碩士 / 國立成功大學 / 生物科技研究所碩博士班 / 93 / Groupers is a major mariculture in Taiwan and other Southeast Asian nowadays. We cloned the myostatin cDNA (1,599bp) from the orange-spotted grouper (Epinephelus coioides), and found that its deduced protein sequence is highly conserved among the mammalian and other fish species. Myostatin, a member of the transforming growth factor-β superfamily, is a negative regulator of skeletal muscle growth. Animals with inactivating mutations of myostatin have marked muscle hypertrophy, which is also called double- muscling phenotype.
We use real-time quantitative RT-PCR method to quantify the expression level of myostatin in various tissues. In grouper, myostatin mRNA is expressed mainly in muscle and eye. On the other hand, the expression of myostatin seems to increase lightly in the muscle of NNV-infected groupers in contrast with healthy ones. The phenomenon of myostatin increase is possible related to the effect of NNV infection.
Ten putative muscle-specific E-boxes exist in the promoter of grouper we cloned. And GFP expression of a promoter-reporter construct can be detected in transfected grouper cell line (GF-1). A series of deletion is also tested in GF-1, and such elimination of important cis-elements, such as TATA box, results in reduced reporter gene expression.
An application of immuno-depletion technique, we produce a peptide containing several repeats of the potential epitopes . And this antigen will have the higher antigenicity to increase the immune responds. Finally, we can inject the fusion antigens into groupers and make it grow faster and stronger by blocking the function of endogenous myostatin. And higher growth rate and antiserum titer was observed. Therefore, the higher product amount of fish meat will make the time to market shorter, and can lower the risk during mariculture.
| Identifer | oai:union.ndltd.org:TW/093NCKU5111005 |
| Date | January 2005 |
| Creators | Chih-Yueh Wei, 魏志嶽 |
| Contributors | Tzong-Yueh Chen, Huey-Lang Yang, 陳宗嶽, 楊惠郎 |
| Source Sets | National Digital Library of Theses and Dissertations in Taiwan |
| Language | zh-TW |
| Detected Language | English |
| Type | 學位論文 ; thesis |
| Format | 68 |
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