一种精确pH纳米传感器对病毒入侵过程监测的研究

项目名称： 一种精确pH纳米传感器对病毒入侵过程监测的研究

项目编号： No.31300829

项目类型： 青年科学基金项目

立项/批准年度： 2014

项目学科： 生物科学

项目作者： 梁晓声

作者单位： 中南民族大学

项目金额： 20万元

中文摘要： 本申请拟利用mCherry和ferritin轻链融合蛋白组装成纳米笼装结构，在组装过程中使其笼内包装pH敏感的染料分子来构建一种纳米荧光传感器，用于对胞内pH的持续精密监控。传感器中，pH敏感的染料分子做为指示分子，荧光强弱与pH相关。mCherry一方面做为内标荧光，pH数值通过不同荧光比值来确定结果更加精确；另一方面，mcherry对pH不敏感对光漂白敏感，可以使得荧光传感器在使用过程中通过校正消除光漂白的影响，荧光显微镜观察的方法又定性向定量转变。使用纳米笼装结构组装及包装荧光分子后一个纳米颗粒包含多个荧光分子起到一重信号放大作用。另外，铁蛋白轻链内壁富含负电荷对质子有着富集的作用，可以起到另外一重放大作用。通过利用本研究构建的纳米传感器可以精密监控pH依赖的病毒侵入过程。从而揭示病毒入侵的内吞泡酸化速率特征。另外，由于信号的放大，通过本纳米传感器可以用常规荧光显微镜实现单病毒示踪。

中文关键词： 轻链铁蛋白；pH敏感；蛋白纳米笼装结构；；

英文摘要： This application intends to use the mCherry and ferritin light chain fusion protein assembled into nano-cage structure. During the assembly of the cage packaging pH-sensitive dyes were wrapped into the cage. Thus, a nano-fluorescent sensor was built for the continuous precise monitoring of intracellular pH in virus infection prosedure. The pH-sensitive dye molecules as instructions molecules, of which the fluorescence intensity was pH-related. mCherry as the internal standard fluorescence,and as a reference for photobleaching. In this way pH value was determined by the ratio of different fluorescence intensity, which indicated more accurate results.Then, the nanosensor was conneted with virus particle and applied for mornitoring the pH during virus inffection procedure. Nanocages assemblied and packaged of fluorescent molecules,both fluorescent molecules were taken as signal amplification. In addition, inner wall of ferritin light chain was rich in negative charge that have proton enrichment effect,playing another important signal amplification. Through the use of nano-sensors can be constructed in this study, precise monitoring of the pH-dependent viruses. So as to reveal the characteristics of viruses endocytosis bubble acidification rate. In addition, due to signal amplification through the nanosensors sigle

英文关键词： FerritinLE1；pH sensitive；Protein nanocage structure；；