极地海洋嗜冷菌适冷应答的转录调控网络研究

项目名称： 极地海洋嗜冷菌适冷应答的转录调控网络研究

项目编号： No.41476166

项目类型： 面上项目

立项/批准年度： 2015

项目学科： 海洋科学

项目作者： 刘贵明

作者单位： 中国科学院北京基因组研究所

项目金额： 88万元

中文摘要： 微生物是极地生态系统的重要组成部分,由于极地独特的酷寒、强辐射和高盐度等特点，极地微生物所具有的嗜冷、嗜盐、耐辐射和耐压特性，在科学研究、应用开发等方面都具有重要价值，已成为微生物学研究的热点之一。在前期RNA-seq 研究中，我们发现转录因子在适冷应答中具有显著差异表达。本项目拟以已完成基因组测序和注释的嗜冷菌G.psychrophila 170T为研究对象，从转录组水平研究不同温度梯度应激条件下的转录因子(TF)、mRNA和sRNA 应答响应；基于含Myc抗体标签和荧光蛋白的λ噬菌体Red系统，高通量利用抗体标签和荧光蛋白进行chip和流式分选，分析温度胁迫应答相关TF 与sRNA和mRNA 作用关系，绘制TF在适冷应答中的调控网络。利用基因敲除和过表达技术，鉴定转录因子和靶基因功能,以期建立嗜冷菌的转录因子冷适应调控网络模型，为嗜冷调控机制研究奠定理论基础。

中文关键词： 嗜冷菌；非编码RNA；转录因子；调控网络；chip-seq

英文摘要： Microbiology is an important part of polar ecosystems, because of its frosty, intense radiation and high salinity, polar microbiology have the characteristic of psychrophilic, halophile, radiation resistance and pressurization. They have important value in scientific research and application development, which make them research hotspots in the field of microbiological research. During the preliminary study of RNA-seq at different temperature, we found that some transcript factors are highly different expression. With the recent finished G.psychrophila 170T genome as model bacteria, this project aims to investigate the cold response of transcrip factor(TF),mRNA and sRNA using transcriptome at different temperature. Based on the λ phage Red recombination system with Myc tag antibody and fluorescent protein,we can high-throughput perform chip and flow sorting to study the interaction relationship between TF,mRNA and sRNA,then construct the interaction network of TF with them. Gene knockout of Red recombination system and overexpression are used to study the function of TF and its target genes so as to provide a theoretical foundation for research of mechanism of bacterial cold-adaptation.

英文关键词： psychrophilic bacterium;sRNA;transcript factor;regulatory network;chip-seq