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Please use this identifier to cite or link to this item: http://ntour.ntou.edu.tw:8080/ir/handle/987654321/10620

Title: 石斑魚廣效性蛋白分解酵素抑制物之純化、分子選殖及特性分析研究
Purification, Molecular Cloning and Characterization 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: 2010-08
Issue Date: 2011-06-28T07:45:36Z
Publisher: 行政院國家科學委員會
Abstract: 摘要:石斑魚廣效性蛋白分解酵素抑制物之純化、分子選殖及特性分析研究 本研究旨在純化石斑魚血漿中抑制病原菌所分泌蛋白分解酵素 (protease) 之物 質,並做特性分析研究。將血漿樣本經FPLC 系統,以各種分離管柱進行抑制物質的純 化分離,再利用電泳分析蛋白質在自然、氧化與還原態下結構之變化,並估算其分子量, 另將進行純化物在不同生理特性下之穩定性與抑制功能;而在此抑制物質蛋白分子研究 上,將採用N 端序列定序與LC/MS/MS 對其胺基酸序列進行分析,並將定序結果與目 前已發表物種做比較。其次,選殖此抑制物質基因序列,分析比對與其他物種之差異; 而為了探討石斑魚血漿抑制物質對不同病原菌之protease 抑制能力之差異,利用養殖常 見的病原菌Vibrio alginolyticus、V. carchariae、V. harveyi、V. parahaemolyticus、Aeromonas hydrophila 以及Photobacterium damselae subsp. piscicida 之細胞外產物進行抑制試驗。最 後,利用免疫分析方法與基因選殖探討不同魚種體內抑制物質在免疫與基因特性上之差 異。 本計畫之研究期程為兩年:第一年進行石斑魚抑制細菌蛋白質分解酵素物質之蛋白 分子結構與功能性的研究;第二年選殖石斑魚抑制細菌蛋白質分解酵素物質基因,並利 用各種免疫方法與基因選殖基因序列,比較不同種石斑魚與其他物種間含抑制蛋白分解 酵素物質之差異。
Abstract:Purification, molecular cloning and characterization of the broad-spectrum protease inhibitor proteins from groupers. The present project intends to purify, molecular clone and characterize the protease inhibitors from grouper Epinephelus sp. Protease inhibitors of grouper will be purified by using various columns on Fast protein liquid chromatography system. Molecular mass and amino acid sequence of the purified protease inhibitors will be determined and compared with other references. The effects of temperature, pH value and methylamine on the activity of the purified protease inhibitors will be determined. In addition, gene sequences of the protease inhibitors will be cloned and compared with that of other animal species. The inhibitory activity of the purified protease inhibitors against protease produced by pathogenic Vibrio alginolyticus, V. carchariae, V. harveyi, V. parahaemolyticus, Aeromonas hydrophila,or Photobacterium damselae subsp. piscicida will be investigated. The immunological and genetic variations of the protease inhibitors from different fish species will be determined and compared by using various immunological and molecular cloning techniques. The project will be completed within two years. First year:The molecular structure and function of purified protease inhibitors from grouper Epinephelus sp. will be studied. Second year:Genes of the protease inhibitors will be cloned. The immunological and genetic variation of the protease inhibitors from different fish species will be determined and compared by using various immunological and molecular cloning techniques.
Relation: NSC99-2313-B019-004
URI: http://ntour.ntou.edu.tw/ir/handle/987654321/10620
Appears in Collections:[水產養殖學系] 研究計畫

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