摘要：近年来，荧光标记染料在生.物分析中发挥重要的作用。特别是6’羧基荧光素已经成为检测蛋白质，DNA序列的国际通用的荧光染料。荧光标记法是一种适宜应用在非同位素标记法，因其灵敏度高、且不存在环境污染以及放射性废物处理等问题。已成为目前关注的热点。62116

荧光素及其衍生物具有.高摩尔吸光系数和高荧光量子产率。荧光素具有良好的水溶性、具有羟基、羧基等活性官能团。荧光素具有与被标记化合物形成稳定的共价键的特点。因而被广泛应用在核酸探针研究中。因此，寻找一条适合工业化.生产的合成6-羧基荧光素标记亚磷酰胺化合物的路线至关重要。在已有文献或文件的报道中，荧光素的合成方法是氯化锌催化合成荧光素。这种合成方法虽然简单，但是产物很难提纯，杂质不易除去。因此在本次试验中，我们将甲基磺酸做为溶剂，这样就能很容易除去杂质。但是这种合成方法合成得.到的是5-和6-羧基荧光素异构体的混合物具有相近的物理化学性质不易分离。传统的合成方法是过柱分离，这样只能小批量生产，并且成本较高，不利于工.业化生产。因此本次实验我们将探索出一种能够稳定生产荧光素的化学合成方法，便于运用到实践中去。

毕业论文关键字：荧光标记法；6-羧基荧光素；标记；亚磷酰胺化合物

Abstract In recent years, fluorescent marker dye in the raw. Content analysis plays an important role. Especially the 6 'carboxyl fluorescein has become the detection of protein, DNA sequence of international general fluorescent dyes. Fluorescence labeling method is a suitable for application in the isotope labeling method, because of its high sensitivity, and there is no such problems as environmental pollution and radioactive waste disposal. Has become a hot topic of current. 

With fluorescein. High molar absorption coefficient and high fluorescence when zi chan rate. Fluorescein has good solubility in water, has the active functional groups such as hydroxyl, carboxyl. Fluorescein with marked characteristics of compound to form stable covalent bonds. And therefore is widely used in the study of nucleic acid probe. So, find a suitable for industrialization. Production of the synthesis of 6 - carboxyl fluorescein and phosphoramide compound route is very important. In the existing literature reports or files, fluorescein synthesis method is zinc chloride catalyst in synthesis of fluorescein. The synthetic method is simple, but the product is very difficult to purification, impurity is not easy to remove. Therefore in this experiment, we use methyl sulfonic acid as solvent, so you can easily remove impurities. But the synthetic method to synthesis. To 5 - and 6 - a mixture of carboxyl fluorescein isomers with similar physical and chemical properties of the separation. The traditional synthesis method is a column separation, so only small batch production, and the cost is higher, is not conducive to work. Industry production. Therefore this experiment we will find a way to stable production of fluorescein chemical synthesis method, easy to use to practice. 

Key words: fluorescent tags; 6 - carboxyl fluorescein; Mark; The phosphate amide compounds 

目录

第1章 绪论 5

1.1 引言 5

1.2 荧光素的简介 5

1.3 荧光素的种类 5

1.4 荧光素的主要用途 7

    1.5 荧光素的结构及其特性 7

1.6 荧光素与生物化学 7

1.7 荧光素与生物分析 9