拟南芥核糖体大亚基L10A的亚细胞定位研究

摘要近期的研究显示，RPL10A蛋白可以穿梭到细胞核内，通过与MYB类似蛋白互作，形成L10-INTERACTING MYB DOMAIN-CONTAINING PROTEIN复合物（LIMYB），从而下调整个翻译系统调节下的基因表达，增强植株对菜豆金色花叶病毒的耐性。这是植物中新发现的抗病毒机制。我们的前期实验已经证实了PVX/AtRPL10A重组病毒接种本氏烟草后，会导致植株矮化及病症加重，并对育性产生重要的影响。RPL10A蛋白质的亚细胞定位将为揭示其受到病毒感染后的抗病机制提供线索。88920

本实验的研究方法是构建目标基因与GFP（绿色荧光蛋白）共表达的载体，然后通过基因枪轰击的方法将质粒导入洋葱表皮细胞中，再利用激光共聚焦显微镜观察绿色荧光融合蛋白在细胞中的位置。由于洋葱表皮细胞较大，容易观察表达部位，可以研究RPL10A的亚细胞定位及移动特性实验。实验结果表明RPL10A主要定位在细胞质和细胞核中，其中细胞质的分布比较多。

Recent studies have shown that RPL10A protein can shuttle into the nucleus and interact with MYB-like proteins to form L10-INTERACTING MYB DOMAIN-CONTAINING PROTEIN complex (LIMYB), which down-regulates gene expression under the regulation of the translation system and enhances plant tolerance of the bean golden mosaic virus。 This is a newly discovered antiviral mechanism in plants。 Our preliminary experiments have confirmed that PVX/AtRPL10A recombinant virus which inoculated to the tobacco can lead to plant dwarf and disease aggravated, and has a significant impact on fertility。 Subcellular localization of RPL10A protein will provide us clues to reveal its resistance to virus infection。

In this study, we constructed the fusion protein expression vector with GFP (green fluorescent protein), then transferred the plasmid into onion epidermal cells by gene gun method, and observed the position of green fluorescence signals in the cells by laser confocal fluorescence microscopy。 As the onion epidermal cells are large, and easy to be observed the expression site, and also can be studied RPL10A subcellular localization and mobility characteristics by the experiment。 The results showed that RPL10A was mainly located in cytoplasm and nucleus, and mainly in the cytoplasm。

毕业论文关键词： RPL10A；亚细胞定位；抗病毒；激光共聚焦荧光显微镜

Key words: RPL10A; Subcellular localization; Antiviral; Laser Confocal Fluorescence Microscopy

目录 3

引言 4

1 材料与方法 5

1。1 实验材料 5

1。2 实验操作 5

1。2。1 基因克隆 5

1。2。2 大肠杆菌转化 6

1。2。3 基因枪的操作 7

1。2。4 GFP荧光的现察 8

2 结果与分析 8

2。1 RPL10A的亚细胞源Q于D优G尔X论V文Y网wwW.yOueRw.com 原文+QQ75201`8766 定位 8

2。1。1 35S:6WT:GFP载体于洋葱表皮中的亚细胞定位 8

2。1。2 35S:6WT:GFP载体于本氏烟草中的亚细胞定位 9

2。2 RPL10A的生物信息学预测 10

2。3实验结论 11

3 讨论拟南芥核糖体大亚基L10A的亚细胞定位研究:http://www.youerw.com/shengwu/lunwen_171643.html