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http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/30567完整後設資料紀錄
| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 王錦堂 | |
| dc.contributor.author | Han-Chi Fang | en |
| dc.contributor.author | 方涵琪 | zh_TW |
| dc.date.accessioned | 2021-06-13T02:09:01Z | - |
| dc.date.available | 2012-07-12 | |
| dc.date.copyright | 2007-07-12 | |
| dc.date.issued | 2007 | |
| dc.date.submitted | 2007-06-30 | |
| 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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Community-acquired Klebsiella pneumoniae bacteremia: global differences in clinical patterns. Emerg Infect Dis 8, 160-166. Lai, Y.C., Peng, H.L., and Chang, H.Y. (2003). RmpA2, an activator of capsule biosynthesis in Klebsiella pneumoniae CG43, regulates K2 cps gene expression at the transcriptional level. J Bacteriol 185, 788-800. Link, A.J., Phillips, D., and Church, G.M. (1997). Methods for generating precise deletions and insertions in the genome of wild-type Escherichia coli: application to open reading frame characterization. J Bacteriol 179, 6228-6237. Liu, Y.C., Cheng, D.L., and Lin, C.L. (1986). Klebsiella pneumoniae liver abscess associated with septic endophthalmitis. Arch Intern Med 146, 1913-1916. Ma, L.C., Fang, C.T., Lee, C.Z., Shun, C.T., and Wang, J.T. (2005). Genomic heterogeneity in Klebsiella pneumoniae strains is associated with primary pyogenic liver abscess and metastatic infection. J Infect Dis 192, 117-128. 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Tsai, W.C., Strieter, R.M., Zisman, D.A., Wilkowski, J.M., Bucknell, K.A., Chen, G.H., and Standiford, T.J. (1997). Nitric oxide is required for effective innate immunity against Klebsiella pneumoniae. Infect Immun 65, 1870-1875. Wacharotayankun, R., Arakawa, Y., Ohta, M., Tanaka, K., Akashi, T., Mori, M., and Kato, N. (1993). Enhancement of extracapsular polysaccharide synthesis in Klebsiella pneumoniae by RmpA2, which shows homology to NtrC and FixJ. Infect Immun 61, 3164-3174. Wang, J.H., Liu, Y.C., Lee, S.S., Yen, M.Y., Chen, Y.S., Wang, J.H., Wann, S.R., and Lin, H.H. (1998). Primary liver abscess due to Klebsiella pneumoniae in Taiwan. Clin Infect Dis 26, 1434-1438. Yeh, K.M., Kurup, A., Siu, L.K., Koh, Y.L., Fung, C.P., Lin, J.C., Chen, T.L., Chang, F.Y., and Koh, T.H. (2007). Capsular serotype K1 or K2, rather than magA and rmpA, is a major virulence determinant for Klebsiella pneumoniae liver abscess in Singapore and Taiwan. J Clin Microbiol 45, 466-471. Yu, W.L., Ko, W.C., Cheng, K.C., Lee, H.C., Ke, D.S., Lee, C.C., Fung, C.P., and Chuang, Y.C. (2006). Association between rmpA and magA genes and clinical syndromes caused by Klebsiella pneumoniae in Taiwan. Clin Infect Dis 42, 1351-1358. 方啟泰 (2002) 造成肝膿瘍之侵襲性克雷伯氏肺炎桿菌菌株致病基因的研究. 臨床醫學研究所. 博士論文 台北: 國立台灣大學醫學院, pp. 95. | |
| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/30567 | - |
| dc.description.abstract | 近二十年來，在台灣出現了一種新型的侵襲性克雷伯氏肺炎桿菌，會引起典型的臨床表現－原發性肝膿瘍 (primary liver abscess, PLA) 合併菌血症，已知克雷伯氏肺炎桿菌的致病因子之中包含莢膜與黏性型態，莢膜能夠保護細菌不受吞噬細胞 (phagocytes) 的吞噬作用及血清中補體 (complement) 的殺菌作用所摧毀。
本研究發現rmpA基因的分布在PLA菌株較non-PLA菌株為高 (100% vs. 22%)。rmpA-突變株已失去了野生株的高黏性特徵，且相較於野生株降低對小鼠的致死能力，而rmpA異位互補株 (trans-complementation) 則使得黏性回復。利用克雷伯氏肺炎桿菌基因微陣列，分析野生株與rmpA-突變株基因表現之差異來尋找RmpA直接或間接調控的致病因子，經過RT-qPCR的確認，找到在rmpA-突變株表現量下降，並且在rmpA異位互補株表現量上升的基因：莢膜生合成基因組 (cps gene cluster) 與nirB、nirD (nitrite reductase) 基因。已知莢膜生合成基因組為重要致病因子，且會受到RmpA之相似蛋白質RmpA2的活化。而nirB、nirD基因的表現並不受莢膜合成的影響，可能為RmpA影響莢膜以外的致病因子。然而小鼠實驗顯示nirBD-突變株對小鼠的致死能力與野生株無差異。因此只有莢膜生合成基因組為RmpA所調控的致病因子。 | zh_TW |
| dc.description.abstract | In the past 20 years, a new type of invasive Klebsiella pneumoniae disease has emerged in Taiwan that typically presents a community-acquired primary liver abscess (PLA) with sepsis and bacteremia. It has been known that two of the virulent factors of K. pneumoniae are capsule and mucoid phenotype. The capsular polysaccharide protect against the bactericidal effect of serum complement and against ingestion and killing by professional phagocytes.
In this study, we found rmpA was significantly more prevalent in PLA strains. Furthermore, the rmpA- mutants lost the hypermucoviscosity and resulted in an increase of LD50 compared to wild-type strains in a mouse intraperitoneal infection model. The mucoviscosity was restored by complementation using a rmpA-containing plasmid. The above data suggested that rmpA was correlated to virulence resulted from K. pneumoniae. In order to find RmpA-regulated virulent factors, we compared RNA expression profiles between wild-type strains and rmpA- mutants by using microarray analysis. The microarray data were confirmed by RT-qPCR, and we found cps (capsular polysaccharide) gene cluster and nir operon (nitrite reductase) expression were decreased in rmpA- mutants but increased in rmpA-complementation strains. It has been well documented that CPS is an important virulent factor activated by RmpA homolog, RmpA2. The expression of nir operon is not correlated to CPS but regulated by RmpA. The nir operon may be involved in CPS-independent virulence mechanism regulated by RmpA. However, there was no significant difference in LD50 between wild-type strains and nirBD- mutants. Therefore, only cps gene clusters are found to be virulent genes regulated by RmpA. | en |
| dc.description.provenance | Made available in DSpace on 2021-06-13T02:09:01Z (GMT). No. of bitstreams: 1 ntu-96-R94445102-1.pdf: 813209 bytes, checksum: 0d0972dcbadbc4de80ac990f992eeaf6 (MD5) Previous issue date: 2007 | en |
| dc.description.tableofcontents | 口試委員會審定書……………………………..…………...………………………Ⅰ
致謝……………………………..……………………………………...……………Ⅱ 中文摘要……………………………..………………………………………...……Ⅲ 英文摘要……………………………………………………………………...……..Ⅳ 目錄……………………………..………………………………………………...…Ⅵ 圖目錄……………………………..………………………………...………………Ⅷ 表目錄……………………………..………………...………………………………Ⅹ 第一章、 緒論……………………………..………………..………………………1 第二章、 材料與方法………….………………………………...…………………5 1. 克雷伯氏肺炎桿菌菌株………………………………………….…...……5 2. 聚合酶鏈鎖反應 (polymerase chain reaction, PCR)……………….…….…5 3. 萃取克雷伯氏肺炎桿菌基因體DNA (genomic DNA)…………….…...…6 4. 萃取克雷伯氏肺炎桿菌total RNA……………………………..……...…..6 5. 克雷伯氏肺炎桿菌基因微陣列………………………………………....…8 6. 反轉錄定量聚合酶鏈鎖反應(RT-qPCR)………………………...…….…11 7. rmpA異位互補 (trans-complementation)………………….……………..12 8. 拉絲測試 (String test)……………………………………....…………….12 9. 血清抗性試驗 (Serum resistance assay)………………………………....12 10. 建構克雷伯氏肺炎桿菌ΔnirBD突變株…………………………...……13 11. 動物實驗…………………………………………………...….…………15 第三章、 結果……………………………………………………………...……...17 1. 觀察rmpA在臨床菌株中的分佈比例………………...…………….……17 2. 分析克雷伯氏肺炎桿菌NTUH-K2044菌株基因體上之rmpA/rmpA2序列……………………………….....................................................................17 3. 比較NTUH-K2044菌株基因體上兩套rmpA基因的RNA表現量……………………………….....................................................................18 4. 建構NTUH-K3-25異位互補株 (trans-complementation) 並觀察其表型……………………………….....................................................................19 5. 探討NTUH-K2044野生株及NTUH-K3-25突變株對於小鼠的致病力……………………………….....................................................................19 6. 利用克雷伯氏肺炎桿菌基因微陣列尋找rmpA所調控的基因……………………………….....................................................................20 7. 利用RT-qPCR確認微陣列結果，並從中尋找出rmpA可能調控的基因……………………………….....................................................................20 8. 建構ΔnirBD突變株，並探討NTUH-K2044野生株及ΔnirBD突變株對於小鼠的致病力…………………………………………………….……….22 第四章、 總結與討論……………………………………………………………..23 第五章、 參考文獻………………………………………………...….…………..61 | |
| dc.language.iso | zh-TW | |
| dc.subject | 克雷伯氏肺炎桿菌 | zh_TW |
| dc.subject | 莢膜 | zh_TW |
| dc.subject | K1 | zh_TW |
| dc.subject | rmpA | zh_TW |
| dc.subject | nir operon | zh_TW |
| dc.subject | nir operon | en |
| dc.subject | Klebsiella pneumoniae | en |
| dc.subject | capsule | en |
| dc.subject | K1 | en |
| dc.subject | rmpA | en |
| dc.title | 研究rmpA 在克雷伯氏肺炎桿菌中的功能 | zh_TW |
| dc.title | Identification of the role of rmpA in Klebsiella pneumoniae | en |
| dc.type | Thesis | |
| dc.date.schoolyear | 95-2 | |
| dc.description.degree | 碩士 | |
| dc.contributor.oralexamcommittee | 鄧述諄,賴信志 | |
| dc.subject.keyword | 克雷伯氏肺炎桿菌,莢膜,K1,rmpA,nir operon, | zh_TW |
| dc.subject.keyword | Klebsiella pneumoniae,capsule,K1,rmpA,nir operon, | en |
| dc.relation.page | 68 | |
| dc.rights.note | 有償授權 | |
| dc.date.accepted | 2007-07-02 | |
| 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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