碩士 / 國立中興大學 / 生命科學系所 / 102 / Brackish medaka (Oryzias dancena) is a famous ornamental fish in the local aquaria. Based on their transparent embryos, microinjection can be performed in the euryhaline medaka. The brush border of the enterocyte was composed of microvilli.
In mammals, absorption and secretion by intestinal epithelial cells may be facilitated by movements of the microvilli comprising the "brush border" surface of these cells.
The villin 1 is involved in the microvillus formation. Recently, villin 1-like protein(VILL), a villin 1 homologo protein, has been identified in the medaka species. The present study illustrated that the distribution of VILL protein was found in apical regions of the intestine. In addition, this study identified the upstream sequence of Odvill from the medaka genomic DNA. The Tol2 transposon-mediated system with 5'' upstream region of Odvill were used microinjection to produce transgenic embryos.
Subsequently, the enterocytes of transgenic medaka exhibited fluorescent signals of the reporter gene, AmCyan 1. And Odvill promoter/enhancer were illustrated in Odvill 5'' upstream (2.8 kb) and Intron1 (2.4 kb). In the transgenic medaka, the patterns of AmCyan 1 expression were observed in different stages embryos. These embryos exhibited fluorescent signals in the intestine at 14-16 dpf. At the same time, the larvae
were eaten. The result implied that a large numbers of microvilli would be formed during this period and performed their absorptive functions. This study was the first to develop the Tol2 transposon-mediated system for the brackish medaka. We evaluated the stronger fluorescent transgenic medaka with semi-PCR and maintained it for breeding. To date, the transgenic medaka (F1) with AmCyan1-positive fluorescence had breed for 64.7% of total transgenic medaka. Taken together, the present study applied the Tol2 transposon-mediated system for development of a new transgenic fish with fluorescent intestine.
| Identifer | oai:union.ndltd.org:TW/102NCHU5105083 |
| Date | January 2014 |
| Creators | Poa-Kai Lin, 林寶凱 |
| Contributors | 李宗翰 |
| Source Sets | National Digital Library of Theses and Dissertations in Taiwan |
| Language | zh-TW |
| Detected Language | English |
| Type | 學位論文 ; thesis |
| Format | 88 |
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