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| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.author | Ya-Wen Gan | en |
| dc.contributor.author | 甘雅文 | zh_TW |
| dc.date.accessioned | 2021-07-01T08:12:53Z | - |
| dc.date.available | 2021-07-01T08:12:53Z | - |
| dc.date.issued | 2003 | |
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| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/75372 | - |
| dc.description.abstract | 地錢(Marchantia polymorpha L.)是一種具有異型性染色體的蘚類，其配子體因而為雌雄異株。在地錢雌雄配子體中的性染色體則分別被定名為X及Y染色體。前人針對地錢的基因體研究發現，在其Y染色體所特有的重複性序列中，存在著一個可轉錄的基因ORF162，並且證明其轉錄產物只在雄配子枝中被偵測到。然而由本研究的結果發現，台灣所產的同種地錢中，其雌配子體之基因體也具有ORF162的同源基因；而由進一步的分析結果來看，這個同源基因很可能是位在X染色體上。經由PCR、DNA分子選殖以及序列比對分析，本研究共在台灣產地錢的雄配子體中鑑定出兩個不同的份數(copies)的ORF162類似序列，分別命名為ORF162-MPT1和ORF162-MPT2；而在台灣產地錢的雌配子體中則鑑定出十二個不同的份數，分別命名為ORF162-MPT3、ORF162-MPT4、ORF162-MPT5、ORF162-MPT6、 ORF162-MPT7、ORF162-MPT8、ORF162-MPT9、ORF162-MPT10、ORF162-MPT11、ORF162-MPT12、ORF162-MPT13、ORF162-MPT14，前三者鑑定自雌配子體的基因體DNA，後九者鑑定自其cDNA。在分析比對DNA的啟動子序列之後來，發現它們和先前鑑定之ORF162的啟動子有顯著的不同，包含了14?27個取代或插入/移除之核?酸。而在開啟讀碼框(open reading frame)中，亦有4?59個核?酸的差異。從RT-PCR的結果顯示，ORF162-MPT1/2在雄配子枝發育的各個階段均有表現，而ORF162-MPT6/7/8/9/10/11/12/13/14則在雌配子枝發育初期有微量轉錄產物。經由cDNA的定序結果顯示, ORF162-MPT6/7/8/9/10/11/12/13/14的cDNA中各含有數個終止密碼(stop codons)，因此我們認為該轉錄產物應不會轉譯出蛋白質，同時ORF162-MPT3/5的開啟讀碼框(open reading frame)序列中亦有數個終止密碼,是故ORT162-MPT3/5/6/7/8/9/10/11/12/13/14應為假基因(pseudogohes)。此外，雖然ORF162-MPT4具有完整的開啟讀碼框，但在本研究中並未偵測到其表現，因此有待進一步的研究其是否為具有功能的基因。 | zh_TW |
| dc.description.abstract | (前文內容丟失)also several stop codons in the sequences of 0RF162-MPT3/5, thus 0RF162-MPT3/5/6/7/8/9/1O/11/12/13/14 are all likely to be pseudogenes. The only exception is ORFl62-MPT4, which has an identical open reading frame with 0RF162-MPT1/2, but no expression was detected in our analysis, and awaits for further studies. | en |
| dc.description.provenance | Made available in DSpace on 2021-07-01T08:12:53Z (GMT). No. of bitstreams: 0 Previous issue date: 2003 | en |
| dc.description.tableofcontents | 中文摘要…………………………………………………i 英文摘要…………………………………………………ii 第一章 前言…………………………………………………1 第二章 材料與方法…………………………………………………6 1. 材料採集與無菌培養…………………………………………………6 1.1 苞芽無菌培養…………………………………………………6 1.2 土壤培養…………………………………………………6 2. Genomic DNA的萃取…………………………………………………6 3. ORF162-MPT的分子選殖…………………………………………………7 3.1 聚合?連鎖反應(PCR)…………………………………………………8 3.2 PCR產物的分子選殖(cloning)…………………………………………………8 3.2.1 少量質體DNA的抽取…………………………………………………8 3.2.2 勝任細胞(competent cell)的製備…………………………………………………9 3.2.3 接合反應(Ligation)…………………………………………………11 3.2.4 轉形作用(Transformation)…………………………………………………11 3.2.5 質體快速檢定(Quick screening)…………………………………………………11 4. 性別專一性引子設計與PCR放大…………………………………………………12 5. DNA dot blot…………………………………………………13 6. 南方轉漬分析(Southern blot analysis)…………………………………………………14 6.1 非放射性標定探針的製備…………………………………………………14 6.2 Genomic DNA的限制?作用及轉漬…………………………………………………15 6.3 雜合反應(hybridization)與訊息偵測…………………………………………………16 7. 啟動子序列的分子選殖…………………………………………………18 8. 總量RNA之抽取…………………………………………………19 8.1 Pine Tree Method…………………………………………………19 8.2 Collcert Plant RNA Reagent…………………………………………………20 9. RT-PCR…………………………………………………21 9.1 First-Strand cDNA Synthesis…………………………………………………21 9.2 Second-Strand cDNA Synthesis…………………………………………………21 10.PCR產物的分子選殖…………………………………………………22 10.1 接合反應…………………………………………………22 10.2 轉形作用…………………………………………………23 10.3 Conoly PCR…………………………………………………23 11. 核酸及胺基酸序列排序的應用軟體…………………………………………………24 第三章 結果…………………………………………………25 1. 性別專一性引子測試結果…………………………………………………25 2. 地錢雌雄株基因體內ORF162-MPT之份數…………………………………………………25 3. 序列分析…………………………………………………26 4. ORF162-MPT在地錢雌雄株之表現…………………………………………………28 5. 其他蘚類DNA dot blot之結果…………………………………………………29 第四章 討論…………………………………………………30 1. 性別專一性引子測試…………………………………………………30 2. ORF162-MPT的所在位置…………………………………………………30 3. 雌基因體中ORF162-MPT的起源…………………………………………………31 4. ORF162-MPT的份數…………………………………………………32 5. ORF162-MPT之角色…………………………………………………33 引用文獻…………………………………………………36 表…………………………………………………41 圖…………………………………………………49 附錄…………………………………………………93 | |
| dc.language.iso | zh-TW | |
| dc.title | 台灣?地錢之雌雄性染色體上的假設性基因ORF162-MPT之研究 | zh_TW |
| dc.title | Identification and characterization of ORF162-MPT, a putative gene on the male and female sex chromosomes,from Marchantia polymorpha L.in Taiwan | en |
| dc.date.schoolyear | 91-2 | |
| dc.description.degree | 碩士 | |
| dc.relation.page | 104 | |
| dc.rights.note | 未授權 | |
| dc.contributor.author-dept | 生命科學院 | zh_TW |
| dc.contributor.author-dept | 植物科學研究所 | zh_TW |
| 顯示於系所單位： | 植物科學研究所 | |
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