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http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/38049完整後設資料紀錄
| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 王錦堂 | |
| dc.contributor.author | Pei-Fang Hsieh | en |
| dc.contributor.author | 謝佩芳 | zh_TW |
| dc.date.accessioned | 2021-06-13T15:58:45Z | - |
| dc.date.available | 2011-08-13 | |
| dc.date.copyright | 2008-08-13 | |
| dc.date.issued | 2008 | |
| dc.date.submitted | 2008-05-19 | |
| dc.identifier.citation | Aguilar, J., Cruz, A., and Ortega, C. (1994) [Multiple hepatic and pulmonary abscesses caused by Klebsiella pneumoniae]. Enferm Infecc Microbiol Clin 12: 270-271.
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| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/38049 | - |
| dc.description.abstract | 本論文利用基因微陣列方法來研究克雷伯氏肺炎桿菌於健康人血清（human serum, HS）中之血清誘發基因的表現，發現到三組受血清誘發表現的鐵離子攝取系統與臨床上造成社區型細菌性肝膿瘍之克雷伯氏肺炎桿菌具有關聯性：Yersinia high-pathogenicity island（Yersinia HPI）、iucABCDiutA 和 iroA 基因群組，利用交互飼養試驗（cross-feeding assay），已證實這 3 段基因群組之鐵離子攝取系統的功能。只有注射 irp2 iuc iroA triple mutant 此多重基因剔除菌株於鼷鼠動物模式會降低此菌的致病力。由克雷伯氏肺炎桿菌 NTUH-K2044 菌株之全基因體序列分析，共發現 10 組鐵離子攝取系統，其中有 7 組皆為 TonB-依賴型之鐵離子攝取系統，包括 Yersinia HPI 、 iucABCDiutA 和iroA 基因群組。於鼷鼠動物模式中， tonB 基因剔除菌株大幅度地減弱此菌的致病力。以腹膜腔注射 tonB 基因剔除菌株來免疫鼷鼠，結果顯示鼷鼠血清中之莢膜抗體陽轉且對於後續原生菌株之感染具保護力。總結，鐵離子攝取系統是造成社區型細菌性肝膿瘍之克雷伯氏肺炎桿菌主要的血清誘發基因，雖然在 NTUH-K2044 菌株中存在很多組的鐵離子攝取系統，只有 irp2 iuc iroA 之多重基因剔除菌株才能減弱此菌的致病力，活性減毒 tonB 基因剔除菌株具有作為疫苗株之潛力，可誘發鼷鼠產生具顯著性之對抗原生菌株的保護性免疫反應。 | zh_TW |
| dc.description.abstract | Klebsiella pneumoniae was stimulated by human serum and gene expression was analyzed by microarray. Three putative iron acquisition systems, Yersinia high-pathogenicity island（Yersinia HPI）, iucABCDiutA and iroA(iroNDCB), that increased in expression and predominate in PLA-associated K. pneumoniae strains were identified. By siderophore uptake assays, these three systems were confirmed to be siderophore-dependent iron acquisition systems. Only the irp2 iuc iroA triple mutant showed decreased virulence in mice. Full genome analysis of K. pneumoniae NTUH-K2044 identified 10 putative iron uptake systems. Seven of them were TonB-dependent, including Yersinia HPI, iucABCDiutA and iroA. A tonB deletion mutant was demonstrated profound attenuation of virulence. Immunization with the tonB mutant showed sero-conversion of extracellular polysaccharides antibody and protective efficacy against subsequent exposure to the parental strain. Taken together, iron uptake systems were the highly up-regulated genes in K. pneumoniae in response to sera. Although there are multiple iron transporter systems in NTUH-K2044, a mutation of all three loci, irp2, iuc, and iroA, is necessary to decrease virulence. The tonB mutant is a potential vaccine candidate because it can induce significant protective immune response against wild-type strain challenge. | en |
| dc.description.provenance | Made available in DSpace on 2021-06-13T15:58:45Z (GMT). No. of bitstreams: 1 ntu-97-D93445004-1.pdf: 753923 bytes, checksum: a7fd29e5289b20178af750ff8d472b0b (MD5) Previous issue date: 2008 | en |
| dc.description.tableofcontents | 目錄
口試委員會審定書…………I 誌謝…………II 中文摘要…………III 英文摘要…………IV 第一章 緒論…………1 1.1 克雷伯氏肺炎桿菌與社區型細菌性肝膿瘍…………1 1.2 致病因子…………2 1.3 鐵離子來源…………2 1.4 鐵結合因子、原血紅素結合因子和 TonB 蛋白質…………3 1.5 TonB-依賴型之鐵離子攝取系統：yersiniabactin（Ybt）、aerobactin（iucABCDiutA）和 salmochelin（iroBCDN）…………5 1.6 研究目的…………6 第二章 材料及方法…………7 2.1 材料…………7 2.1.1 克雷伯氏肺炎桿菌菌株…………7 2.1.2 基因微陣列…………8 2.1.3 血清…………8 2.1.4 大腸桿菌及耶氏桿菌菌株…………8 2.1.5 培養基…………9 2.1.6 抗生素…………10 2.1.7 引子…………10 2.2 方法…………10 2.2.1 克雷伯氏肺炎桿菌 total RNA 的抽取…………10 2.2.2 基因微陣列探針的合成…………11 2.2.3 基因微陣列的雜交…………12 2.2.4 基因微陣列呈色反應及影像分析…………12 2.2.5 聚合酶鏈鎖反應…………13 2.2.6 基因剔除菌株的建構…………13 2.2.7 異位補回…………14 2.2.8 交互飼養試驗…………14 2.2.9 生長測試…………15 2.2.10 拉絲測試…………15 2.2.11 血清抗性試驗…………15 2.2.12 莢膜血清型分型…………16 2.2.13 動物實驗…………16 2.2.14 酵素連結免疫吸附分析…………18 2.2.15 免疫轉漬法…………18 第三章 結果…………19 3.1 基因微陣列分析結果…………19 3.2 Yersinia HPI、iucABCDiutA 和 iroA基因群組在臨床克雷伯氏肺炎桿菌菌株的分佈情形…………19 3.3 Yersinia HPI、iucABCDiutA、iroA(iroNDCB)、kfu/PTS、magA和 allS等六個基因片段與造成社區型細菌性肝膿瘍菌株間的關聯…………20 3.4 irp2、iuc或是iroA單一基因剔除菌株、irp2 iuc iroA triple mutant、irp2 iuc iro kfu quadruple mutant等多重基因剔除菌株和其相對應之基因補回菌株之特性分析…………20 3.5 克雷伯氏肺炎桿菌NTUH-K2044之鐵離子攝取系統…………21 3.6 TonB蛋白質之功能分析…………22 3.7 基因剔除菌株之免疫途徑與效力…………22 第四章 總結與討論…………24 4.1 血清誘發基因…………24 4.2 鐵離子攝取系統對細菌致病力的影響…………25 4.3 糖尿病與社區型細菌性肝膿瘍之關連性…………27 4.4 社區型細菌性肝膿瘍之可能感染模式…………28 4.5 tonB 基因剔除菌株作為活性減毒株疫苗之免疫保護力…………30 表目錄 表一 本研究中所使用之菌株和質體…………32 表二 本研究中所使用之引子…………33 表三 克雷伯氏肺炎桿菌 NTUH-K2044 之血清誘發基因…………35 表四 Yersinia HPI、iucABCDiutA、iroA(iroNDCB)、kfu/PTS、magA 和allS 等六大基因片段在臨床克雷伯氏肺炎桿菌菌株間的分佈情形…………36 表五 克雷伯氏肺炎桿菌 NTUH-K2044 鐵離子攝取系統相關基因剔除株之特性分析…………37 表六 克雷伯氏肺炎桿菌 NTUH-K2044 之 IroN 蛋白質序列相似度及對於 catecholate 類鐵結合因子之利用…………38 表七 克雷伯氏肺炎桿菌 NTUH-K2044 及其基因剔除株於鼷鼠動物模式之 LD50劑量…………39 表八 克雷伯氏肺炎桿菌 NTUH-K2044 之鐵離子攝取系統…………40 表九 以腹膜腔注射克雷伯氏肺炎桿菌 NTUH-K2044 基因剔除株免疫鼷鼠之後再感染原生菌株…………41 表十 以胃內感染克雷伯氏肺炎桿菌 NTUH-K2044 基因剔除株免疫鼷鼠之後再感染原生菌株…………42 圖目錄 圖一 血清誘發克雷伯氏肺炎桿菌基因體表現之基因微陣列流程圖…………43 圖二 血清誘發克雷伯氏肺炎桿菌NTUH-K2044基因微陣列分析結果…………44 圖三 反轉錄-定量 PCR…………45 圖四 30-kb Yersinia HPI、8-kb iucABCDiutA、9-kb iroA(iroBCDN)、iroA(iroNDCB) 和6-kb hmuRSTUV 基因片段之圖示…………46 圖五 NTUH-K2044 之鐵離子攝取系統相關基因剔除株之建構圖示…………47 圖六 NTUH-K2044及其基因剔除株於含有鐵螯合因子2,2’-dipyridyl (DIP) 之 nutrient broth (NB) 中的生長情形…………48 圖七 交互飼養試驗…………49 圖八 以胃內感染方式餵食鼷鼠 1 ´106 CFU 克雷伯氏肺炎桿菌 NTUH-K2044 及其基因剔除株之存活情形…………50 圖九 利用對向免疫擴散法觀察NTUH-K2044 各鐵離子攝取系統相關基因剔除株之莢膜血清型 K1 的表現情形…………51 圖十 tonB 基因剔除株之血清抗性分析…………52 圖十一比較NTUH-K2044 原生菌株或是 tonB 基因剔除株於鼷鼠動物體內的生長情形…………53 圖十二以腹膜腔注射 tonB 基因剔除株進行免疫之後再感染次致死劑量之 NTUH-K2044 原生菌株鼷鼠的存活情形…………54 圖十三tonB 基因剔除菌株免疫前後鼷鼠血清中 anti-EPS IgG 抗體陽轉情形…………55 第五章 參考文獻…………56 | |
| dc.language.iso | zh-TW | |
| dc.title | 克雷伯氏肺炎桿菌之血清誘發鐵離子攝取系統和 TonB 蛋白質對其致病力的影響 | zh_TW |
| dc.title | Serum-induced iron-acquisition systems and TonB contribute to virulence in Klebsiella pneumoniae causing primary pyogenic liver abscess | en |
| dc.type | Thesis | |
| dc.date.schoolyear | 96-2 | |
| dc.description.degree | 博士 | |
| dc.contributor.oralexamcommittee | 伍安怡,鄧麗珍,賴信志,顧家綺 | |
| dc.subject.keyword | 克雷伯氏肺炎桿菌,社區型細菌性肝膿瘍,鐵離子攝取系統,鐵結合因子,tonB 基因, | zh_TW |
| dc.subject.keyword | Klebsiella pneumoniae,community-acquired pyogenic liver abscess(PLA),iron acquisition systems,siderophores,tonB, | en |
| dc.relation.page | 68 | |
| dc.rights.note | 有償授權 | |
| dc.date.accepted | 2008-05-21 | |
| 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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