臺灣博碩士論文加值系統

豬隻具有極高的經濟價值，不僅容易成為竊盜之標的，更甚者為兇殺命案之爭端，加上近年來黑心豬肉層出不窮，因此需要一套有效的豬隻身分鑑定方法。本研究針對Sus scrofa兩個亞種，包含12個品系，合計104個隨機個體豬隻之粒線體DNA D-loop區進行分析，除設計出複製D-loop區全長之引子外，亦設計出短片段序列引子組合以提高裂解嚴重檢體之複製成功率。而經由D-loop區域定序分析後發現，豬隻D-loop區被其內一段VNTR序列（Variable number tandem repeat）劃分為三個區塊。其中，VNTR前區域有31個SNP變異位點，可區分成17種類型；VNTR區域有9種VNTR重複次數；VNTR後區域有12個SNP變異位點及3種序列插入情形，可區分成14種類型；綜合以上，共可將104個隨機個體區分成53種單倍型。粒線體DNA D-loop區個化之鑑別力（Pd）為：D-loop全區鑑別力值為0.966、VNTR前為0.889、VNTR為0.782、VNTR後為0.8。另亦針對母子組合樣品共9組18隻進行D-loop區域分析，發現母子樣品在鹼基變異位置、類型及VNTR重複次數均得到一致的結果，確認粒線體DNA的母系遺傳特性。本研究所建立之系統除可有效協助警方偵辦豬隻失竊或黑心豬肉案件外，對於建立種豬身分資料庫亦具有相當的參考價值。

謝誌 I
中文摘要 III
Abstract IV
目錄 V
表目錄 VII
圖目錄 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 5
一、我國常見豬隻品系與分類 5
二、粒線體與粒線體DNA 9
三、粒線體DNA D-loop區 10
四、豬隻粒線體DNA D-loop區相關之研究 17
第二章 研究材料與方法 22
第一節 研究流程 22
一、D-loop區序列之變異分析 22
二、D-loop區VNTR之型別分析 22
三、D-loop區序列分析系統的建立 22
第二節 樣品來源 24
ㄧ、隨機獨立個體樣品 24
二、親子組合樣品 25
三、GenBank搜尋之資料 25
第三節 實驗方法 26
一、DNA萃取 26
二、聚合酶連鎖反應（PCR） 27
三、PCR產物純化 28
四、定序反應 29
五、序列比對 29
六、引子設計及引子對選擇 29
第四節 統計方法 30
一、相符機率（Probability of matching=Pm） 30
二、鑑別力Pd值（Probability of discrimination） 30
三、序列的差異性（sequence diversity或稱genetic diversity） 30
第三章 結果與討論 31
第一節 複製D-LOOP區全長序列之引子設計 31
第二節 豬隻粒線體DNA D-LOOP區序列分析 35
一、各分類階層序列變異及單倍型頻率分析 38
二、鹼基變異情形分析 64
三、VNTR區域分析 69
四、D-loop區序列資料統計分析 71
五、本研究與台灣地區先前其他研究之比較 74
第三節D-LOOP區短片段序列之引子設計 76
第四節 具親子血緣之樣品分析 79
第四章 結論 81
第五章 未來展望 83
第六章 參考文獻 84

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