碩士 / 國立臺灣海洋大學 / 生命科學暨生物科技學系 / 106 / Diatom is a major primary producer in the marine ecosystem. It is also responsible for a part of the global carbon cycle. Its abundant unsaturated fatty acids have the potential to develop commercial products. It is a kind of eukaryotic unicellular algae which is very important for technical applications on marine ecology and biotechnology. Most of the past research has focused on the related basic fields of algae ecology and environmental science. In recent years, attention has been paid to the development of diatoms in the biotechnology field, while the research on genetic tools such as multiple gene knockdowns are still in progress. This study intends to identify the Phaeodactylum tricornutum U6 promoter and apply it to the development of diatom gene knockdown technology to increase the oil content. In this study, we used the U6 promoter sequence to establish the gene transfer vector which drive the specific shRNA and found that shRNAs were efficiently driven by this U6 promoter in the gene silencing vector. The expression of exogenous enhanced green fluorescence protein (egfp) gene was interfered with the introduction of the gene silencing vector. The highest gene inhibition rate was about 90%. The advantages of our gene silencing system include simultaneously regulating multiple target genes. Currently, we are designing a vector to drive specific shRNAs which target to genes related to metabolic pathways in diatoms, so that diatoms can produce more valuable metabolites. For example, we design a specific shRNA of the genes for two different oil-producing pathways such as phosphoenolpyruvate carboxykinase (PEPCK) and nitrate reductase (NR), the U6 promoter in the same plasmid drives the silence of the target genes, so as to achieve the purpose of promoting the oil content in diatoms.
| Identifer | oai:union.ndltd.org:TW/106NTOU5613016 |
| Date | January 2018 |
| Creators | Lin, Yi-Ling, 林羿伶 |
| Contributors | Lin, Han-Jia, 林翰佳 |
| Source Sets | National Digital Library of Theses and Dissertations in Taiwan |
| Language | zh-TW |
| Detected Language | English |
| Type | 學位論文 ; thesis |
| Format | 50 |
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