臺灣博碩士論文加值系統

中文摘要
M8836013
論文名稱：應用AFLP於香魚片中有毒河魨之檢測 頁數：86
校所組別：國立屏東科技大學食品科學系研究所碩士班
畢業時間及摘要別：九十學年度第二學期碩士學位論文摘要
研究生:丁慶成 指導教授: 許祥純、廖遠東博士
論文摘要內容：
河魨為具有神經毒素之魚類，誤食其毒素會造成中毒及死亡，目前河魨之檢測方法僅能以生物檢測法測定毒性，但無法判別出是何種有毒之河魨，因此本研究針對河魨進行品種鑑別，萃取13種河魨及河魨香魚片之DNA，加入限制酶進行切割反應，再以PCR增幅限制片段，利用擴大片段長度多型性(AFLP)分析，建立有效的AFLP指紋圖譜及其實驗條件，用以快速鑑別有毒及無毒品種。結果發現限制酶切割及轉接子連接分別為37℃, 2小時及20℃隔夜反應較為適當，而預擴大反應條件則為94℃ 30 秒、56℃ 1分鐘、72℃ 1分鐘共20循環。擴大反應則以touch down 的方式設定annealing 溫度每循環降0.7℃(65→56℃)共12循環，之後再進行23循環的擴大反應，條件為94℃ 30秒、56℃ 30秒、72℃ 1分鐘共23循環。使用的九種引子組合，以A2、B1、B2、B3、C1的檢測效果最佳，可檢測出河魨品種間的差異性，且可發現在同屬中有毒與無毒河魨具有多型性片段，進而辨別河魨品種。
本研究亦使用20組Operon kit primer 做RAPD增幅，發現最適DNA濃度為200ng ，增幅條件94℃ 2分鐘、94℃ 1分鐘、37℃ 2分鐘、72℃ 2分鐘，連續30個循環再以94℃ 1分鐘、37℃ 2分鐘、72℃ 12分鐘進行PCR反應，結果OPAA.6、OPAA.9、OPAA.10、OPAA.14、OPZ.5、OPZ.6、OPY.4、OPY.6、OPY.7等九組引子具有較佳增幅結果，也可以應用於河魨種源鑑別之參考。

目錄
頁次
中文摘要…………………………………………………….……. I
英文摘要………………………………...……………..…………III
誌謝………………………………………….…………………….V
目錄………………………………...……………………………..VI
圖索引…………………………………………………………….IX
表索引…………………………………....………………………Xll
第一章 前言…………………...…………………………………1
第二章 文獻回顧………….……………..………………………4
2.1食用河魨之歷史………………………...……………………..4
2.2河魨分類…………………………………...…………………..5
2.2.1魨形目(Tetrodontiformes)……………..…………………..6
2.2.2鱗魨亞目(Balistidei)…………………...………………….6
2.2.3箱魨亞目(Ostracioidei)………….……..………………….6
2.2.4魨亞目(Tetrodontoidei)………….…….….……………….6
2.3河魨魚獲狀況…………….………….…….…………………..7
2.4河魨之加工利用與管理…….……….…….…………………..9
2.5河魨毒(tetrodotoxin, TTX)……………….…………………..13
2.5.1中毒作用機轉…………….……………………………...13
2.5.2河魨毒之鑑定……………………………………………19
2.5.2.1薄層分析…………………………………………….19
2.5.2.2電泳分析…………………………………………….19
2.5.2.3高效能液相層析…………………………………….19
2.6蛋白質分析………………………………………….………..20
2.7 DNA比對…………………………………….……….………21
2.7.1 PCR技術………………………………….…….………..22
2.7.2限制酵素片段長度多型性………….……….…………..22
2.7.3 RAPD-PCR……………………………..………………..25
2.7.4 AFLP…………………………………………..…………26
2.7.4.1轉接子之黏接(ligation of adapters)…….……..…….26
2.7.4.2 PCR增幅(PCR amplification)………….…………...27
2.8分析方法………………………………………….……….….27
2.9 AFLP應用………………………………………..………..….28
第三章 材料與方法………………………………..…………...31
3.1實驗架構………………………………………..…………….31
3.2樣品來源………………………………………..…………….32
3.3.1生肉………………………………………..……………..32
3.3.2香魚片製備………………………..……………………..32
3.4 DNA萃取……………………………..………………………34
3.4.1.儀器……………………………………..………………..34
3.4.2.試劑……………………………………..………………..34
3.4.3. DNA萃取…………………………….…………….……35
3.4.4. DNA濃度及品質鑑定………………….……….………35
3.5 AFLP分析…………………………….……..……….……….36
3.5.1 DNA之限制酵素切割反應………………..…….………36
3.5.2轉接子接合……………………………...……………….36
3.5.3預擴大反應………………………...…………………….37
3.5.4選擇性擴大多型性片段反應…………….…...…………37
3.5.5電泳…………………………………...………………….38
3.5.6銀染……………………...…………………………….39
3.6 RAPD-PCR…………………...…………..………………..…39
3.6.1 PCR反應液…………………..……..…………………...40
3.6.2 PCR增幅條件……………………..…….……………....40
3.6.3 PCR增幅產物分析……………..……………………….40
3.7圖譜分析………………………………….….……………….40
第四章 結果與討論………………………….……….………...42
4.1 AFLP…………………………………………..……………...42
4.1.1 AFLP操作條件之建立……………….……….………...42
4.1.1.1限制酵素切割及轉接子接合之條件建立……….…42
4.1.1.2選擇性預擴大反應條件之建立………………...…..43
4.1.1.3選擇性擴大反應條件之建立…………………...…..43
4.1.2 AFLP圖譜分析…………………………………………..44
4.2 RAPD…………………………………………………………55
4.2.1不同河魨DNA濃度對RAPD產物之影響………….…55
4.2.2新鮮河魨及加工香魚片不同之RAPD增幅片段………55
4.2.3不同引子組RAPD增幅片段之比較…………..……..…57
第五章 總結………………………………………………...…..67
第六章 參考文獻……………………………...………………..72
作者簡介……………………………………………………...…..86

第六章.參考文獻
1. 方素嫣 (2000) 梅子種源之快速鑑別與在加工上之利用。臺灣大學園藝學碩士論文15-30。
2. 王育恕 (1997) 利用cDNA-AFLP技術進行甘藷中受甲基化/去甲基化調控及塊根發育相關基因之研究。國立台灣大學 碩士論文 13-39。
3. 朱玉灼、鄭森雄 (1987) 以電泳方法判別河魨與安康魚及澳洲與越南冷凍紅狗母魚之差異。中國水產 321:3-6。
4. 杜先覺、傅幼敏、施養志、張碧秋、周薰修、呂雅蕙、黃登福 (1999) 香魚片引起食物中毒之首例鑑定報告。藥物食品分析 7(2):163-167。
5. 沈世傑 (1993) 台灣魚類圖鑑。台灣大學動物系 台北 台灣 506-818。
6. 林頎生、徐仁鏤、簡秀玲 (1992) 市售香魚片衛生狀況及蛋白特色判定之可行性1-8。
7. 邵廣昭 (1996) 臺灣常見魚介貝類圖說。臺灣省魚業局 台北 台灣249-255。
8. 洪聖儀、蔡麗琴、賴淑雅、李俊億 (1999) 限制性複製片段長度多型性在動物種屬鑑定之研究。刑事科學 47:45-65。
9. 范廷佳 (1998) 圓斑河魨轉錄因子STAT基因之選殖與表現 。國立台灣大學碩士論文 11-58。
10. 孫寶年、李國誥、翁秀貞 (1986) 台灣地區常見食用魚貝類圖說。行政院衛生署 台北 台灣 64-65。
11. 徐克學 ( 1996 ) 數量分類學。台北 台灣104-122。
12. 徐泰浩、曾耀銘 (1995) 生物技術概論。藝軒圖書出版社 台北 台灣33-48。
13. 常抗美、施維德 (2000) 江浙河魨資源、養殖和消費調查。科學養魚 8:7-8。
14. 張春霖、成慶泰、鄭葆珊、李思忠、鄭文蓮、王文濱 (1994) 黃渤海魚類。水產出版社 台北 台灣 309-332。
15. 陳孟仙 (1984) 臺灣海域產鯖河魨屬魚類、分類、生殖及食性之研究。臺灣大學海洋研究所碩士論文15-30。
16. 陳孟仙、張崑雄、李信徹 (1986) 由肌肉蛋白之比較探討臺灣產鯖河魨屬魚類之系統分類。中央研究院動物集刊 67-77。
17. 陳源高、陳亞芬、劉正文 (1999) 暗紋東方魨人工繁殖中幾個關鍵技術。科學養魚 9:17-18。
18. 彭紹楠、劉傳正 (1984) 解凍河魨肌肉之毒素轉移。臺灣省水產試驗所 382:17-23。
19. 黃登福 (1985)海洋生物毒。中國水產 386:4-17。
20. 黃登福 (1987) 河魨毒化學機構之探討。中國水產 413:31-37。
21. 黃登福 (1990) 日本之河魨加工利用、管理與養殖。中國水產 452:7-17。
22. 黃登福 (1994) 海洋生物毒引起之魚貝類食物中毒及其相關研究。生命科學簡訊 8：2-9。
23. 黃登福 (1999) 台灣因海洋生物毒河組織胺引起之魚貝介類食物中毒及其相關研究。食品工業 31(6)：19-30。
24. 黃登福 (1999) 海洋生物毒和生理活性物質。中醫藥雜誌 10(1):23-50。
25. 黃登福 (1999) 認識河魨香魚片。海大漁推 28：1-30。
26. 黃登福、邵廣昭 (1995) 台灣魚市場有毒魚貝類圖鑑。台灣省漁業局出版 台北 台灣 239-250。
27. 黃登福、高麗玲、鄭森雄 (1989) 台灣市販香魚片(調味魚乾)之化學特性與毒性研究。食品科學 16 (4)：425-434。
28. 黃登福、張雯如、饒國峰、鄭森雄 (1994) 以河魨製成香魚片(調味魚乾)之加工研究。食品科學 21(1)：58-66。
29. 林麗娟 (1991) 臺灣產織紋螺類Nassariidae之毒性及其毒成份。國立海洋大學碩士論文7-11。
30. 楊正義 (1982) 澎湖河魨乾加工。豐年 32(14):37-38。
31. 葉蕙玲、陳茂松 (1982) 臺灣產河魨毒性初步調查。Bulletin of Taiwan Fisheries Research Institute 34:337-340。
32. 潘宏彬、袁金球 (1999) 河魨燒尾病和貓頭蚤併發症的防治。科學養魚 10:29。
33. 蔡永祥、黃登福 (2000) 拼死吃河魨-談河魨中毒。漁業推廣 165：13-27。
34. 蕭泉原、闕銘洲、莊健隆 ( 1982) 臺灣河魨利用現況。中國水產 351:10-13。
35. 賴文雅 (1998) 利用AFLP比對分析系統分析文心蘭栽培種之遺傳圖譜。台灣大學碩士論文 1-66。
36. 錢建興 (1999) 河魨魚的越冬與管理。科學養魚 11:33。
37. 謝幸媚、吳坤龍、李俊億 (2000) 植物DNA鑑定在犯罪偵查上之應用與評估。刑事科學(49):1-7。
38. 謝喻文 (1999) 利用細胞色素b基因之PCR技術及基因序列作為紅鰭多紀魨與橫紋多紀魨原料及其加工品之魚種鑑定.台灣海洋大學碩士論文 1-66。
39. Ajmone-Marsan, P., Valentini, A., Cassandro, M., Vecchiotti-Anntaldi, G. B. and Kuiper, M. (1997) AFLPTM markers for DNA fingerprinting in cattle. Animal Genetics. 28:418-426.
40. Appa Rao, K. B. C., Bhat, K. V. and Totey, S. M. (1996) Detection of species-specific genetic markers in farm animals through random amplified polymorphic DNA (RAPD) Genetic Analysis: Biomolecular Engineering. 13:135-138.
41. Asensio, L., Gonzalez, I., Fernandez, A., Rodriguez, MA., Lobo, E., Hernandez, P.E., Garcia, T. and Martin, R. (2002) Application of random amplified polymorphic DNA (RAPD) analysis for identification of grouper (Epinephelus guaza), wreck fish (Polyprion americanus), and Nile perch (Lates niloticus) fillets. Food Prot. 65(2):432-5.
42. Bai, G., Ayele, M., Tefera, H. and Nguyen, H.T. (1999) Amplified fragment length polymorphism analysis of Tef [Eragrostis tef (Zucc.)Trotter]. Crop Sci. 39:819-824.
43. Barker, J. H. A., Matthes, M., Arnold, M., Edwards, K. J., Ahman, I., Larsson, S. and Karp, A. (1999) Characterisation of genetic diversity in potential biomass willows (Salix spp.) by RAPD and AFLP analyses. Genome. 42:173-183.
44. Bertini, L., Potenza, L., Zambonelli, A. and Amicucci, A. (1998) Restriction fragment length polymorphism species-specific patterns in the identification of white truffles. FEMS Micro. Letters 164:397-401.
45. Birstein, V.J. and DeSalle, R. (1998) Molecular phylogeny of Acipenserinae. Mol. Phylogenet Evol. 1:141-55.
46. Botstein, D., White, R. L., Skolnick, M. H. and Davies, R. W. (1980) Construction of genetic linkage map in man using restriction fragment length polymorphisms. Am. J. Hum. Genet. 32:314-331.
47. Bremer, J. R. 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. Fisheries Sci. 127:507-511.
48. Brunner, B., Todt, T., Lenzner, S., Stout, K., Schulz, U., Ropers, H., and Kalscheuer, V., (1999) Genomic structure and comparative analysis of nine Fugu gene: Conservation of synteny with Human chromosome Xp22.2-p22.1. Genome research 5:437-450.
49. Chen, X., Lim, S. H., Wong, S. M., Lee, Y. H., Kuo, J., Yam, T. W. and Lin, J. J. (1999) Amplified fragment length polymorphism analysis of vandaceous orchids. Plant Science 141:183-189.
50. Chikuni, K., Ozutsumi, K., Koishikawa, T. and Kato, S. (1990) Species identification of cooked meats by DNA hybridization assay. Meat Sci. 27:119-128.
51. Chikuni, K., Tabata, T., Kosugiyama, M., and Monma, M. (1994) Polymerase chain reaction assay for detection of sheep and goat meats. Meat Sci. 37:337-345.
52. Chou, S. S., Tsai, Y. H., Chan, P. and Hwang, D. F. (1997) Tetrodotoxin-associated food poisoning due to ingesting fish. J.of food and drug analysis 2(1):77-81.
53. Ebbehoj, K. F. and Thomsen, P. D. (1991) Species differentiation of heated meat products by DNA hybridization. Meat Sci. 30: 221-234.
54. Edwards, YJ., Elgar, G., Clark, MS. and Bishop, MJ. (1998) The identification and characterization of microsatellites in the compact genome of the Japanese pufferfish, Fugu rubripes: perspectives in functional and comparative genomic analyses. Mol. Biol. 278(4):843-854.
55. Fairbrother, K. S., Hopwood, A. J., Lockley, A. K. and Bardsley, R. G. (1998) Meat speciation by restriction fragment length polymorphism analysis using an α-actin cDNA probe. Meat Sci. 50(1):105-114.
56. Finnila, S., Hassinen, I. and Majamaa, K. (1999) Restriction fragment analysis as fragment length polymorphism markers. Aquatic Botany. 58: 165-172.
57. Finnila, S., Hassinen, I. And Majamaa, K. (1999) Restriction fragment analysis as a source of errer in detection of heteroplasmic mtDNA mutations. Mutation research genomics 406:109-114.
58. Goodyear, P. D., MacLaughlin-Black, S. and Mason, I. J. (1994) Purification of highly degraded DNA by gel filtration for PCR. Bio. Tech. 16:235-238.
59. Guoli, Z., Mingguang, Z., Zhijiang, Z., Hongsheng, O., and Qiang, L. (1999) Establishment and application of a polymerase chain reaction for the identification of beef. Meat Sci. 51: 233-236.
60. Happ, G., Aquilla, E., Martick, M., Yuncker, C., Wojciechowski, J. and Fox, l. (1999) DLA-DRB1 histocompatibility genotyping using RT-nested PCR and cycle sequencing. Vet Immunol & Immunopathol 69:93-100.
61. Herbergs, J., Siwek, M., Crooijmans, RPMA., POEL, JJ van der Poel.,and Groenen man. (1999) Multicolour fluorescent detection and mapping of AFLP markers in chicken (Gallus domesticus). Animal genetics 30: 274-285.
62. Hookey, J. V., Edwards, V., Patel, S., Richardson, J. F. and Cookson, B. D. (1999) Use of fluorescent amplified fragment length polymorphism (fAFLP) to characterize methicillin-resistant Staphylococcus aureus. J. of Microbiol Methods 37:7-15.
63. Hopwood, A. J., Fairbrother, K. S., Lockley, A. K and Bardsley, R. G. (1999) An actin gene-related polymerase chain reaction (PCR) test for identification of chicken in meat mixtures. Meat Sci. 53: 227-231.
64. Hsieh, Y. P., Woodward, B. B. and Ho, S. H. (1995) Detection of species substitution in raw and cooked meats using immunoassays. J. Food Prot. 58(5): 555-559.
65. Hunt, D. J., Parkes, H. C. and Lumley, I. D (1997) Identification of the species of origin of raw cooked meat products using oligonucleotide probe. Food Chem. 60(3):437-442.
66. Huys, G. and Swings, J. (1999) Evaluation of a fluorescent amplified fragment length polymorphism (AFLP) methodology for the genotypic discrimination of Aeromonas taxa. FEMS microbiology letters 177:83-92.
67. Hyytia, E., Hielm, S., Bjorkroth, J. and Korkeala, H. (1999) Biodiversity of Clostridium botulinum type E strains isolated from fish and fishery products. Appl. Environ. Microbiol. 65(5): 2057-2064.
68. Jones, J. T. and Harrower, B. E. (1998) A comparison of the efficiency of differential display and cDNA-AFLPs as tools for the isolation of differentially expressed parasite genes. Fundam. Appl. Nematol. 21(1):81-88.
69. Kim, S.H., Han, A., Kronatad, J. and Breuil, C. (1999) Differentiation of sapstain fungi by restriction fragment length polymorphism patterns in nuclear small subunit ribosomal DNA. FEMS Microbiology Letters 177:151-157.
70. Knorr, C., Cheng, H. H. and Dodgson, J. B. (1999) Application of AFLP markers to genome mapping in poultry. Animal Genetics 30:28-35.
71. Ko, JM., Do, GS., Suh, DY., Seo, BB., Shin, DC. and Moon, HP. (2002) Identification and chromosomal organization of two rye genome-specific RAPD products useful as introgression markers in wheat. Genome 45(1):157-164.
72. Kobayashi, T., Okuzumi, M. and Fujii, T. (1995) Microflora of fermented puffer fish ovaries in rice-bran “Fugunoko Nukazuke”. Fisheries Sci. 6(2):291-295.
73. Koch, G and Jung, .C (1997) Phylogenetic relationships of industrial chicory varieties revealed by RAPDs and AFLPs. Agronomie. 17: 323-333.
74. Koh, M. C., Lim, C. H., Chus, S. B., Chew, S. T., and Phang, S. T. W. (1998) Random amplified polymorphic DNA (RAPD) fingerprints for identification of red meat animal species. Meat Sci. 48(3/4): 275-285.
75. Lawyer, FC., Stoffel, S., Saiki, RK., Chang, SY., Landre, PA., Abramson, RD. and Gelfand, DH. (1993) High-level expression, purification and enzymatic characterization of full-length Thermus aquaticus DNA polymerase and a truncated form deficient in 5'' to 3'' exonuclease activity. PCR Methods 2(4):275-87.
76. Lee, J. C. and Chang, J. G. (1994) Random amplified polymorphic DNA polymerase chain reaction (RAPD-PCR) fingerprints in forensic species identification. Forensic Sci. 67: 103-107.
77. Lehours, P., Rogues, AM., Occhialini, A., Boulestreau, H., Gachie, JP. and Megraud, F., (2002) Investigation of an outbreak due to Alcaligenes xylosoxydans subspecies xylosoxydans by random amplified polymorphic DNA analysis. Eur. J. Clin. Microbiol Infect Dis. 21(2):108-113.
78. Li, W-H. (1981) Simple method for constructing phylogenetic tree from distance matrices. Proceeding national academic society USA. 78(2): 1095-1089.
79. Liu, A., Nichols, P. and Dunham, R. A. (1998) Inheritance and usefulness of AFLP markers in channel catfish, blue catfish and their F1, F2, and backcross hybrids. Mol. Gen. Genet. 258:260-268.
80. Liu, Z. J., Li, P., Argue, B. J. and Dunham, R. A. (1999) Random amplified polymophic DNA markers: usefulness for gene mapping and analysis of genetic variation of catfish. Aquaculture 174:59-68.
81. Martinez, I. and Yman, I. M. (1998) Species identification in meat products by RAPD analysis. Food Res. Intern. 31(6): 459-466.
82. Matsui, T., Yamamori, K., Furukawa, K. and Kono, M. (2000) Purification and some properties of a tetrodotoxin binding protein from the blood plasma of kusafugu, Takifugu niphobles. Toxicon 38:463-468.
83. Matsumura, K. (1998) Production of tetrodotoxin in puffer fish embryos Env. Toxicol & Pharmacol 6:217-219.
84. Matsunaga, T., Chikuni, K., Tanabe, R., Muroya, S., Shibata, K., Yamada, J. and Shinmura, Y. (1999) A quick and simple method for the identification of meat species and meat products by PCR assay. Meat Sci. 51:143-148.
85. Miyamoto. T., Okabe. H. Z., Trevanich. T., Asoh. K., Tomoda. K., Honjoh. S. and Hatano. S. (1997) Application of RAPD DNA analysis for Salmonella spp in food. J. Food Prot. 61(7): 784-791.
86. Nei, M. and li, W. H. (1979) Mathematical model for studying genetic variation in terms of restriction endonuclease. Proceeding national academic society. USA. 76(10):5269-5273.
87. Papadopouios, S., Benter, T., Anastassiou, G., Pape, M., Gerhard, S., Bomfeld, N., Ludwig, WD. and Dorken, B.(2002) Assessment of genomic instability in breast cancer and uveal melanoma by random amplified polymorphic DNA analysis. Int. J. Cancer 99(2):193-200.
88. Parani, M., Rao, R., Mathan, N., Mathan, N., Anuratha, C., Narayanan, K. and Parida, A. (1997) Molecular phylogeny of mangroves III parentage analysis of a Phizophora hybrid using random amplified polymorphic DNA and restriction fragment length polymorphism markers. Aquatic Botany 58: 165-172.
89. Partis, L., Croan, D., Guo, Z., Clark, R., Coldham, T. and Murby, J. (2000) Evaluation of a DNA fingerprinting method for determining the species origin of meats. Meat Sci. 54:699-705.
90. Paul, S., Wachira, F. N., Powell, W. and Waugh, R. (1997) Diversity and genetic differentiation among populations of Indian and Kenyan tea (Camellia sinensis (L.) O. Kuntze) revealed by AFLP markers. Theor. Appl. Genet. 94:255-263.
91. Qi, X. and Lindhout, P. (1997) Development of AFLP markers in barley. Mol. Gen. Gent. 254: 330-336.
92. Quinteiro, J., Sotelo, C. G., Rehbein, H., Piyde, S. E., Medina, I., Perez-Martin, R. I., Rey-Mendez, M. and Mackie, I. M. (1998) Use of mtDNA direct polymerase chain reaction (PCR) sequencing and PCR-restriction fragment length polymorphism methodologies in species identification of canned tuna. J. Agric. Food Chem. 46:1662-1669.
93. Rosendahl, S., Jensen, AB., Gargas, A. and Eilenberg, J. (1998) Relationships of the insect-pathogenic order Entomophthorales (Zygomycota, Fungi) based on phylogenetic analyses of nuclear small subunit ribosomal DNA sequences (SSU rDNA). Fungal Genet Biol. 24(3):325-34.
94. Saiki, RK., Walsh, PS., Levenson, CH. and Erlich, HA.(1989)
Genetic analysis of amplified DNA with immobilized sequence-specific oligonucleotide probes. Proc. Natl. Acad. Sci. USA. 86(16):6230-4.
95. Santis, R. and Azzi, A. (2000) Use of denaturing gradient gel electrophoresis for human polymavirus JC sequence analysis. J. of Virol. Methods 85:101-108.
96. Sato, S., Ogata, T., Borja, V., Gonzales, C., Fukuyo, Y. and Kodama, M. (2000) Frequent occurrence of paralytic shellfish poisoning toxins as dominant toxins in marine puffer from tropical Toxicon 38:1101-1109.
97. Tokai, T., Sato, R. and Kitahara, T. (1995) Short-term forecasting of landings of ocellate puffer takifugu rubripes migratin around a spawning area in the inland sea of Japan. Fisheries Sci. 6(3):428-433.
98. Vos, P., Hogers, R., Bleeker, M., Reijans, M., Lee, T. V. D., Hornes, M., Frijters, A., Pot, J., Peleman, J., Kuiper, M. and Zabeau, M. (1995) AFLP：a new technique for DNA fingerprinting. Necleic Acids Res. 23(21):4407-4414.
99. Wang, C. H. and Smith, D. M. (1995) Lactate dehydrogenase monoclonal antibody immunoassay for detection of turkey meat in beef and pork. J.food sci. 60(2):253-256.
100. Weng YJ. and Chen DM. (2000) Molecular markers and its clone for salt tolerance gene in wheat. Yi. Chuan Xue. Bao. 29(4):343-9.
101. Williams, J. G. K., Kubelik, A. R., Livak, K. J., Rafalski, J. A. and Tingy, S. V. (1990) DNA polymorphisms amplified by arbitrary primers are useful as genetic markers. Nucleic Acids Res. 18:6531-6535.
102. Wolf, C., Hibner, P. and Lithy, J. (1999) Differentiation of sturgeon species by PCR-RFLP. Food Research Inter. 32:699-705.
103. Wolf, C., Rentsch, J. and Hubner, P. (1999). PCR-RFLP analysis of mitochondrial DNA: a reliable method for species identification. J. Agric. Food Chem. 47: 1350-1355.
104. Wu, Y., Delgado, R., Costello, R., Sunderland, T., Dukoff, R. and Csako, G. (2000) Quantitative assessment of aplipoprotein E gentypes by image analysis of PCR-RFLP fragments. Clinica Chimica Acta. 293:213-221.
105. Yamada, J. and Shinmura, Y. (1999) A quick and simple method for the identification of meat species abd meat products by PCR assay. Meat Sci. 51:143-148.
106. Yamashita, M. Y., Yamagishi, Y. and Yasumoto, T. (1995) 5,6,11 Trideoxyterodotoxin the puffer fish, fugu poecilonotus. Tetrahedron Letters 36(51):9329-9332.
107. Yoda, T., Morita, T., Kawatsu, K., Sueki, K., Shibata, T. and Hamano, Y. (1995) Cloning and sequencing of the chaperonin-encoding Cctd gene from Fugu rubripes rubripes. Gene 166(2):249-53.
108. Zhou, J. and Guo, S. (1999) Molecular typing of staphylococcus aureus with randomly amplified polymorphic DNA. Hunan Yi Ke Da Xue Xue Bao. 24(1):17-9.