拟南芥PP2A B′调节亚基β亚型抗体的制备及鉴定  

Preparation and Characterization of Anti-Arabidopsis PP2A B′β Antibody

作　　者：周佳琳[1] 张玉兰[2] 母聪聪[2] 袁敏[2] 孙颖[2] 

机构地区：[1]河北师范大学生命科学学院,河北石家庄050024 [2]河北师范大学分子细胞研究所,河北石家庄050024

出　　处：《河北师范大学学报（自然科学版）》2012年第5期512-517,535,共7页Journal of Hebei Normal University：Natural Science

基　　金：国家自然科学基金(30870213)

摘　　要：拟南芥中,蛋白磷酸酶2A(PP2A)B′调节亚基有9个亚型,其中α亚型和β亚型是油菜素内酯(BR)信号通路的正调节子,它们能使转录因子BZR1脱磷酸化.选择β亚型特异的一段多肽P109,利用大肠杆菌表达系统表达并纯化了连有HIS标签和GST标签的融合蛋白HIS-P109和GST-P109.以HIS-P109作为抗原,免疫新西兰兔,获得抗体,然后用GST-P109对抗体进行了亲和纯化.利用此纯化的抗体在不同的拟南芥材料中免疫印迹检测β亚型蛋白的表达,证实制备的抗体能与拟南芥PP2AB′调节亚基β亚型特异性反应,为深入研究PP2A在BR信号转导途径中的功能提供了有力工具.In Arabidopsis there are nine members in the family of PP2A B′ subunit.Among them,α and β isoforms have been identified as the positive regulators in brassinosteroid signaling pathway by dephosphorylating and activating BZR1 transcription factor.Here a specific fragment of 109 amino acids in N-terminal of B′β isoform we selected and expressed as GST-P109 and HIS-P109 fusion protein respectively in E.coli.The purified HIS-P109 was used as antigen to immune rabbit and the purified GST-P109 was used in affinity purification of antibody.Western blot analysis revealed that the resulted anti-β antibody was specific enough in recognizing the known β protein in Arabidopsis.Therefore a powerful tool was obtained for the future study on PP2A function in brassinosteroid signaling pathway.

关 键 词：蛋白磷酸酶2A(PP2A) B′亚基 Β亚型 拟南芥 抗体制备 

分 类 号：Q94[生物学—植物学]