碩士 / 明道大學 / 材料科學與工程學系碩士班 / 97 / Establishment and characterization of cobia (Rachycentron canadum) cell lines derived from cobia brain (CB) is described. Brain from juvenile cobia were dissociated for 5 min, in phosphate buffered saline containing 0.25% trypsin at 25°C. The optimal culture condition for dissociated cells (primary cell culture) was at 28°C in Leibovitz's-15 medium containing 10% fetal bovine serum. The cells have been sub-cultured at a ratio of 1:2 for more than 160 passages in a period of 3 years. Origin of the cultured cells was identified by comparison of their sequences of mitochondrial cytochrome oxidase subunit I genes (cox I) with the cox 1 sequence from cobia muscle tissue. Results of the sequence confirmed brain cell line came from cobia. The cell line also shows polyploidy chromosome and no mycoplasma contamination was detected. Susceptibility to grouper iridovirus was observed on brain cell line. Meanwhile brain cell line expressed green fluorescent protein after being transfected with green fluorescent reporter gene driven by cytomegarlovirus promoter and cobia beta actin 3 promoter. The established cobia brain cell line will be valuable for pathogenesis and functional gene studies.
| Identifer | oai:union.ndltd.org:TW/097MDU05159017 |
| Date | January 2009 |
| Creators | I-Yuan Lin, 林義淵 |
| Contributors | Ta-Chih Cheng, 鄭達智 |
| Source Sets | National Digital Library of Theses and Dissertations in Taiwan |
| Language | zh-TW |
| Detected Language | English |
| Type | 學位論文 ; thesis |
| Format | 78 |
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