质子泵在微藻油脂代谢过程中的分子机制研究

项目名称： 质子泵在微藻油脂代谢过程中的分子机制研究

项目编号： No.41406117

项目类型： 青年科学基金项目

立项/批准年度： 2015

项目学科： 海洋科学

项目作者： 张惠莹

作者单位： 清华大学

项目金额： 9万元

中文摘要： 从分子水平揭示油脂合成及调控的机理，改良藻种，控制油脂代谢流向，是当今微藻油脂工程的研究热点。目前有关质子泵在微藻中，特别是在油脂代谢中的作用机制知之甚少。前期的研究发现，碱性pH值可以诱导三角褐指藻和小球藻中的中性脂肪酸含量显著增加，并且质子泵可能在油脂的合成及油质体形成中发挥重要作用。本项目拟在此基础上, 以质子泵和三角褐指藻为主要研究对象：①通过测定碱性pH值诱导下的质子泵活力和油脂含量，分析质子泵在油脂合成中的动态曲线；②根据添加质子泵抑制剂前后转录水平的差异，分析质子泵在pH诱导下参与调控的理化过程；③结合油脂代谢基因组学的相关知识，分析质子泵与油脂合成之间的关联；④通过定量PCR进一步验证质子泵调控油脂代谢的分子机制。作为前瞻性的基础研究，本项目将为深入探究质子泵在微藻油脂代谢中的分子机制，以及科学评价质子泵在油脂代谢中的功能提供理论依据，并为优质产油藻株的构建提供实验基础。

中文关键词： pH值；质子泵；微藻；油脂代谢；分子机制

英文摘要： Currently, the research on microalgae lipid engineering is focused on the molecular mechanism of lipid synthesis, generation of new types of engineering microalgae, and controls on the lipid metabolism flow. However, the role of proton pump in microalgae, especially in the regulation of lipid synthesis, is not clear. The previous study has revealed that microalgae cultures in alkalinous media could induce the lipid accumulation, which was found in Phaeodactylum tricornutum and Chlorella vulgaris. In addition, proton pump seems to function in lipid synthesis and generation of oil bodies. This project use the proton pump and P. tricornutum as the main subjects. The objectives are to (i) through determination of the activity and gene expression of proton pump induced by alkalinous conditions, analyze the dynamic activity and expression of proton pump during the process of lipid accumulation; (ii) by RNA-seq analysis of microalgae transcription level under the inhibition of proton pump activity, analyze the function of proton pump in response to alkalinous conditions; (iii) according to the reference genome of lipid accumulation, analyze the link between proton pump and lipid accumulation; (iv) by means of real time PCR, further verify the molecular mechanism of lipid accumulation regulated by proton pump. As a pers

英文关键词： pH value；Proton pump；microalgae；lipid metabolism；molecular mechanism