拟南芥LRR-RLKs亚家族蛋白RLK6的亚细胞定位及RLK6的组织表达
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引用本文:阿依江·哈拜克,杨 敏,韩玉珍.拟南芥LRR-RLKs亚家族蛋白RLK6的亚细胞定位及RLK6的组织表达[J].西北植物学报,2014,34(1):1~6
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作者单位
阿依江·哈拜克1,2,杨 敏1,3,韩玉珍1* (1 中国农业大学 生物学院 植物生理生化国家重点实验室,北京 100193
2 新疆医科大学 药学院,乌鲁木齐 830011
3 河南科技大学 农学院,河南洛阳 471003) 
基金项目:国家自然科学基金(30670192);教育部博士点基金(20120008110018)
中文摘要:以拟南芥(Arabidopsis thaliana)为材料,运用RT-PCR技术扩增得到了富含亮氨酸的类受体蛋白激酶(LRR-RLKs)亚家族基因RLK6,构建了RLK6与绿色荧光蛋白基因(GFP)融合表达载体并转化拟南芥,用激光共聚焦扫描显微镜观察转基因植物细胞表明:RLK6蛋白定位于细胞膜上;将RLK6-GFP在原生质体中进行瞬时表达,进一步证实了RLK6蛋白定位于细胞膜上。构建了RLK6启动子(2 063 bp) 融合GUS报告基因的载体并转化拟南芥,对转基因植株进行组织化学染色分析表明:RLK6在拟南芥的幼苗、根、花、角果等组织中都有表达,花中表达量较高,尤其是在雄蕊中特异高表达,而在茎、莲座叶和干种子中几乎没有表达。RT-PCR分析结果与GUS组织化学染色的结果一致。研究推测,RLK6可能在花器官生长发育或相关生理过程的信号转导中发挥作用。
中文关键词:拟南芥  LRR-RLKs  亚细胞定位  启动子  组织表达
 
Subcellular Localization of RLK6,a Protein of LRR-RLK Subfamily and Tissue Expression Pattern of the RLK6 in Arabidopsis
Abstract:The full length coding sequence without the TAA stop codon of LRR-RLKs gene (RLK6) of Arabidopsis thaliana was cloned by RT-PCR and fused with green fluorescent protein (GFP) gene to construct a plant expression vector.The stable Arabidopsis transformants were observed using confocal laser scanning microscope.The result indicated that RLK6-GFP fusion protein was localized in the plasma membranes.Consistent location was obtained by the experiment of RLK6-GFP transient expression in living Arabidopsis protoplasts.To visualize the temporal and spatial expression patterns RLK6 in plant tissue,a 2 063 bp promoter fragment of RLK6 was fused with the GUS gene to generate transgenic plants.GUS activity was detected in seedlings,roots and flowers,especially in anther,but hardly detected in stems,rosette leaves and dry seeds.Consistent result was obtained by RT-PCR analysis.Overall,these data implied that RLK6 might involve in the signaling pathway of flower organs development or related processes.
keywords:Arabidopsis  LRR-RLKs  subcellular localization  promoter  tissue expression
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