碩士 / 國立臺灣海洋大學 / 水產養殖學系 / 95 / After short term of fasting, animals will regulate energy expenditure. It can enhance metabolism efficiency and strengthen animal’s livability. The effect of metabolism change by fasting in cobia（Rachycentron canadum） is not clear now. In the experiment, fasting marker genes（Asparagine synthetase and uncoupling protein 2）were cloned to improve suppression subtractive hybridization（SSH）. The improved SSH：marker-assisted suppression subtractive hybridization（MA-SSH） is developed for identifing the genes up-regulated in cobia liver after 9 days of fasting. Fasting marker genes were used for identing the quality of SSH product. It increases the chance to find the differential expression genes. In this experiment, thirty-three genes were found in SSH product. The genes can be assigned to seven major functional classes. There are carbohydrate metabolism related genes, lipid metabolism related genes, protein metabolism related genes, immune related genes, genes that respond to stress, genes that respond to detoxification and unknown function genes. These genes were quantitated by RT-PCR to observe the gene expression after 3, 6, 9 days of fasting. In thirty-three genes there were twenty genes up-regulated significantly after 9 days of fasting. Among these differential expression genes uncoupling protein 2（UCP2）and Glycerol 3-Phosphate Dehydrogenase（G3PD）were up-regulated most. The mRNA expression of UCP2 after 3, 6 and 9 days fasting was up-regulated to 16.2 fold, 6.7 fold and 4.8 fold, respectively. The mRNA expression of G3PD after 3, 6 and 9 days fasting was up-regulated to 19.2 fold, 15.2 fold and 8.3 fold, respectively. To prevent PCR bias RNA-dot blot was used to confirm the RT-PCR data of UCP2. The gene expression trend from RT-PCR is simiar to that of dot blot hybridization. It means that the RT-PCR data is credible. The other genes are up-regulated significantly.
| Identifer | oai:union.ndltd.org:TW/095NTOU5086041 |
| Date | January 2007 |
| Creators | Yau-Hung Lin, 林堯泓 |
| Contributors | Ron-Shan Cheng, 陳榮祥 |
| Source Sets | National Digital Library of Theses and Dissertations in Taiwan |
| Language | zh-TW |
| Detected Language | English |
| Type | 學位論文 ; thesis |
| Format | 83 |
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