臺灣博碩士論文加值系統

目前人類學家對於南島民族的起源有濃厚的興趣，而台灣原住民屬於南島民族的一支。因此，研究台灣原住民間的親緣關係有助於瞭解南島民族的起源問題。 人類學家早期是運用語言及文化特質來探討族群類緣，後來結合生物學的外觀性狀，如今加上遺傳學的原理來排除後天環境所造成的影響，再加上近代分子生物學的技術演進、DNA定序自動化的完備，許多遺傳指標逐漸發展成人類學研究的重要工具。目前常用的遺傳指標有人類白血球抗體基因分型、微衛星體序列、粒線體核酸序列與Y染色體單點核酸多型態等。而在眾多遺傳指標之中，HLA分型明顯受到天擇壓力影響，其它的遺傳指標是否受到天擇、是否符合中性理論，亦不無疑問。 相對於上述已知的遺傳指標，基因組中存在另一群分子序列，叫做偽基因（pedudogene）。目前認為偽基因是由基因組中具功能基因複製產生，一旦經由突變的累積而逐漸成為不具功能的片段，即成為偽基因。我們相信偽基因在天擇過程中應該是中性，但目前的文獻中，似乎還沒有以偽基因作為族群類緣的比較基礎的研究。因此我們的實驗便以偽基因作為比較基礎，用以探討台灣原住民的演化歷史。我們所選擇的偽基因是G protein-coupled receptor kinases 6 gene的processed pseudogene （ΨGRK6）。 研究結果顯示，偽基因的突變機率，比基因體平均比例要高，以NCBI網站上所記載的序列作為外群，利用最大或然率分析方法（Maximum-likelihood method）所繪製的分子分類樹可看出原住民主要分為兩大群聚，我們認為會發展成差異如此大的兩大群聚，一種較容易解釋的原因是原住民與漢族之間的通婚情形比預期普遍，但是需要更大量的樣本數來支持這個假設。 我們的結果提供了研究族群遺傳的新方向，也確立了偽基因作為比較基礎的角色，未來更可以結合其他部位（如Y染色體）的偽基因來進行研究，進一步確立偽基因的效能，更可以利用偽基因幫助釐清南島語族的起源問題。

Taiwan aborigines are part of Austronesians. Anthropologists are interested in where the origin of Austronesians was. Studying the relationships among Taiwan aborigines is important to understand the origin of Austronesians. Anthropologists have used linguistic and cultural features to study the origin and dispersal of human populations before. Physical characteristics were used later. Now, genetic markers are the new favorite tools. The genetic markers that have been used includ HLA genes, microsatellites, mitochondrial DNA sequences and Y chromosome sequences. The HLA genes are probably heavily selected throughout most of human history, and whether other genetic markers are selectively neutral is also doubtful. There has been a long dispute on where the homeland of Austronesians. Different genetic markers, pointed to different origins. None of the genetic markers used so far satisfies the premises for phylogenetic tree reconstruction. A pseudogene is derived from functional gene duplication and can not be translated into a functional protein because of the accumulation of mutations. We believe that pseudogene is selectively neutral. We used processed pseudogene-ΨGRK6 to study the phylogenetic relationship of Taiwan aborigines. Our study indicated that the mutation rate of pseudogene is much higher than expected. The DNA sequence ofΨGRK6 could be cluster into two groups. One trivial explanation of the pattern is that there was population admixture of Han Chinese and Taiwan aborigines. Our results support the proposal that pseudogene sequences are useful tools to study the dispersion of human populations.

致謝I ABSTRACTII 中文摘要III 總目錄IV 圖目錄VI 表目錄VII 序論1 1. 台灣的原住民族1 1.1台灣原住民各族介紹1 1.2台灣原住民族的起源5 2. 族群類緣關係的研究方法8 3. 偽基因-新的遺傳指標12 4. ΨGRK6介紹13 5. 研究目的14 材料與方法16 材料16 1. 血液樣本16 漢族16 原住民族16 2. 試藥與試劑組16 3. 化學藥品17 方法18 1. 自血液中抽取genomic DNA18 2. 利用聚合酵素連鎖反應放大ΨGRK6片段18 3. 偵測聚合酵素連鎖反應產物18 4. 純化聚合酵素連鎖反應產物18 5. ΨGRK6的選殖19 6. ΨGRK6的DNA定序19 7. 序列分析20 結果21 1. ΨGRK6片段聚合酵素連鎖反應結果21 2. 以限制酵素ECORⅠ偵測ΨGRK6的選殖21 3. ΨGRK6的DNA定序21 4. 原住民各族與漢族的變異點22 5. 分子演化樹22 討論24 參考資料30 附錄48 附錄一、台灣南島語言分佈圖48 附錄二、世界南島語族分佈圖49

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