請用此 Handle URI 來引用此文件：
http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/81095完整後設資料紀錄
| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 廖淑貞(Shwu-Jen Liaw) | |
| dc.contributor.author | Lu Wang | en |
| dc.contributor.author | 王璐 | zh_TW |
| dc.date.accessioned | 2022-11-24T03:30:18Z | - |
| dc.date.available | 2021-08-31 | |
| dc.date.available | 2022-11-24T03:30:18Z | - |
| dc.date.copyright | 2021-08-31 | |
| dc.date.issued | 2021 | |
| dc.date.submitted | 2021-08-20 | |
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| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/81095 | - |
| dc.description.abstract | 奇異變形桿菌(P. mirabilis)為人體一重要的尿道致病菌，其常引發尿導管相關的尿道感染。當尿道感染發生時人體為抵禦病菌的入侵會提升尿液中的銅濃度，其中P. mirabilis所引發的UTI可使尿液中銅濃度上升至約0.5 μM，同時清除外來致病菌的巨噬細胞中銅濃度會提升至約500 μM來加强對致病菌的毒殺作用。過量的銅會對菌體造成傷害，故細菌會啓動抗銅機制來維持菌體内銅濃度之恆定。目前對P. mirabilis的抗銅機制所知甚少，故本篇欲探究P. mirabilis的抗銅機制。 首先利用跳躍子突變法從約15000株突變株中篩選得到5株對銅感受性提高之突變株，經過基因分析後得到cueR突變株及copA突變株；同時以blast方式來尋找P. mirabilis中存在的抗銅基因，並結合實驗室之前transcriptome之結果推測出Cue系統於P. mirabilis的抗銅機制中扮演著重要作用。於是利用reporter assay及qPCR確認了P. mirabilis之CueR可正向調控抗銅相關基因cueO、 copA及copG之表達，而AAS之結果亦印證CueR可幫助菌體維持菌體内之銅濃度恆定。 接著探究CueR在0.5 μM及500 μM這兩個已知的生理銅濃度下對菌體毒力因子之影響，本篇結果表明無銅或0.5 μM銅離子環境并不影響cueR突變株生長狀況及細胞外毒力因子之表達，但於500 μM 銅離子環境下，cueR突變株相較於野生株之生長狀況、尿素酶活性、生物膜形成能力、表面移行能力及泳動能力皆顯著下降，由reporter及qPCR之分析可知cueR突變株因鞭毛形成相關基因flhDC的轉錄及fimbriae中與感染力相關之基因mrpA、pmfA、fim14、pmpA之表現量顯著下降，進而造成其運動性及細胞黏附能力下降。而在預處理銅後其抗氧化能力及巨噬細胞内存活率顯著上升可能與scsA表現量增加有關，但在未預先處理銅時cueR突變株之抗氧化能力及巨噬細胞内存活率則顯著低於野生株，同時小鼠實驗亦證實cueR突變株於小鼠膀胱及腎臟中的定植能力均顯著下降。 綜上可知於銅環境下CueR不僅可正向調節cueO、copA及copG之基因表達來維持菌體内銅濃度水平，亦可影響尿素酶活性、運動力、生物膜形成能力、上皮細胞黏附能力、巨噬細胞内存活能力及其定植能力等多種毒力因子。因此CueR在P. mirabilis之抵抗銅壓力及維持毒力因子中均扮演著重要角色。 | zh_TW |
| dc.description.provenance | Made available in DSpace on 2022-11-24T03:30:18Z (GMT). No. of bitstreams: 1 U0001-1808202122062400.pdf: 2906024 bytes, checksum: 027b49ae30ba0987a3dee84888c1c165 (MD5) Previous issue date: 2021 | en |
| dc.description.tableofcontents | 致謝 i 摘要 ii 英文摘要 iv 目錄 vi 第一章緒論 1 第一節奇異變形桿菌(Proteus mirabilis)的基本介紹 1 第二節P. mirabilis 的毒力因子 1 第三節P. mirabilis 的表面移行能力（swarming） 4 第四節銅之生物特性 4 第五節銅之生理意義 6 第六節菌體内之銅恆定系統 7 第七節CueR 之基本特性 8 第八節研究動機與目的 9 第二章實驗設計、材料與方法 10 第一節實驗設計 10 第二節實驗材料 11 第三節構建突變株、互補株及過度表達株 13 第四節突變基因之鑑定 21 第五節表現型（phenotype）及毒力因子（virulence factors）分析 27 第六節基因表達 43 第三章實驗結果 48 第一節篩選對銅高感受性之突變株 48 第二節P. mirabilis 之銅恆定系統 51 第三節cueR 突變株之表現型分析 56 第四節討論與結論 71 參考資料 76 第四章表 82 附錄 86 | |
| dc.language.iso | zh-TW | |
| dc.subject | 奇異變形桿菌 | zh_TW |
| dc.subject | 銅 | zh_TW |
| dc.subject | 銅調控子CueR | zh_TW |
| dc.subject | 毒力因子 | zh_TW |
| dc.subject | 尿道致病菌 | zh_TW |
| dc.subject | Proteus mirabilis | en |
| dc.subject | virulence factor | en |
| dc.subject | copper regulator CueR | en |
| dc.subject | copper | en |
| dc.subject | uropathogen | en |
| dc.title | 探討尿道致病性奇異變形桿菌之抗銅機制-銅結合蛋白質CueR抗銅及調控毒力因子之研究 | zh_TW |
| dc.title | Characterization of copper resistance in uropathogenic Proteus mirabilis-the role of copper regulator CueR in copper resistance and virulence | en |
| dc.date.schoolyear | 109-2 | |
| dc.description.degree | 碩士 | |
| dc.contributor.oralexamcommittee | 賴信志(Hsin-Tsai Liu),邱浩傑(Chih-Yang Tseng) | |
| dc.subject.keyword | 尿道致病菌,奇異變形桿菌,銅,銅調控子CueR,毒力因子, | zh_TW |
| dc.subject.keyword | uropathogen,Proteus mirabilis,copper,copper regulator CueR,virulence factor, | en |
| dc.relation.page | 98 | |
| dc.identifier.doi | 10.6342/NTU202102493 | |
| dc.rights.note | 同意授權(限校園內公開) | |
| dc.date.accepted | 2021-08-20 | |
| dc.contributor.author-college | 醫學院 | zh_TW |
| dc.contributor.author-dept | 醫學檢驗暨生物技術學研究所 | zh_TW |
| 顯示於系所單位： | 醫學檢驗暨生物技術學系 | |
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