基于熒光素和羅丹明的增強型pH熒光探針的設計
發(fā)布時間：2018-09-19 07:09
【摘要】：近年來,熒光分子探針因其可視化目標區(qū)域和分子的相互作用,而在標記和檢測方面被廣泛使用,成為了科學探索的重要手段,尤其在醫(yī)學、環(huán)境、生物等領(lǐng)域都展現(xiàn)了更為廣泛的應用。鑒于熒光探針在生物醫(yī)學和環(huán)境監(jiān)測領(lǐng)域的廣泛應用,本論文以熒光素、羅丹明為熒光母體,通過對熒光素和羅丹明進行結(jié)構(gòu)修飾,開發(fā)一系列可以檢測土壤水解酶活性和檢測動植物細胞內(nèi)微環(huán)境的pH熒光探針,并測試其相關(guān)性質(zhì)。首先以熒光素為熒光團,在其結(jié)構(gòu)上引入醛基,通過乙�；O計合成了土壤水解酶活性檢測熒光探針A-FDA,其在介質(zhì)與土壤水解酶相互作用,水解生成熒光素單醛,醛基的引入可以和生物大分子的半胱氨酸結(jié)合,減輕了熒光素單醛從細胞中的滲漏。該探針利用水解產(chǎn)物具有強熒光,而A-FDA本身沒有熒光這一特性對土壤水解酶的活性進行檢測,從而評估土壤中酶活性。由于AFDA的波長與土壤中物質(zhì)產(chǎn)生干擾。本文在A-FDA的基礎上,以半萘熒光素為熒光團,通過乙�；�,設計合成長波的探針SNAFLQ來檢測土壤中水解酶的活性。其次以羅丹明、甲硫氨酸、谷氨酸、甘氨酸以及乙醇胺為原料設計了一系列pH熒光探針,分別測試了探針的p H響應、pH可逆性以及生物成像實驗,發(fā)現(xiàn)甲硫氨酸的探針的pKa值在6.8,偏向于中性,適合于中性條件下的pH檢測。最后我們對羅丹明探針進一步進行結(jié)構(gòu)修飾,并測試其對金屬離子的響應。
[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.
【學位授予單位】：遼寧大學
【學位級別】：碩士
【學位授予年份】：2017
【分類號】：X833
本文編號：2249402
[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.
【學位授予單位】：遼寧大學
【學位級別】：碩士
【學位授予年份】：2017
【分類號】：X833
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