臺灣博碩士論文加值系統

飛魚全世界有 8 屬 52 種，而本省有 4 屬 20 種。近年來，飛魚季等觀光活動的盛行，使得飛魚的漁獲量有明顯的減少的趨勢。而在東北部的漁民，會利用編織草蓆捕獲魚卵，雖可提高飛魚的經濟價值，但卻也因此造成飛魚的生態浩劫。漁業署統計在 2003 年國內飛魚產量為 460 公噸，而在 2007 年國內飛魚產量明顯的減少為 71 公噸，顯示飛魚日漸枯竭的現象，飛魚資源的維護已成為重要工作。
因此本研究採用兩種蛋白質電泳技術 SDS-PAGE (Sodium dodecyl sulfate-polyacrylamide gel electrophoresis) 以及 Urea-IEF (Urea- isoelectric focusing electrophoresis) 分析台灣常見的五種飛魚肌肉蛋白質組成，以探討這些電泳技術應用於鑑定生鮮飛魚魚種之可行性。並嘗試將此兩種電泳技術應用於鑑定飛魚魚卵的種類。五種飛魚魚肉在 8M Urea、水溶性及鹽溶性等蛋白質萃取後之 SDS-PAGE 圖譜，其差異性僅蛋白質亮帶表現量的強弱，以及微量的種特異性蛋白存在，其鑑定效果十分有限。但 SDS-PAGE 圖譜在鑑定五種飛魚的魚卵上，可以有效地鑑定出白鰭飛魚及斑鰭飛魚的種類。
在 Urea-IEF 的部分，在魚肉蛋白質的部分，其鹽溶性蛋白質可有效鑑定出五種飛魚魚種。在魚卵的部分，鹽溶性蛋白質亦為鑑定出五種飛魚魚卵種類的最佳方法。由此可知利用 SDS-PAGE 與 Urea-IEF 的圖譜結果，可以藉由鹽溶性蛋白鑑定出此市售魚卵的種類為白鰭飛魚。

There are 8 genus and 52 species of flying fish in the world. Nowadays, there are lots of traveling activity within flying fishes seasons, it attack flying fish harvest.Taiwan fisheries Agency report showed harvest of flying fish in 2003 was 460 tones, however harvest in 2007 decreased to 71 tones. Hence, flying fish preservation is becoming an important issue.
This research uses two protein electrophoresis, SDS-PAGE (Sodium dodecyl sulfate-polyacrylamide gel electrophoresis) and Urea-IEF (Urea- isoelectric focusing electrophoresis) to analyze 5 flying fish species in taiwan. Furthermore, use these electrophoresis to identify roes of flying fish species. Resaults show 8M Urea-extracted proteins, sarcoplasmic proteins and myofibrillar proteins can not display obvious differential protein band in SDS-PAGE. However, SDS-PAGE can display roes of flying fish from Cypselurus unicolor and Cypselurus poecilopterus.
The part of Urea-IEF show myofibrillar proteins can identify 5 flying fishes species, it is the best identifying technique in this study. Moreover, for roe of flying fish, salt-soluble proteins can identify 5 flying fishes species. Summarize all of results from SDS-PAGE and Urea-IEF,we can identify commercial roe of flying fish as Cypselurus unicolor by using salt-soluble proteins.

壹、前言 4
貳、文獻整理 7
一、飛魚的特性、經濟、利用及其相關文化之重要性 7
(一) 地域分類與生物特徵 7
1. 黑鰭飛魚 8
2. 白鰭飛魚 9
3. 紫斑鰭飛魚 9
4. 紅斑鰭飛魚 10
5. 斑鰭飛魚 10
(二) 漁業經濟 11
(三) 相關利用 12
(四) 台灣各地飛魚相關活動及其文化 12
1. 蘭嶼達悟族飛魚文化 12
2. 墾丁飛魚季 16
3. 綠島飛魚季 17
4. 台東、花蓮阿美族飛魚季 17
5. 基隆、宜蘭之飛魚卵漁業 18
二、常見鑑定魚種技術之介紹 19
(一) 以外觀特徵鑑定魚種 19
(二) 以基因分析法鑑定魚種 20
1. 直接定序法 20
2. 聚合酶連鎖反應-隨機放大多型性 DNA 21
3. 聚合酶連鎖反應-限制片段長度多型性 21
4. 擴增片段長度多型性分析技術 22
5. 多重引子-聚合酶連鎖反應 22
(三) 以蛋白質分析法鑑定魚種 22
1. 十二烷基硫酸鈉聚丙烯醯胺膠體電泳 23
2. 等電點焦聚電泳法 24
3. 二維電泳法 25
4. 毛細管電泳法 26
三、魚肉肌肉萃取蛋白質與魚種鑑定之關聯性 27
(一) 肌漿蛋白 27
(二) 肌原纖維蛋白 28
1. 肌凝蛋白 28
2. 原肌凝蛋白 29
參、研究內容 30
第一章、 利用 SDS-PAGE 電泳技術鑑定五種台灣常見飛魚及魚卵之魚種
一、前言 30
二、實驗材料 31
三、實驗方法 33
四、結果 38
五、討論 42
六、圖表 44
第二章、 利用 Urea-IEF 電泳技術鑑定五種台灣常見飛魚及魚卵之魚種
一、前言 63
二、實驗材料 64
三、實驗方法 65
四、結果 68
五、討論 71
六、圖表 73
肆、綜合結論 88
伍、參考文獻 89
謝辭 100

王侯。(1983)。飛魚與柴魚。漁友月刊 4：35-36。
尤心正。(1999)。飛魚卵脂質成分及其低溫貯藏中之變化。國立台灣海洋大學食品科學系碩士論文。
田哲益 (達西烏拉彎．畢馬)。(2002)。台灣的原住民-達悟族，台原出版社。
沈世傑。(1993)。台灣魚類誌。國立台灣大學動物學系印行， 188-195。
余光弘。(1998)。臺灣原住民史雅美族篇。中央研究院民族學研究所。
邵廣昭。(1996)。台灣常見魚介貝類圖說 (下)-魚類。台灣省漁業局印行，55。
林宜鉦。(1959)。談飛魚。中國水產月刊 47：13。
洪建德。(1999)。海面飛躍的梭-飛烏。漁業推廣 155：7。
洪國堯。(2008)。建立飛魚漁業管理制度之成果與展望。農政與農情，第 194 期。
哈帽。(2000)。東海岸豐濱的飛魚季節來臨了。台灣原住民月刊電子報 (5)。
孫瑞茂。(2002)。飛魚乾加工法之研究。國立屏東科技大學食品科學系碩士學位論文。
徐瀛洲。(1990)。蘭嶼之美。行政院文化建設委員會印行，26-35。
陳南宏。(2007)。笛鯛科魚類之毒性分析及其魚種蛋白質電泳技術鑑定方法之探討。國立台灣海洋大學食品學系碩士論文。
陳春輝。(1978)。台灣近海飛魚分類。中國文化學院海洋研究所碩士論文。
陳萱瑜。(2007)。台灣原住民族文化活動之研究以蘭嶼達悟族飛魚祭為例。私立銘傳大學設計管理研究所碩士論文。
黃韻潔。(2011)。利用魚肉蛋白電泳技術鑑定鯙鰻魚種之研究。國立台灣海洋大學食品科學系博士論文。
張燦穩。(1991)。蘭嶼椰油村雅美族人漁場利用型態。國立台灣師範大學地理研究所碩士學位論文。
達屴。(2001)。東海岸阿美族的燻飛魚乾。台灣原住民月刊電子報 (19)。
楊碧月。(2002)。蘭嶼飛魚季-飛魚的故鄉。社團法人台灣環境資訊協會訊電子報。
劉振鄉。(1997)。深具觀光潛力的蘭嶼飛魚文化。漁業推廣， 125：34-36。
漁業署。(2003)。台灣地區漁業年報。農委會漁業署印行。
漁業署。(2007)。台灣地區漁業年報。農委會漁業署印行。
衛惠林、劉斌雄。(1962)。蘭嶼雅美族的社會組織。中央研究院民族學研究所專刊，113-126。
蔡敬廷。(2005)。飛魚組胺酸相關化合物與生物胺形成之探討。國立台灣海洋大學食品科學系碩士學位論文。
闕銘洲。(2004)。飛魚乾製品品質與化學特性及其貯藏期間變化。國立台灣海洋大學食品科學系碩士學位論文。
An, H., Wei, C. I., Zhao, J., Marshall, M. R. and Lee, C. M. (1989). Electrophoretic identification of fish species used in surimi products. Journal of Food Science, 54: 253-257.
Asghar, A., Samejima, K. and Yasui, T. (1984). Functionality of muscle protein in gelation mechanism of structured meat products. CRC Critical Reviews in Food Science and Nutrition, 22: 27-43.
Beismann, H., Barker, H., Karp, A. and Speck, T. (1997). AFLP analysissheds light on distribution of two Salix species and their hybrid alonga natural gradient. Molecular Ecology, 6: 989-993.
Berkes, F. (1987). The common property resource problem and the fisheries off Barbados and Jamaica. Environmental Management, 36: 540-545.
Bollag, D. M., Rozycki, D. M. and Edelstein, S. J. (1996). Isoelectric focusing and two dimensional gel electrophoresis.Protein Methods, 2nd. Ed by Rozyki, D. M. and Edelstein, S. J., Wiley-Liss, New York. 173-193.
Broughton, R. E. and Dowling, T. M. (1994). Length variation in mitochondrial DNA of the minnow Cyprinella spilopterus. Genetics, 138: 179-190.
Chen, C. S., Hwang, D. C. and Jiang, S. T. (1988). Purification and characterization of milkfish myosin. Nippon Suisan Gakkaishi, 54: 1423-1427.
Chen, T. Y. and Hwang, D. F. (2002). Electrophoretic identification of muscle proteins in 7 puffer species. Journal of Food Science, 67: 936-942.
Chen, T. Y., Shiau, C. Y., Noguchi, T., Wei, C. I. and Hwang, D. F. (2003). Identification of puffer fish species by native isoelectric focusing technique. Food Chemistry, 83: 475-479.
Chen, T. Y., Shiau, C. Y., Wei, C. I. and Hwang, D. F. (2004). Preliminary study on puffer fish proteome-species identification of puffer fish by two-dimensional electrophoresis. Journal of Agricultural and Food Chemistry, 52: 2236-2241.
Chiou, S. H. and Wu, S. H. (1999). Evaluation of commonly used electrophoretic methods for the analysis of proteins and peptides and their application to biotechnology. Analytica Chimica Acta, 383: 47-60.
Chou, S. S., Su, S. C., Shiau, H. W., Hwang, D. F., Yu, P. C. and Lee, S. C. (1998). Comparison of protein and amino profiles in natural and artificial shark's fins by using capillary electrophoresis. Journal of Food Science, 63: 782-784.
Comesana, A. S., Abella, P. and Sanjuan, A. (2003). Molecular identification of five commercial flatfish species by PCR - RFLP analysis of a 12S RNA gene fragment. Journal Science Food Agriculture, 83: 752-759.
Dalzell, P. (1993). The fisheries biology of flying fishes (Families: Exocoetidae and Hemiramphidae) from the Camotes Sea, Central Philippines. Journal of Fisheries Science and Technology, 43: 19-32.
Etienne, M., Jerome, M., Fleurence, J., Rehbein, H., Kundiger, R., Yman, I. M., Ferm, M., Craig, A., Mackie, I. M., Jessen, F., Smelt, A. and Luten, J. (1999). A standardized method of identification of raw and heat-processed fish by urea isoelectric focusing: A collaborative study. Electrophoresis, 20: 1923-1933.
Gallardo, J. M., Sotelo, C. G., Pineiro, C. and Perez Martin, R. I. (1995). Use of capillary zone electrophoresis for fish species identification. Differentiation of flat fish species. Journal of Agricultural and Food Chemistry, 43: 1238-1244.
Hashimoto, K., Watabe, S., Kono, M. and Shiro, K. (1979). Muscle protein composition of sardine andmackerel. Bulletin of the Japanese Society of Scientific Fisheries, 45: 1435-1441.
Hashimoto, K., Watabe, S., Nakagawa, T. and Sorita, K. (1984). Electrophoretic identification of three subspecies of the genus Lagocephalus pufferfish. Bulletin of the Japanese Society of Scientific Fisheries, 50: 115-118.
Huang, T. S., Chen, J. S., Marshall, M. R. and Wei, C. I. (1990). Quantification of shrimp in shrimp-surimi mixtures using urea gel isoelectric focusing. Journal of Food Science, 55: 1206-1209.
Hsieh, Y. W., Hwang, P. A., Pan, H. H., Chen, J. B. and Hwang, D. F. (2003). Identification of tetrodotoxin and fish species in an adulterated dried mullet roe implicated in food poisoning. Journal of Food science, 68: 142-146.
Hwang, D. F., Hsieh, Y. W., Shiu, Y. C, Chen, S. K. and Cheng, C. A. (2002). Identification of tetrodotoxin and fish species in a dried dressed fish fillet implicated in food poisoning. Journal of Food Protection, 65: 389-392.
Jaime, R., Bremer, A., Zhang, L., Bulak, J. S. and Ely, B. (1998). A polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) assay for the discrimination of mitochondrial DNA from the Florida and northern subspecies of largemouth bass. American Fisheries Society, 127: 507-511.
Jerome, M., Lemaire, C.,Bautista, J. M., Fleurence, J. and Etienne, M. (2003). Molecular phylogeny and species identification of sardines. Journal of Agricultural and Food Chemistry, 51: 43-50.
Karaiskou, N., Apostolidis, A. P., Triantafyllidis, A., Kouvatsi, A. and Triantaphyllidis, C. (2003). Genetic identification and phylogeny of thee species of the genus Trachurus based on mitochondrial DNA analysis. Marine Biotechnology, 5: 493-504.
Khatchaturov, V. C. (1983). The digestive system of the flying fishes (Exocoetidae, Beloniformes). Journal of Applied Ichthyology, 23: 106-113.
Knuutinen, J. and Harjula, P. (1998). Identification of fish species by reversed-phase high-performance liquid chromatography with photodiode-array detection. Journal of Chromatography B, 705: 11-21.
Laemmli, U. K. (1970). Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature, 227: 680-685.
Larrain, M. A., Abugoch, L., Quitral, V., Vinagre, J. and Segovia, C. (2002). Capillary zone electrophoresis as a method for identification of golden kinglip (Genypterus blacodes) species during frozen storage. Food Chemistry, 76: 377-384.
Longhurst, A. R. and Pauly, D. (1987). Ecology of Tropical Oceans. London: Academic Press.
Lundstrom, R. C. (1980). Fish species identification by thin layer polyacryamide gel isoelectric focusing: Collaborative study. Journal of Association of Official Analytical Chemists, 63: 69-73.
Mackie, I. M. (1990). Identifying species of fish. Analytical Proceedings, 27: 89-92.
Mackie, I. M. (1997). Methods of identifying species of raw and processed fish. Fish Processing Technology. 2nd Ed by Hall, G. M., Blackie Academic and Professional, London, 160-199.
Mackie, I. M., Craig, A., Etienne, M., Jerome, M., Fleurence, J., Jessen, F., Smelt, A., Kruijt, A., Yman, M. I., Ferm, M., Martinez, I., Martin, R. P., Pineiro, C., Rehbein, H. and Kundiger, R. (2000). Species identification of smoked and gravad fish products by sodium dodecylsulphate polyacrylamide gel electrophoresis, urea isoelectric focusing and native isoelectric focusing: A collaborative study. Food Chemistry, 71: 1-7.
Mackie, I. M., Pryde, S. E., Gonzales-Sotelo, C., Medina, I., Perez - Martin, R., Quinteiro, J., Rey-Mendez, M. and Rehbein, H. (1999). Challenges in the identification of species of canned fish. Trends in Food Science and Technology, 10: 9-14.
Matsuura, M. and Arai, K. (1987). Preparation properties of myosin native thick filaments from fish dorsal muscle. Nippon Suisan Gakkaishi, 53: 1073-1082.
Nakagawa, T., Watabe, S. and Hashimoto, K. (1988a). Electrophoretic analysis of sarcoplasmic proteins from fish muscle on polyacrylamide gels. Nippon Suisan Gakkaishi, 54: 993-998.
Nakagawa, T., Watabe, S. and Hashimoto, K. (1988b). Identification of three major components in fish sarcoplasmic proteins. Nippon Suisan Gakkaishi, 54: 999-1004.
Niwa, E., Suzuki, M. and Nakayama, T. (1985). Heat-induced polymerization of fish actomyosin. Bulletin of the Japanese Society of Scientific Fisheries, 51: 1795-1798.
Ojima, T. and Nishita, K. (1987). Dissociation of regulatory light chains from hybrid myosin. Nippon Suisan Gakkaishi, 53: 93-98.
Ojima, T. (1989). Troponin and tropomyosin. Comparative Biochemistry of Muscular Protein in Aquatic Animals Ed by Arai, K., Koseisai-Koseigaku, Tokyo, 51-61.
Pineiro, C., Barros-Velazquez, J., Sotelo, C. G., and Gallardo, J. M. (1999). The use of two-dimensional electrophoresis in the characterization of the water-soluble protein fraction of commercial flat fish species. Zeitschrift fur Lebensmittel-Untersuchung und-Forschung A, 208: 342-348.
Plowman, J. E. and Herbert, B. R. (1992). Identification of the species of origin of cooked fish by isoelectric focusing. LWT-Food Science and Technology, 25: 224-227.
Potts, A. C., Thomas, A. D. and Nichols, E. (2003). An economic and social assessment of the flying fish (pelagic) fishery of Tobago, Trinidad and Toba. Proc. Gulf and Caribbean Fisheries Institute, 54: 635-649.
Rao, K. S. and Basheeruddin, S. (1973). Unusual catches of the flying fish, Parexocoetus brachypterus (Richardson) in inshore waters at Madras. Indian Journal of Fish, 20: 629-634.
Rehbein, H. (1990). Electrophoretic techniques for species identification of fishery products. Zeitschrift fur Lebensmittel-Untersuchung und-Forschung, 191: 1-10.
Rehbein, H., Etienne, M., Jerome, M., Hattula, T., Knudsen, L. B., Jessen, F., Luten, J. B., Bouquet, W., Mackie, I. M., Ritchie, A. H., Martin, R. and Mendes, R. (1995). Influence of variation in methodology on the reliability of the isoelectric focusing method of fish species identification. Food Chemistry, 52: 193-197.
Righetti, P. G., Bossi, A. and Gelfi, C. (1998). Title. In：Gel Electrophoresis of Proteins: A Paractical Approach. 3rd ed.(Ed. by Hames, B, D, and Rickwood, D.), IRL Press, Oxford. pp. 13-33, 127-187.
Rosen, D. E. (1984). Zeiform as primitive plectognath fishes. American Museum Novitates, 2782: 1-45.
Ross, C., Tilghman, R. W., Hartmann, J. X. and Mari, F. (1997). Distribution of parvalbumin isotypes in adult snook and their potential applications as species-specific biomarkers. Journal of Food Biochemistry, 51: 561-572.
Scobbie, A. E. and Mackie, I. M. (1988). The use of sodium dodecyl sulphate polyacrylamide gel electrophoresis in fish species identification-A procedure suitable for cooked and raw fish. Journal of the Science of Food and Agriculture, 44: 343-351.
Sebastio, P., Zanelli, P. and Neri, T. M. (2001). Identification of anchovy (Engraulis encrasicholus L.) and gilt sardine (Sardinella aurita) by polymerase chain reaction, sequence of their mitocgondrial cytochrome b gene, and restriction analysis of polymerase chain reaction products in semipreserves. Journal of Food Science, 49: 1194-1199.
Sotelo, C. G., Pineiro, C., Gallardo, J. M. and Perez-Martin, R. I. (1993). Fish species identification in seafood products. Trends in Food Science and Technology, 4: 395-401.
Steiner, J. J., Poklemba, C. J., Fjellstrom, R. G. and Elliott, L. F. (1995). A rapid one-tube genomic DNA extraction process for PCR and RAPD analysis. Nucleic Acids Research, 23: 2569-2570.
Stepien, C. A., Dixon, M. T. and Hillis, D. M. (1993). Evolutionary relationships of the Blennioid fishes families Clinidae, Labrisomidae and Chaenopsidae: Congruence between DNA sequence and allozyme data. Bulletin of Marine Science, 52: 496-515.
Suzuki, T. (1981). Fish and Krill Proteins: Processing Technology. Applied Science Publishers Ltd., London.
Tagliaro, F., Manetto, G., Crivellente, F. and Smith, F. P. (1998). A brief introduction to capillary electrophoresis. Forensic Science International, 92: 75-88.
Takeyama, H., Chow, S., Tsuzuki, H. and Matunage, T. (2001). Mitochondrrial DNA sequence variation with and between tuna Thunnus species and its application to species identification. Journal of Fish Biology, 58: 1646-1657.
Tyler, J. C. (1980). Osteology, phylogeny, and higher classification of the fishes of the order Plectognathi (Tetraodontiformes). NOAA Technical Report NMFS Circular, 434: 1-422.
Wei, C. I., An, H., Chen, J. S. and Marshall, M. R. (1990). Use of a modified urea gel isoelectric focusing method for species identification of raw or boiled white, pink, and rock shrimp. Journal of Food Biochemistry, 14: 91-102.
WinterBottom, R. (1970). Evolution of the pelvic fin ray musculature of some Triacanthoid fishes (Plectognathi). Copeia, 3: 453-456.
Yowell, K. and Flurkey, W. (1986). Effect of freezing and microwave heating on proteins from codfish fillets: Analysis by SDS polyacrylamide gel electrophoresis. Journal of Food Science, 51: 508-509.