碩士 / 國立臺灣海洋大學 / 水產養殖學系 / 94 / Cobia is a new species with high economical value and potential international market. However, high percentage of fish oil is added in the formulated feeds to meet the requirement of polyunsaturated fatty acid for cobia becomes a barrier for the development of cobia culture. In addition to use vegetable oil to substitute fish oil partly, researches on pathways for synthesis of polyunsaturated fatty acid and genes encoding enzymes catalyzing each step are necessary. Therefore, 1) a cobia fin cell line is established without bacteria, fungi and mycoplasma contamination. The cell line is cultured in L-15 medium containing 10 % FBS at 28 oC and foreign genes can be transferred into cells using BTX 830 electroporator using 4 mm cuvette containing PBS at condition of one pulse of 450 voltage. Cells can be cryopreserved in FBS containing 10% DMSO. 2) Promoters, cDNA, 5’ and 3’ UTR sequences of delta 6 desaturase and elongase genes are cloned. Both promoters contain sterol regulatory elements. The cDNA sequences and amino acid sequence of both genes are highly conserved among fishes. Variable amounts of both genes mRNA are detected in tissues tests but highest amounts of both genes mRNA are detected in brain. 3) The plasmids containing either delta 6 desaturase or elongase cDNA are delivered into cobia fin cell line and recombinant proteins are expressed. Most of fusion proteins of delta 6 desaturase and green fluorescent protein are located at specific location in cytoplasm. The dimers of elongase recombinant protein are detected using Western blot. As regard to the biological function of these recombinant proteins remain to be evaluated.
| Identifer | oai:union.ndltd.org:TW/094NTOU5086005 |
| Date | January 2006 |
| Creators | Yu-Jun Wang, 王鈺君 |
| Contributors | Jau-Der Chen, Ta-Chih Cheng, 陳昭德, 鄭達智 |
| Source Sets | National Digital Library of Theses and Dissertations in Taiwan |
| Language | zh-TW |
| Detected Language | English |
| Type | 學位論文 ; thesis |
| Format | 95 |
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