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| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 孫錦虹(Chin-Hung Sun) | |
| dc.contributor.author | Jo-Yu Liao | en |
| dc.contributor.author | 廖若妤 | zh_TW |
| dc.date.accessioned | 2021-06-08T01:13:49Z | - |
| dc.date.copyright | 2014-10-09 | |
| dc.date.issued | 2014 | |
| dc.date.submitted | 2014-08-13 | |
| dc.identifier.citation | Adam, R. D. (2001). Biology of Giardia lamblia. Clin Microbiol Rev, 14(3), 447-475.
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| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/18595 | - |
| dc.description.abstract | 梨形鞭毛蟲屬於人畜共通的單細胞真核寄生蟲，造成的疾病為giardiasis，他會造成腹瀉、腹痛，嚴重時可能會導致營養吸收不良的情況，梨形鞭毛蟲主要分成兩個型態-滋養體和囊體，在宿主體內的時候主要是以滋養體的型態寄生，也是造成giardiasis病徵的原因，當梨形鞭毛蟲經由宿主腸道排出體外後會以囊體的型態在外界生存，不管是在滋養體的雙核或是囊體的四核，對於有關於梨形鞭毛蟲的遺傳物質的任何機轉都是需要精細且準確地進行，當遺傳物質DNA在進行像是複製、轉錄、重組、染色體的濃縮或分離時會造成拓樸學結構上的問題，因此會需要特殊的蛋白質-拓樸異構酶來幫助遺傳物質解決結構上問題。我們經由序列的分析比對找出梨形鞭毛蟲中屬於topoisomerase type IA的topoisomerase Ⅲalpha(gTOP3 | zh_TW |
| dc.description.abstract | Giardiasis is a common disease caused by Giardia lamblia. The symptoms of giardiasis included diarrhoea, abdominal pain, weight loss, and malabsorption. Giardia has two major types: trophzoite and cyst. In trophzoite form, it’s can parasitize in the host’s instestinal epithelium and cause giardiasis. Cysts form of G. lamblia are passed in the faeces and protect Giardia against the environment. Both trophozoite with two nuclei or cyst with four tetraploid nuclei all need to proceed accuratent mechanism for DNA. When DNA is carrying out replication, transcription, recombination, chromosome compaction and segregation, some topological problems would occur these processes would result some topological problems. Therefore, topoisomerases can be applied to solve these construction problems to ensure DNA is normal.
We have analyzed the G. lamblia genome and found topoisomerase Ⅲalpha(gTOP3 | en |
| dc.description.provenance | Made available in DSpace on 2021-06-08T01:13:49Z (GMT). No. of bitstreams: 1 ntu-103-R01445206-1.pdf: 3296665 bytes, checksum: 2bf9dd9c560fd07a233d0988f2eaea8b (MD5) Previous issue date: 2014 | en |
| dc.description.tableofcontents | 口試委員審定書 i
誌謝 ii 中文摘要 iii abstract iv 目錄 vi 第一章 前言 1 1.1 梨形鞭毛蟲簡介 1 1.2 DNA topoisomerase 2 1.3 Topoisomerase Type IA 3 1.4 研究動機 5 第二章 材料與方法 6 2.1 梨形鞭毛蟲的細胞株培養(G. lamblia Cuture) 6 2.2 轉殖質體的建構(Plasmid Construction) 6 2.2.1 5’∆5N-pac 6 2.2.2 pPgTOP3a 6 2.2.3 pPgTOP3a m1 7 2.2.4 pPgTOP3a m2 8 2.2.5 pPgTOP3a m3 8 2.3 轉殖質體的轉型與萃取 9 2.3.1 質體的轉型(transformation) 9 2.3.2 質體的萃取(plasmid preparation) 9 2.4 梨形鞭毛蟲的轉染與選殖(Transfection and Selection) 9 2.5 重組蛋白質gTOP3a及突變gTOP3a表現載體建構 10 2.5.1 重組gTOP3a蛋白質的表現載體建構 10 2.5.2 重組gTOP3a m1蛋白質的表現載體建構 10 2.5.3 重組gTOP3a m2蛋白質的表現載體建構 11 2.5.4 重組gTOP3a m3蛋白質的表現載體建構 11 2.6 重組蛋白質的表現與純化 11 2.7 反轉錄聚合酶鍊式反應(Semi-quantitative RT-PCR Analysis，RT-PCR) 12 2.8 及時定量反轉錄聚合酶鍊式反應(Quantitative real-time polymerase chain reaction，Q-PCR) 12 2.9 西方點墨法(Western Blot Analysis)與Coomassie Blue染色 13 2.9.1 TCA蛋白質沉澱(TCA protein precipitation) 13 2.9.2 西方點墨法(Western Blot Analysis) 14 2.9.3 Coomassie Blue染色 14 2.10免疫螢光染色(Immunofluorescence Assay) 15 2.11 切割DNA活性分析(DNA Cleavage Assays) 15 2.11.1負超螺旋DNA切割活性分析 15 2.12囊體計數(Cyst Count) 15 第三章 實驗結果 17 3.1 分析梨形鞭毛蟲topoisomeraseⅢa(gTOP3a)胺基酸序列 17 3.2 梨形鞭毛蟲之gTOP3a基因在不同時期的表現量 18 3.3 梨形鞭毛蟲之gTOP3a對於DNA的切割能力 19 3.4 梨形鞭毛蟲之gTOP3a蛋白質可抑制囊體壁蛋白cwp1的表現量。 19 3.5 梨形鞭毛蟲之gTOP3a蛋白質對於其他基因的RNA表現量之影響。 19 3.6 建立gTOP3a的突變細胞來分析gTOP3a的活性區域。 20 第四章 討論 22 4.1 gTOP3a在不同時期表現量 22 4.2 gTOP3a的活性區分析 22 4.3 gTOP3a影響囊體壁蛋白cwp1的表現。 23 4.4 gTOP3a影響VSP的表現量。 24 4.5 gTOP3a對於Casein kinases基因之表現量 24 4.6 梨形鞭毛蟲gTOP3a基因啟動子區域不具有調控囊體化相關基因的轉錄因子之結合位。 25 4.7 gTOP3a與其他蛋白之交互作用 25 附圖 26 附表 45 參考資料 50 | |
| dc.language.iso | zh-TW | |
| dc.title | 梨形鞭毛蟲拓樸異構酶Ⅲalpha對於囊體壁基因的表現之影響 | zh_TW |
| dc.title | The effect of topoisomerase Ⅲalpha on the cyst wall gene expression of Giardia lamblia | en |
| dc.type | Thesis | |
| dc.date.schoolyear | 102-2 | |
| dc.description.degree | 碩士 | |
| dc.contributor.oralexamcommittee | 李財坤(Tsai-Kun Li),詹迺立(Nei-Li, Chan) | |
| dc.subject.keyword | 梨形鞭毛蟲,囊體化,拓樸異構?,cyst wall protein(cwp),myb2, | zh_TW |
| dc.subject.keyword | Giardia lamblia,Encystation,topoisomerases,cyst wall protein(cwp),myb2, | en |
| dc.relation.page | 53 | |
| dc.rights.note | 未授權 | |
| dc.date.accepted | 2014-08-14 | |
| dc.contributor.author-college | 醫學院 | zh_TW |
| dc.contributor.author-dept | 微生物學研究所 | zh_TW |
| 顯示於系所單位： | 微生物學科所 | |
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