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Please use this identifier to cite or link to this item: http://ntour.ntou.edu.tw:8080/ir/handle/987654321/30558

Title: 石斑魚廣效性蛋白分解酵素抑制物分子選殖與序列分析
Molecular Cloning and Sequence Analysis of the Broad-Spectrum Protease Inhibitor Proteins from Groupers
Authors: 李國誥
Contributors: NTOU:Department of Aquaculture
國立臺灣海洋大學:水產養殖學系
Date: 2011-08
Issue Date: 2012-04-13T01:03:32Z
Publisher: 行政院國家科學委員會
Abstract: 摘要:本研究旨在由石斑魚肝臟組織中選殖蛋白分解酵素抑制物-巨球蛋白之基因序列, 以進行序列比對與功能性序列特性分析,且利用基因體生物資訊方法針對石斑魚與台灣 常見的淡海水養殖魚種找尋其關聯性;其次,藉由純化之蛋白質抑制物獲取抗血清以免 疫分析方法-交叉免疫電泳、火箭電泳、蛋白質斑點印漬分析 (Dot-Blot Assay) 以及 Western Blot等方法,針對台灣常見的數種養殖與野生石斑魚以及淡海水養殖魚種進行免 疫分析,探討不同魚種血液之間所含蛋白分解酵素抑制物的差異變化。最後,利用即時 定量PCR (real-time PCR) 分析石斑魚遭受細菌性蛋白分解酵素攻擊後,其體內蛋白酵素 抑制物之基因表現量,藉以了解蛋白酵素抑制物在石斑魚免疫防禦上所扮演之角色。
Abstract:The present project intends to clone the gene sequence of alpha-2-macroglobulin (the protease inhibitor) from liver of grouper for sequence comparison and function characterization. The relationship of alpha-2-macroglobulin gene among major fish cultured in Taiwan will be studied using bioinformatic technologies. The activities of alpha-2-macroglobulin in cultured fish and wild groupers will also be determined by immunological methods i.e. CIE, rocket IE, Dot-blot assay and Western blot, using rabbit antiserum against purified alpha-2-macroglobulin. The gene expression of alpha-2-macroglobulin in groupers after challenge with bacterial protease will be determined using real-time PCR technique to understand its possible role playing in the defense mechanism.
Relation: NSC100-2313-B019-008
URI: http://ntour.ntou.edu.tw/handle/987654321/30558
Appears in Collections:[水產養殖學系] 研究計畫

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