碩士 / 國立屏東科技大學 / 生物科技研究所 / 93 / Interferon, as the first line of defense in animals, plays important roles in defense against viral infections. For the purpose of understanding the molecular mechanism involved in the transcription of the interferon gene, the promoter region of the zebrafish interferon was cloned and sequenced, and its activity was further characterized using firefly luciferase transient transfection expression assay. Various fragments of the zebrafish interferon 5’-flanking region were transfected into ZFL cells, in which maximum promoter activity was found to be located in the 2.2 kb fragment of the zebrafish interferon 5’ flanking region. Influences of the viral replicative intermediate double stranded RNA by using synthetic polyinosinic -polycytidylic acid [poly(I): poly(C)] on the gene expression of interferon was also investigated in this study. All zebrafish interferon promoter fragments were treated with various doses of 1, 10, 100 μg/mL poly(I):poly(C), and the highest promoter activity was observed in the —2.2 kb under the dose of 10 μg/mL poly(I):poly(C). A series of the binding sites of putative transcription factor were derived through a computer analysis. The ontogenic gene expressions of the interferon promoter were further investigated in transgenic zebrafish, to which an interferon (2.2-kb) promoter-driven green fluorescent protein (GFP) encoding the GFP cDNA transgene was microinjected. Morphological examination of transgenic zebrafish indicated that the interferon promoter-driven GFP transcripts appeared in the lymphoid organs. These results suggested that the interferon promoter is actively expressed in a tissue-specific manner, and the interferon promoter plays an important role in virus resistance during teleost growth is therefore concluded.
| Identifer | oai:union.ndltd.org:TW/093NPUST111001 |
| Date | January 2005 |
| Creators | Yung-Kang Yu, 游原鏗 |
| Contributors | Tzou-Chi Huang, Jyh-Yih Chen, 黃 卓 治, 陳 志 毅 |
| Source Sets | National Digital Library of Theses and Dissertations in Taiwan |
| Language | zh-TW |
| Detected Language | English |
| Type | 學位論文 ; thesis |
| Format | 96 |
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