碩士 / 國立臺灣大學 / 漁業科學研究所 / 89 / Two classes of tilapia c-ski cDNA (Accession number: AJ012011, AJ012012), designated as tski1 and tski2, respectively encoded a 687 and a 714 AA protein and shared a 57% AA identity. Comparison with the Ski proteins of chickens, humans and Xenopus, tilapia TSki polypeptides shared a 60, 57 and 57% (TSki1) and 67, 63 and 61% (TSki2) AA identity, respectively. The tski1 and tski2 gene extend a minimum of 16 kb and 20kb in the tilapia genome. The exon 1 comprising 1.5 kb (tski1) and 1.3kb (tski2) of cDNA sequence have been mapped with screening tilopia genomic DNA library. Then exons 1 and 2 are separated by more than 10 kb (tski1) and 2kb (tski2) of genomic sequence. Using SSP-PCR (single specific primer polymerase chain reaction) and PCR with genomic DNA templates, it is clear that tski1 and tski2 cDNA are encoded seven exons. In this two genes, the exons 2 through 7 are closely linked within 2 kb in the tilopia genome. The long 3 untranslated region is contiguous in the genome with the distal portion of the open reading frame such that the exon 7 is composed of both coding and noncoding sequence.
The most and the least abundant c-ski mRNAs are located in the brain and the skeletal muscle, respectively. Both tski1 and tski2 were widely expressed in the adult tissues examined, but tski2 transcripts were at higher levels except in the ovary and oocytes: tski1 transcripts were predominant in the ovary, whereas tski2 transcripts were predominant in the testes. In the oocytes, the tski1 mRNA was a maternally-inherited stockpile that subsequently was degraded, so that the expression ratio of tski1 to tski2 transcripts declined gradually as the fish developed from oocyte to 4-cm fry.
The notochord, eye, muscle fiber, tail fin and heart transient expression pattern showed that the 3 kb tski1 promoter promotes tski1 gene wildly in zebrafish. As to the pT2EGFP, had the same EGFP expression pattern with pT1EGFP but weak and mosaic extremely.
| Identifer | oai:union.ndltd.org:TW/089NTU00451016 |
| Date | January 2001 |
| Creators | Chiu-Ju Huang, 黃秋茹 |
| Contributors | Huai-Jen Tsai, 蔡懷楨 |
| Source Sets | National Digital Library of Theses and Dissertations in Taiwan |
| Language | zh-TW |
| Detected Language | English |
| Type | 學位論文 ; thesis |
| Format | 56 |
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