Development of a molecular toolkit to genetically engineer the microalga Nannochloropsis gaditana CCMP526 for biotechnology applications
- Publication Type:
- Thesis
- Issue Date:
- 2020
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Model microalgae such as ๐๐ฉ๐ญ๐ข๐ฎ๐บ๐ฅ๐ฐ๐ฎ๐ฐ๐ฏ๐ข๐ด ๐ณ๐ฆ๐ช๐ฏ๐ฉ๐ข๐ณ๐ฅ๐ต๐ช๐ช and ๐๐ฉ๐ข๐ฆ๐ฐ๐ฅ๐ข๐ค๐ต๐บ๐ญ๐ถ๐ฎ ๐ต๐ณ๐ช๐ค๐ฐ๐ณ๐ฏ๐ถ๐ต๐ถ๐ฎ are well suited for genetic engineering as they possess an array of well characterised genetic tools that can be used for gene and genome manipulations. ๐๐ข๐ฏ๐ฏ๐ฐ๐ค๐ฉ๐ญ๐ฐ๐ณ๐ฐ๐ฑ๐ด๐ช๐ด ๐จ๐ข๐ฅ๐ช๐ต๐ข๐ฏ๐ข is a photosynthetic oleaginous microalga that has been studied extensively due to a broad range of industrial applications such as oil, polyunsaturated fatty acids (PUFAs) and pigments. Even though genetic engineering and synthetic biology resources are rapidly advancing and widening this algaโs industrial potential, there are still some limitations such as a narrow repertoire of functional promoters with limited diversity in terms of expression range and inducibility; and a lack of knowledge of specific regulatory elements that control transcription in ๐๐ข๐ฏ๐ฏ๐ฐ๐ค๐ฉ๐ญ๐ฐ๐ณ๐ฐ๐ฑ๐ด๐ช๐ด spp. that would allow for predictable expression and rational design of promoter regions. To date, several studies involving gene over-expression have been conducted in ๐๐ข๐ฏ๐ฏ๐ฐ๐ค๐ฉ๐ญ๐ฐ๐ณ๐ฐ๐ฑ๐ด๐ช๐ด spp. using endogenous promoters, but in-depth promoter analysis done in other model microalgae like ๐. ๐ต๐ณ๐ช๐ค๐ฐ๐ณ๐ฏ๐ถ๐ต๐ถ๐ฎ and ๐. ๐ณ๐ฆ๐ช๐ฏ๐ฉ๐ข๐ณ๐ฅ๐ต๐ช๐ช is still missing with ๐๐ข๐ฏ๐ฏ๐ฐ๐ค๐ฉ๐ญ๐ฐ๐ณ๐ฐ๐ฑ๐ด๐ช๐ด spp. By identifying and profiling a suite of promoters for use in ๐. ๐จ๐ข๐ฅ๐ช๐ต๐ข๐ฏ๐ข CCMP526, we have expanded the genetic toolbox available for this industrially relevant microalgal species both for biotechnological applications such as metabolic engineering or recombinant protein production, and to understand the biology of ๐๐ข๐ฏ๐ฏ๐ฐ๐ค๐ฉ๐ญ๐ฐ๐ณ๐ฐ๐ฑ๐ด๐ช๐ด. In this research we also explored ๐. ๐จ๐ข๐ฅ๐ช๐ต๐ข๐ฏ๐ข as a novel platform for sesquiterpenoid biosynthesis and it represents the first report of terpenoid engineering in this microalga, which shed some light into metabolic pathway engineering for terpenoid production in ๐. ๐จ๐ข๐ฅ๐ช๐ต๐ข๐ฏ๐ข. Whilst this PhD thesis offers new knowledge into terpenoid biosynthesis, it also provides candidate gene promoters, which can be used in a wide variety of biotechnology application in ๐. ๐จ๐ข๐ฅ๐ช๐ต๐ข๐ฏ๐ข.
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