拟南芥细胞中存在中间纤维的研究  

The Existence of Intermediate Filament in Arabidopsis thaliana Cell

作　　者：赵大中[1] 陈丹英 陈敏[1] 刘春香 杨澄[1] 翟中和[1] 

机构地区：[1]北京大学生命科学学院,北京100871 [2]中国科学院植物研究所,北京100093

出　　处：《Acta Botanica Sinica》1999年第8期795-799,共5页Acta Botanica Sinica（植物学报：英文版）

摘　　要：利用整装电镜制样与选择性抽提技术，在拟南芥（Ａｒａｂｉｄｏｐｓｉｓｔｈａｌｉａｎａ （Ｌ．） Ｈｅｙｎｈ） 愈伤组织细胞质中观察到直径１０ ｎｍ 左右的纤维网络结构。免疫印迹分析表明纤维的主要成分是６ 种多肽，它们分别与动物角蛋白单克隆抗体ＡＥ１ 、ＡＥ３ 有免疫交叉反应。利用间接免疫荧光技术，与ＡＥ１ 和ＡＥ３ 反应的抗原呈弥散状定位于整个细胞质中，而且１０ ｎｍ 纤维可以在体外重新组装。以上结果表明，在拟南芥细胞质中存在类角蛋白的中间纤维。以动物中间纤维基因的保守序列为引物，采用ＲＴ＿ＰＣＲ技术，进一步从这一模式植物中克隆到一个ｃＤＮＡ片段。The network structure of cytoplasmic filaments of 10 nm in diameter was detected from callus cells of Arabidopsis thaliana (L.) Heynh by selective extraction combined with whole mount electron microscopy. Western blot analysis showed that the major filament components were 6 polypeptides, which reacted with keratin monoclonal antibody of AE1 or AE3 respectively. By indirect immunofluorescence technique, the AE1 and AE3_reactive antigens were localized throughout the cytoplasm in a diffused pattern. The 10 nm_plant filaments could be reassembled in vitro. These results demonstrated that keratin_like intermediate filaments exist in the cytoplasm of A. thaliana . Using conservative sequence of animal IF genes as primer, a cDNA fragment was further cloned from this model material by RT_PCR, which might shed more light on molecular characterization of IF existence in higher plant.

关 键 词：拟南芥 中间纤维 选择性抽提 免疫印迹反应 

分 类 号：Q942.4[生物学—植物学]