拟南芥磷酸酶基因亚细胞定位与组织表达.pdf

Aeta Bot．Boreal．-Occident．Sin．文章编号：1000-4025(2009)02—0234—06 拟南芥磷酸酶基因亚细胞定位与组织表达邓克勤，郭新红，汪启明，刘选明。(湖南大学生命科学‘J技术研究院生物能源勺材料研究中心，长沙410082) 摘要：通过克隆拟南芥磷酸酶PP2C家族基因At3951370。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At3951370基因表达蛋白定f讧在细胞核中；用实时定量PCR方法分析At3951370基冈的组织表达特性。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At3951370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(；us组织化学染色，分析该启动子存不同牛长时期与不同组织中的转录活性。结果发现．在幼苗期At3951370基冈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化表达，在光照和黑暗条件下，At3951370基闪的表达特性没有明显差异。研究表明，At3951370Ⅱr能与其它核定位的PP2C 磷酸酶一样参与了基闪表达的调控，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。关键词：拟南芥；磷酸酶；亚细胞定化；启动子；组织表达中图分类号：Q789 文献标识码：A Subceilular Localization and Tissue Expression Pattern of Arabidopsis Phosphatase Gene DENG Ke—qin，GU()Xin—hong，WANG Qi—ruing，LIU Xuan—ming。(Institute of Lire Science and Technology．Bioenergy and Biomaterial Research Center．Hunan University．Changsha 410082，China) Abstract：A full length eDNA of Protein Phosphatase 2C gene，At3951370，was fused with green fluorescent protein to construct aplant expression vector．Transient expression analysis indicated that GFP—At395 1370 fusion protein was localized inthe nucleus of onion epidermal cells．Real time PCR was used todetermine the tissue expression patterns of At3951370 gene inadult Arabidopsis thaliana．The results showed that there was the highest expression level in the flowers．7Fhe promoter At3951370 was fused to the GUS re—port gene and transferred into Arabidopsis by Agrobacterium tume{aHens．Gene expression patterns were studied in different growth periods and in different tissues using GUS staining．At395 1370 was expressed in root and apical meristem atthe early development stages，while itwas mainly expressed in reproductive or—gans inadult plants．The expression patterns of At3951370 did not differ between light and dark condi—tions．These implied that At3951370 might involve in transcription regulation just as other nuclear localiza—tion pp2c genes and functions atthe early development stages of Ar