利用植物悬浮培养细胞进行蛋白质快速定位分析的方法  

Analysis of Protein Subcellular Localization by Using Plant Suspension Cultured Cells

作　　者：赵蒙晴 高野哲夫[2] 柳参奎[1] 

机构地区：[1]东北油田盐碱植被恢复与重建教育部重点实验室,东北林业大学盐碱地生物资源环境研究中心,哈尔滨150040 [2]东京大学亚洲生物资源环境研究中心,东京1880002

出　　处：《基因组学与应用生物学》2012年第4期389-393,共5页Genomics and Applied Biology

基　　金：国家林业局948项目(2008-4-29);黑龙江省杰出青年科学基金(JC200609)共同资助

摘　　要：稳定遗传表达分析是一种植物中常用的整体解析基因的方式。有多种转化方式可供选择,也可根据所需要的获得的转基因植物材料选择受体材料。但是由于稳定遗传转化周期较长且大部分材料不适合于进行荧光观察,所以在一些基因的研究中逐渐被瞬时表达分析系统。虽然瞬时表达分析用时短,但是转化效率受到多方面的限制,转化材料无法保存。目前由于植物悬浮培养细胞材料均一,增殖迅速并且可以满足大批量研究需求逐渐成为植物研究中的热点材料。以此同时,在亚细胞定位方面,悬浮培养细胞还是良好的应用材料。采用农杆菌介导法进行植物悬浮培养细胞的转化中方法较为成熟,但是获得纯净的转基因细胞系的转化周期较长。在本研究中针对上述问题我们建立了一种转化时间短,转化效率高的植物悬浮培养细胞稳定遗传转化体系。同时将这个体系应用到基因的亚细胞定位当中进行蛋白质快速定位分析。Stable genetic expression assay is a common way to analyze genes in plants as a whole and is recognized as a standard to confirm the characteristics of gene function. There are various methods of transformation, and the receptors can also be chosen according to the transgenic plants. However, the stable genetic transformation takes too much time and most of the materials are not suitable for fluorescence observation, so transient expression analysis system is gradually used to study the genes. Although the transient expression analysis takes a shorter time, the transformation efficiency is restricted in many respects, and the transformation material can not be conserved. Because plant suspension cultured cells are uniform and proliferate rapidly, they meet the research needs of large quantities and gradually become hot materials in plant research. Meanwhile, in subcellular localization, suspension culture cells are good application materials. The plant suspension cultured cell transformation mediated by Agrobacterium is more mature, but it takes a long time to obtain a pure transgenic cell line. In this study, a stable genetic transformation system of the plant suspension culture cells is established, and ithas short transformation time and high transformation efficiency. This system is also applied into the subcellular localization of the gene to do the fast protein localization analysis.

关 键 词：悬浮培养细胞 稳定遗传转化 高效 快速 GFP 亚细胞定位 

分 类 号：Q943[生物学—植物学]