基于荧光素和罗丹明的增强型pH荧光探针的设计

发布时间：2018-09-19 07:09

【摘要】：近年来,荧光分子探针因其可视化目标区域和分子的相互作用,而在标记和检测方面被广泛使用,成为了科学探索的重要手段,尤其在医学、环境、生物等领域都展现了更为广泛的应用。鉴于荧光探针在生物医学和环境监测领域的广泛应用,本论文以荧光素、罗丹明为荧光母体,通过对荧光素和罗丹明进行结构修饰,开发一系列可以检测土壤水解酶活性和检测动植物细胞内微环境的pH荧光探针,并测试其相关性质。首先以荧光素为荧光团,在其结构上引入醛基,通过乙酰化设计合成了土壤水解酶活性检测荧光探针A-FDA,其在介质与土壤水解酶相互作用,水解生成荧光素单醛,醛基的引入可以和生物大分子的半胱氨酸结合,减轻了荧光素单醛从细胞中的渗漏。该探针利用水解产物具有强荧光,而A-FDA本身没有荧光这一特性对土壤水解酶的活性进行检测,从而评估土壤中酶活性。由于AFDA的波长与土壤中物质产生干扰。本文在A-FDA的基础上,以半萘荧光素为荧光团,通过乙酰化,设计合成长波的探针SNAFLQ来检测土壤中水解酶的活性。其次以罗丹明、甲硫氨酸、谷氨酸、甘氨酸以及乙醇胺为原料设计了一系列pH荧光探针,分别测试了探针的p H响应、pH可逆性以及生物成像实验,发现甲硫氨酸的探针的pKa值在6.8,偏向于中性,适合于中性条件下的pH检测。最后我们对罗丹明探针进一步进行结构修饰,并测试其对金属离子的响应。
[Abstract]:In recent years, fluorescent molecular probes have been widely used in labeling and detection because of their visualized interaction between target regions and molecules, and have become important means of scientific exploration, especially in medicine and the environment. Biology and other fields have shown wider application. In view of the wide application of fluorescent probes in biomedicine and environmental monitoring, fluorescein and rhodamine were used as the fluorescent matrix in this paper, which were modified by the structure of fluorescein and rhodamine. A series of pH fluorescent probes were developed to detect the activity of soil hydrolase and the microenvironment of plant and animal cells. The fluorescence probe A-FDAwas synthesized by acetylation design using fluorescein as fluorescence group. The fluorescence probe A-FDAwas hydrolyzed to form fluorescein monoaldehyde by interaction with soil hydrolase in the medium. The introduction of aldehydes can bind to cysteine of biomolecules and reduce the leakage of fluorescein monoaldehydes from cells. The probe was used to detect the activity of soil hydrolase by using the characteristic of high fluorescence of hydrolysate and no fluorescence of A-FDA itself, so as to evaluate the activity of soil hydrolase. The wavelength of AFDA interferes with the substance in the soil. On the basis of A-FDA, a long wave probe SNAFLQ was designed to detect the activity of hydrolase in soil by acetylation with naphthalene fluorescein as fluorescence group. Secondly, a series of pH fluorescent probes were designed using Rhodamine, methionine, glutamate, glycine and ethanolamine as raw materials. It was found that the pKa value of methionine probe was 6.8, which was neutral and suitable for pH detection under neutral conditions. Finally, we further modify the structure of Rhodamine probe and test its response to metal ions.
【学位授予单位】：辽宁大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：X833

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