| Summary: | 碩士 === 國立中央大學 === 生命科學研究所 === 98 === Biodiesel is an alternative energy source and a substitute for petrochemical diesel fuels. Transferification of triacylglycerols from plant oil or waste biomass yields monoalky esters of long-chain fatty acids with short chain alcohol such as fatty acid methyl esters. More attention has recently focused on the application of genetically engineered microalgae in the production of biodiesel. The reaction of acetyl-CoA to malonly-CoA is catalyzed by acetyl-CoA carboxylase (ACCase). ACCase is often regarded as the first committed step of the fatty acid synthetic pathway. My aim is to increase the lipid content of the cell through enhancing the expression of ACCase. Cyanobacterium Synechocystis sp. PCC 6803 (Synechocystis) is one of the best-characterized microalgae species and, given its robust growth characteristic, becomes the organism of choice for applications. Detailed studies on techniques for genetic manipulation of this species are available. Unfortunately, no suitable vector for cyanobacteria is courrently available. Therefore, the thesis has two main aims: (1) cloning ACCase on expression vector. (2) constructing a E.coli / Synechocystis suitable shuttle vector for efficient gene cloning and functional assays. My results show that overexpression of ACCase increases the lipid content of the cells. In addition, the shuttle vector constructed was transformed into Synechocystis for testing. As it turned out, it conferred antibiotic resistance to the cell and could be recovered as a plasmid form.
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