氢气是最好的清洁燃料。绿藻光合产氢是获取氢气的有效途径，但其分子调控机理还不清楚。过去的研究使人们对催化产氢反应酶类及主要过程有所了解，但对起调控作用的因子发掘不足，限制了人们对调控分子机理的认识。因此，本项目采用分子遗传学与定量蛋白质组学相结合策略，从衣藻中发掘产氢调控新基因并研究其功能。首先，从已获得的高产氢衣藻突变体Cr91切入，分离相关基因并进行功能预测。其次，采用已优化的产氢细胞叶绿体及类囊体膜分离纯化等技术，获得不同产氢阶段野生型和突变体上述亚细胞和细胞可溶性组分，并进行蛋白及膜复合体分离。在此基础上，利用目前最灵敏的定量蛋白质组学技术iTRAQ-MS等进行质谱分析，获得与高光合产氢表型相关蛋白数据库及编码基因的完整信息，揭示新基因影响产氢效率的蛋白质分子基础。研究成果不仅有助于加深对绿藻光合产氢分子调控机理的认识，对发掘更多的产氢调控新基因、构建高光效产氢藻株也有重要意义。

Hydrogen gas (H2) represents one of the clean fuel for the future. H2 photoproduction by green algae such as Chlamydomonas reinhardtii is an attractive approach to producing H2 though splitting of water using visible light. However, molecular regulation mechanism of the process is currently not fully understood. Previous studies revealed characteristics of hydrogenases catalyzing the reaction of H2 formation as well as the distinct phases of the process, but protein factors known to be involved in the regulation of the metabolic pathways are still limited. In this project, we make use of a mutant stain (Cr91) of C. reinhardtii, which yields significantly high-H2 -photoproducing phenotype, and characterize its biochemical features using comparative proteomics at cellular and subcellular levels. The specific gene will be cloned and the proteins related to the phenotype will be identified by iTRAQ MS. Experimental data obtained from the research would provide novel sights in terms of the molecular mechanism and valuable gene targets for further bioengineering toward potent strains in large-scale application.