构建酚醛树脂为基质的荧光探针应用于细胞中核酸的检测

项目名称： 构建酚醛树脂为基质的荧光探针应用于细胞中核酸的检测

项目编号： No.21505001

项目类型： 青年科学基金项目

立项/批准年度： 2016

项目学科： 数理科学和化学

项目作者： 杨萍

作者单位： 安徽理工大学

项目金额： 21万元

中文摘要： 核酸几乎存在于所有的生物体内，其浓度的异常往往是一些疾病的预兆，因而定量检测生物体内核酸含量十分重要。针对目前出现的以有机染料构建的荧光探针在实际应用中存在难以进入细胞、易被细胞内酶或一些细胞液所降解、检测活细胞内核酸灵敏度低等问题，本项目利用小尺寸的酚醛树脂纳米颗粒作为荧光材料，对目标核酸具有特异性的DNA片段进行标记，并将该片段的另一端与大尺寸金纳米粒子连结，构建短链DNA“捆绑”酚醛树脂到金纳米粒子表面的复合材料。由于金纳米粒子对酚醛树脂的荧光淬灭作用，构建的该类复合材料具有低的荧光发射强度。当目标核酸存在时，由于特殊DNA片段对目标核酸分子的匹配性，标记有酚醛树脂的DNA片段从金纳米颗粒表面脱落，拉远了酚醛树脂与金纳米粒子的距离，酚醛树脂的荧光恢复。与有机染料相比，该类以酚醛树脂为基质构建的荧光探针具有更好的酶切保护作用和更低的生物毒性，能够实现细胞内多个核酸的更灵敏、更有效检测。

中文关键词： 荧光探针；酚醛树脂；活细胞；核酸检测

英文摘要： Nucleic acids exist in almost all organisms. Abnormal concentration of them indicate that disease occurs. Thus, it is important for quantitative detection of nucleic acids in organism. Dye labeled DNA fragment have often been designed as the fluorescence probe for nucleic acid detection in recent years. However, it is difficult for some dye-based probes to pass through plasma membrane. Furthermore, some dye-based probes can be degraded by enzymes in the cells. To resolve these problems, small sized phenol formaldehyde resin (PFR) nanoparticles will be taken as the fluorescence material and specific DNA fragment will be used to labeled PFR nanoparticles. Another end of the DNA fragment will be connected to the large sized gold nanoparticles. By this way, many small sized PFR nanoparticles will be bound onto the surface of Au nanoparticles through the specific DNA fragment. Due to the fluorescent quench from Au nanoparticles to PFR nanoparticles, the fluorescence intensity of PFR nanoparticles becomes lower. However, the DNA fragment will leave Au nanoparticles in the present of the target nucleic acid which can mate with the base sequence, making the distance between PFR nanoparticles and Au nanoparticles much further. Therefore, the fluorescence intensity of PFR nanoparticles will recover. Compared to dyes, the fluorescence probe consisting of PFR nanopartilces has the advantage of better material protection from enzymatic cleavage and lower biotoxicity, which can be applied for the detection of multiple nucleic acids in cells with better sensitivity and effectivity.

英文关键词： fluorescence probe;phenol formaldehyde resin;living cells;detection of nucleic acid