基于RNA-Seq的裂殖壶菌响应氧因子调控的分子机制研究

项目名称： 基于RNA-Seq的裂殖壶菌响应氧因子调控的分子机制研究

项目编号： No.21306085

项目类型： 青年科学基金项目

立项/批准年度： 2014

项目学科： 化学工业

项目作者： 任路静

作者单位： 南京工业大学

项目金额： 26万元

中文摘要： 二十二碳六烯酸（DHA）是一种重要的ω-3长链多不饱和脂肪酸，具有促进神经细胞发育、预防心血管疾病等功效。近年来，利用微生物产油已成为研究热点。本研究针对产油微生物中不饱和双键形成及碳链延长调控机制不明晰这一问题，以氧刺激可改变裂殖壶菌胞内饱和脂肪酸与不饱和脂肪酸合成效率这一现象为切入点，采用转录组学的研究手段，利用RNA-Seq技术获取在不同氧供应条件及生长产油期裂殖壶菌胞内基因差异表达图谱，结合生物信息学和统计学软件挖掘与不饱和双键形成、碳链延长相关的关键调控基因；构建裂殖壶菌遗传转化体系对关键基因进行过量表达或定点敲除，阐述关键调控因子对裂殖壶菌合成DHA的内在作用机制，揭示氧因子诱导裂殖壶菌胞内饱和与不饱和脂肪酸合成途径偏转的调控机理，发展一条适用于非模式微生物基于转录组样本获取关键调控因子的新路线，对揭示产油微生物不饱和脂肪酸的合成及调控机制具有重要意义。

中文关键词： 裂殖壶菌；二十二碳六烯酸；RNA-seq；氧因子；油脂

英文摘要： Docosahexaenoic acid (DHA) is one of the important ω-3 long-chain polyunsaturated fatty acids, which plays a major role in promoting the development of neurons cells and preventing angiocardiopathy. Recently, microbial oil production has attracted much attention. The problem of the unclear regulation mechanism of double bonds formation and carbon chain enlongation in oleaginous microorganism is concerned in this study. Taking the phenomenon that oxygen could regulate the synthetic efficiency of saturated fatty acids and unsaturated fatty acids as a breakthrough, transcriptomics will be applied to get differential intracellular gene expression profiles of Schizochytrium sp. under different oxygen supply conditions and different fermentation stages. Based on bioinformatics database and statistical software, the key regulatory genes related to double bonds formation and carbon chain enlongation will be achieved. At the same time, genetic transformation system will be developed to knockout or overexpress the specific key genes. The intrinsic mechanism of the key regulatory factor for DHA synthesis by Schizochytrium sp. will also be elaborated. In addition, the regulation mechanism will be revealed to explain how oxygen factor induced the metabolic shift of saturated fatty acid and unsaturated fatty acid pathways in

英文关键词： Schizotrium sp；Docosahexaenoic acid；RNA-seq；oxygen；lipid