract】 A full length cDNA of Protein Phosphatase 2C gene,At3g51370,was fused with green fluorescent protein to construct a plant expression vector.Transient expression analysis indicated that GFP-At3g51370 fusion protein was localized in the nucleus of onion epidermal cells.Real time PCR was used to determine the tissue expression patterns of At3g51370 gene in adult Arabidopsis thaliana.The results showed that there was the highest expression level in the flowers.The promoter At3g51370 was fused to the GUS report gene and transferred into Arabidopsis by Agrobacterium tumefaciens.Gene expression patterns were studied in different growth periods and in different tissues using GUS staining.At3g51370 was expressed in root and apical meristem at the early development stages,while it was mainly expressed in reproductive organs in adult plants.The expression patterns of At3g51370 did not differ between light and dark conditions.These implied that At3g51370 might involve in transcription regulation just as other nuclear localization pp2c genes and functions at the early development stages of Arabidopsis and related to cell reproductive signaling pathway.It also acted during the development of reproductive organs and dids not take part in light signal transduction.更多还原