蛋白质亚细胞定位方法的比较
毕业论文关键词：亚细胞定位；免疫荧光；荧光蛋白(GFP)
The Comparison of Protein Subcellular Localization Methods
Abstract:Protein is a macromolecule in organisms, and it is the direct manifestation of biological functions. The proteins can function correctly when they are only transported to their particular subcellular site or secreted to the outside of the cell after they are synthesized. Study the specific location of the protein in cells can improve our understanding of the function of the protein as well as the other proteins which have interactions with it, and it is also conductive to explain the complex life phenomenon. There are many methods of protein subcellular localization，such as immunofluorescence, protein subcellular localization based on the fluorescent, immunohistochemical localization, immune colloidal gold labeling, western blot, etc. This study by immunofluorescence method in the choice of fixative and based on the fluorescent protein subcellular localization in fluorescent protein gene location of the harmony of two methods of experimental conditions to explore, improve the experiment operation process, to determine a more suitable for the method of protein subcellular localization. The experimental results show that the immunofluorescence technique is more suitable for protein subcellular localization.
Key word：Subcellular Localization; Immunofluorescence; Fluorescent Protein(GFP)
目 录
摘要 1
Abstract. 1
引言 2
1.实验仪器、材料及试剂 3
1.1实验仪器 3
1.2实验材料 3
1.3试剂 3
1.4主要试剂的配置 3
2.实验方法与步骤 4
2.1免疫荧光 4
2.2基于荧光蛋白的蛋白质定位 6
3.实验结果 6
3.1免疫荧光固定剂的摸索 6
3.2 荧光蛋白融合位置的摸索 8
3.3 免疫荧光和荧光蛋白方法比较 10
4结论与讨论 11
参考文献 13
致谢 14
蛋白质亚细胞定位方法的比较 引言
蛋白质的功能和它在细胞中的定位有着密切的关系，无论原核还是真核细胞都具有复杂的结构，新生的蛋白质都要被准确的运送到细胞的各个部分才能发挥特定的功能[1]。蛋白质在细胞内的定位，是生物学家们对蛋白质生物学功能进行初步推断的重要依据，也是生物学研究的热门话题，了解蛋白质的亚细胞定位从而对其进行分析并阐明生物学功能，具有重大意义。目前，蛋白质的定位工作已经取得可喜的成果。
各种蛋白质亚细胞定位技术的出现和快速发展，使蛋白质的定位不仅从单个的蛋白质亚细胞定位发展到高通量地进行蛋白质亚细胞定位，而且实现了从静态定位研究到活体动态研究的转变[2]。多种定位方法的综合运用，使得亚细胞定位数据的不断积累完善，为了解蛋白质在细胞内的分布状况、研究蛋白质的功能提供了可靠有力地依据。
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