极端嗜盐古菌盐胁迫相关非编码RNA的鉴定和调控机制研究

项目名称： 极端嗜盐古菌盐胁迫相关非编码RNA的鉴定和调控机制研究

项目编号： No.31301071

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： 王磊

作者单位： 中国科学院微生物研究所

项目金额： 24万元

中文摘要： 非编码RNA在细菌、古菌及真核生物中均发挥着至关重要的作用，他们参与了细胞内多种基础生物过程，包括转录调控、RNA加工、翻译调控等。科学家们已经在真核生物和细菌中鉴定了大量非编码RNA，但是古菌中鉴定新非编码RNA方面的研究却很少。本项目以自主完成全基因组序列测定的极端嗜盐古菌- - 地中海富饶盐杆菌为研究对象，通过链特异性RNA-seq和小RNA测序来鉴定其基因组范围内的非编码RNA；通过不同盐浓度样本的比较来鉴定与盐胁迫相关的非编码RNA，并通过Northern Blot加以验证；构建非编码RNA基因缺陷型菌株，利用Microarray分析探索非编码RNA参与基因表达调控的机制。本项目将为嗜盐古菌非编码RNA的研究提供基础数据，并为后续非编码RNA对基因调控机制的研究提供新的思路和见解。

中文关键词： 极端嗜盐古菌；非编码RNA；盐胁迫；基因调控；转录组测序

英文摘要： Non-coding RNAs (ncRNAs) play vital roles in all three domains of life, i.e. Eukarya, Bacteria and Archaea. They participate in many different basic biological processes in the cell including transcriptional regulation, RNA processing, translational regulation. A great diversity of ncRNAs with different roles have been intensely studied in eukaryotic and bacterial model organisms, but few attempts have been performed so far to identify novel ncRNAs in the domain of Archaea. Using strand-specific RNA sequencing and small RNA sequencing methods, this project will identify the genome-wide ncRNA in the haloarchaeon Haloferax mediterranei whose whole genome has been sequenced by our lab. The ncRNAs in response to salt stress will also be identified and confirmed by Northern analysis. Furthermore, this project will construct ncRNA gene deletion mutants and research the regulation mechanism of ncRNAs with microarray analyses. Taken together, this project will provide basic information for the research of ncRNAs in the haloarchaea and provide novel methods and insights in gene regulation mechanism of ncRNAs.

英文关键词： Haloarchaea；non-coding RNA；salt stress；gene regulation；RNA-seq