减数分裂程序性DNA双链断裂产生相关蛋白质筛选

项目名称： 减数分裂程序性DNA双链断裂产生相关蛋白质筛选

项目编号： No.31301227

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： 张远伟

作者单位： 中国科学技术大学

项目金额： 24万元

中文摘要： 同源染色体的相互识别、配对、联会及重组是减数分裂的核心事件，也是减数分裂同源染色体正确分离的基础，其准确进行则依赖于程序性DNA双链断裂（DSB）的存在。参与减数分裂程序性DSB形成的蛋白质有两类，一类决定DSB能否形成，如SPO11；另一类调控DSB形成的数量及位置，如PRDM9。在低等生物如芽殖和裂殖酵母中，已分别发现14和13种决定减数分裂程序性DSB产生的蛋白，而在哺乳类如小鼠中仅发现4种。那么小鼠还有哪些蛋白质、以及如何决定/调控减数分裂程序性DSB的产生？ 本研究拟用液质联用技术，寻找在第一波生精波中Spo11敲除和野生型小鼠睾丸间差异表达的蛋白质；并利用生物信息学、表达定位、与已知DSB产生关键蛋白间的相互作用分析和条件性基因敲除小鼠等手段，对其进行筛选，以期发现一批新的减数分裂程序性DSB产生功能蛋白，并研究其功能，为相关不育症患者发病机制的探索和诊治提供新思维、新靶点。

中文关键词： 程序性DSB；Spo11；数据库；基因敲除；分子基础

英文摘要： The recognition, pairing, synapsis and recombination between homologous chromosomes, which ensure the correct chromosome segregation in meiotic metaphase I, are the basic features of meiosis. The accurate segregation of homologous chromosomes mainly depends on the programmed DNA double strand breaks (DSBs)generated in leptotene stage of meiosisⅠprophase . Two kinds of proteins are known to be responsible for the initiation of programmed DSB, with one to decide the formation of DSB,such as SPO11, and the other to regulate the sites and numbers of DSBs,such as PRDM9. In lower organisms like budding yeast and fission yeast, 14 and 13 proteins were found respectively,compared with 4 proteins in mice. Thus,it is intriguing to explore which proteins participate in the initiation of programmed DSB except the four known ones and what the functions of these proteins are. In this research, we will search for the proteins differentially expressed between the testes of Spo11 knockout mice and wildtype mice during the first spermatogenic wave by liquid chromatography-mass spectrometry techniques.These proteins will then be sorted by bioinformatics analysis , expression and localization test, protein-protein interactions with proteins that are known to be critical for DSB initiation as well as in vivo examinations using germ

英文关键词： programmed DSB；Spo11；database；gene knockout；molecular basis