利用粒腺體全基因體DNA 序列瞭解 台灣小雨蛙與近親種的演化及親源關係

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姓名

林佳賢(Jia-Sian Lin) 查詢紙本館藏

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生命科學系

論文名稱

利用粒腺體全基因體DNA 序列瞭解台灣小雨蛙與近親種的演化及親源關係
(Evolution and phylogeny of Microhyla fissipes and its relavent taxa infering from mitogenome)

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摘要(中)

小雨蛙 (Microhyla fissipes)在台灣台南(模式地點) 首次被發表命名，但並沒有被廣泛接受。本研究，作者使用從宜蘭收集的樣本代表台灣樣本，並完成完整的線粒體基因組序列（登錄號：MF673131），用以研究Microhyla fissipes 與其近親的親緣關係，特別是希望瞭解與中國樣本（NC_009422）的差異。儘管我們的樣本（16,729個鹼基對）的基因與及基因順序其他兩棲動物幾乎相同，但台灣和中國的樣本之間序列上已有了一些差異。有趣的是，我們發現ND5 和ND6 基因與其他基因演化非常不同。另外，對基因中所使用的密碼子，進行三個位元的統計分析發現，第二個位元使用胸腺嘧啶的比例較高，在第三個位元使用使用鳥嘌呤的比例明顯較低。

摘要(英)

Microhyla fissipes was first recognized in Tainan, Taiwan (type locality), however it is not highly accepted. Herein, author used the sample from Yi-Lan, Taiwan as a representative of Taiwan samples and obtained the complete mitochondrial genome sequence (accession
number: MF673131) in order to study the phylogeny of Microhyla fissipes and its relevants, especially in the comparison with the China sample (NC_009422). Although the genome of our sample (16,729 base pairs) contains genes and gene order almost identical to other amphibians’, some variations have been accumulated between Taiwan and China samples. Interestingly, we noticed that ND5 and ND6 genes evolved very differently from other genes. Also there were strongly codon usage bias with high thymine frequency at the second codon and very low guanine frequency at the third codon.

關鍵字(中)

★ 小雨蛙
★ 親緣性
★ 粒腺體基因組序列

關鍵字(英)

★ Microhyla fissipes

論文目次

中文摘要 .................................................................................................................................. ii
Abstract ........................................................................................................................................... iii
誌謝 ........................................................................................................................................... iv
目錄 .......................................................................................................................................... v
表目錄.................................................................................................................................................. vii
圖目錄.............................................................................................................................................. .. viii
縮寫檢索表.........................................................................................................................................ix
1. 緒論 .............................................................................................................................................1
1.1 小雨蛙 .....................................................................................................................................1
1.2 粒腺體.......................................................................................................................................1
1.3 關於粒腺基因體遺傳疾病.....................................................................................................3
1.4 分子生物技術的應用...............................................................................................................3
1.5 過去利用粒腺基因體對姬蛙屬(Microhyla)與狹口蛙屬(Kaloula)的研究.............................4
1.6 研究內容..................................................................................................................................5
2. 材料與方法 ...................................................................................................................................6
2.1 樣品 .........................................................................................................................................6
2.2 粗萃 .........................................................................................................................................6
2.3 引子 (primer) 設計................................................................................................................6
2.4 聚合酶連鎖反應(PCR)...........................................................................................................6
2.5 純化 .........................................................................................................................................7
2.6 定序 ...........................................................................................................................................7
2.7 整合序列資料...........................................................................................................................7
2.8 分析基因序列...........................................................................................................................7
3. 結果 ...............................................................................................................................................9
3.1 粒腺體基因...............................................................................................................................9
3.2 姬蛙屬 (Microhyla) 與狹口蛙屬(Kaloula) 基因差異.........................................................9
3.3 演化樹分析.............................................................................................................................10
3.4 密碼子現象.............................................................................................................................10
3.5 D-LOOP 與L 鏈複製起點...................................................................................................11
4. 討論 .............................................................................................................................................12
4.1 ND5 與 ND6 演化速率較特殊..........................................................................................12
4.2 台灣與中國M. fissipes 為相同物種......................................................................................12
5. 參考文獻 .....................................................................................................................................14

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指導教授

劉阜果(Fu-Guo Robert Liu)

審核日期

2018-2-21

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