臺灣博碩士論文加值系統

本研究針對鳥類COI基因設計了11條引子，挑選其中九組適合的配對，並參考文獻發表的引子對，針對台灣地區62種100個鳥類樣品進行COI基因部分片段之PCR複製及定序，分別得到263至750 bp的部分片段序列。利用BioEdit 7.0及MEGA 4軟體進行比對分析，結果顯示大部分鳥種序列的種內歧異度皆低於種間歧異度。將所有片段分別以Kimura’s 2-parameter method計算序列間的基因距離，再以Neighbor-joining method建立各鳥種的親緣關係，結果顯示同物種的序列，在親緣關係圖上皆能被歸到同一類群。由此可知，本論文所分析的COI基因部分片段，足以作為鳥類種屬鑑定之標的。依這些片段分佈之相對位置，建立了一套系統化的鳥類種屬鑑定方法，以利刑事鑑識上之應用。此外，本論文中將定序所得的台灣鳥種COI基因序列建立資料庫，以彌補現有DNA資料庫中對於台灣地區鳥類序列資料的不足。

謝誌…………………………………………………………………I
中文摘要……………………………………………………………III
Abstract……………………………………………………………IV
目錄…………………………………………………………………V
圖目錄………………………………………………………………VII
表目錄………………………………………………………………VIII
第一章 緒論………………………………………………………1
第一節 前言……………………………………………………1
第二節 研究動機與目的………………………………………3
第三節 文獻探討………………………………………………3
一、種屬鑑定之DNA分析方法………………………………4
二、動物的粒線體DNA………………………………………8
三、Cytochrome c oxidase subunit I基因………………9
四、鳥類種屬鑑定……………………………………………12
第二章 研究方法與步驟…………………………………………14
第一節 研究流程………………………………………………14
第二節 樣品來源………………………………………………14
第三節 實驗方法………………………………………………15
一、DNA萃取…………………………………………………15
二、聚合酶連鎖反應(PCR) …………………………………16
三、PCR產物純化……………………………………………17
四、定序反應…………………………………………………18
五、序列比對…………………………………………………18
六、引子設計及引子對選擇…………………………………18
七、物種歧異度(sequence diversity)分析………………19
八、群聚分析(cluster analysis) ………………………19
九、鳥類COI序列資料庫建立………………………………19
十、盲樣試驗…………………………………………………20
第三章 結果與討論………………………………………………21
第一節 鳥類COI基因全長序列之分析………………………21
第二節 鳥類COI基因部分片段之複製及定序………………22
第三節 複製短片段COI基因通用引子對之設計……………23
第四節 短片段序列之分析…………………………………26
第五節 群聚分析……………………………………………35
一、種(Species) …………………………………………35
二、屬(Genus) ……………………………………………35
三、科(Family) …………………………………………36
四、目(Order) ……………………………………………38
第六節 序列資料庫之建立…………………………………40
第七節 盲樣試驗……………………………………………40
第四章 結論……………………………………………………42
第五章 未來展望………………………………………………43
參考文獻…………………………………………………………44
附圖………………………………………………………………51
附表………………………………………………………………64
附錄………………………………………………………………67論文全文

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