基于微卫星序列[TTAGGG]n的C2H2型锌指结构域的设计

项目名称： 基于微卫星序列[TTAGGG]n的C2H2型锌指结构域的设计

项目编号： No.21301050

项目类型： 青年科学基金项目

立项/批准年度： 2014

项目学科： 数理科学和化学

项目作者： 赵东欣

作者单位： 河南工业大学

项目金额： 25万元

中文摘要： 人染色体端粒末端的[TTAGGG]n重复DNA序列是基因组中重要的微卫星家族之一，影响着染色体的结构、基因的转录与翻译等过程，与多种癌症相关。因此，获得能与[TTAGGG]n特异性结合的分子是治疗肿瘤的一种有效途径。锌指蛋白是人体内一类重要的功能蛋白，通过与 DNA结合而参与调控众多生理过程，其中的C2H2型锌指结构域结构上相对保守，DNA识别模式也相对简单，为设计识别特定DNA序列的功能分子提供了有用的元件。而基于分子片段的筛选法，为锌指的设计提供了新思路。本项目拟以[TTAGGG]n为靶点，以C2H2锌指对碱基的基本识别为着眼点，设计与靶点结合的锌指；然后用筛选法模拟其与靶点的相互作用，再合成出相互作用较强的锌指，并对其进行细致的结构改造，同时通过NMR、MS、CD等方法研究锌指及与靶点的相互作用，为C2H2锌指蛋白的研究、开发和应用提供更详细的实验数据，为基因治疗提供更广阔的前景。

中文关键词： 固相合成；锌指肽；圆二色谱；荧光光谱；模拟

英文摘要： The repeat DNA array [TTAGGG]n which lies in the terminator of human chromosome telomere is one important of microsatellite families in human genome, and directly affects the structure of chromosome, some processes like gene transcription and translation, and are correlated with various cancers. Therefore, the molecular which can specially bind with [TTAGGG]n is a kind of potential medicament for tumors. Zinc finger protein is a type of important function protein inside the human body, and can adjust many physiological progresses through binding with DNA. The C2H2 zinc finger domain has a conservative structure and a simple recognition mode to DNA, which provides a useful mode and component for designing function molecular to identify the particular DNA sequence. And the method of fragment-based screening (FBS) is a new line of thinking for design of zinc finger peptide. This project plans to use the [TTAGGG]n as target and base on the basic recognition mode of C2H2 zinc finger domain with DNA to design and synthesize the C2H2 zinc finger domains which can interact with [TTAGGG]n, then use FBS to simulate the interaction between the designed zinc finger and target, and synthesize the zinc finger domains which have stronger interaction with [TTAGGG]n and reform the structure of zinc finger domains. In this proces

英文关键词： Solid phase synthesis；Zinc finger peptide；Circular dichroism spectrum；Fluorescence spectrum；Simulation