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完整後設資料紀錄
DC 欄位 | 值 | 語言 |
---|---|---|
dc.contributor.advisor | 王錦堂(Jin-Town Wang) | |
dc.contributor.author | You-Ci Chen | en |
dc.contributor.author | 陳又綺 | zh_TW |
dc.date.accessioned | 2021-06-15T04:17:02Z | - |
dc.date.available | 2015-03-12 | |
dc.date.copyright | 2010-03-12 | |
dc.date.issued | 2009 | |
dc.date.submitted | 2009-12-27 | |
dc.identifier.citation | Aguilar, J., Cruz, A., and Ortega, C. (1994). [Multiple hepatic and pulmonary abscesses
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Fang, C.T., Chuang, Y.P., Shun, C.T., Chang, S.C., and Wang, J.T. (2004). A novel virulence gene in Klebsiella pneumoniae strains causing primary liver abscess and septic metastatic complications. J Exp Med 199, 697-705. Fang, C.T., Lai, S.Y., Yi, W.C., Hsueh, P.R., Liu, K.L., and Chang, S.C. (2007). Klebsiella pneumoniae genotype K1: an emerging pathogen that causes septic ocular or central nervous system complications from pyogenic liver abscess. Clin Infect Dis 45, 284-293. Fang, H.C. (2007). Identification of the role of rmpA in Klebsiella pneumoniae. In 微生 物學研究所 碩士論文 (台北國立台灣大學醫學院). Fung, C.P., Chang, F.Y., Lee, S.C., Hu, B.S., Kuo, B.I., Liu, C.Y., Ho, M., and Siu, L.K. (2002). A global emerging disease of Klebsiella pneumoniae liver abscess: is serotype K1 an important factor for complicated endophthalmitis? Gut 50, 420-424. Karama, E.M., Willermain, F., Janssens, X., Claus, M., Van den Wijngaert, S., Wang, J.T., Verougstraete, C., and Caspers, L. (2008). Endogenous endophthalmitis pyogenic liver abscess. J Infect Dis 193, 645-654. Clements, A., Gaboriaud, F., Duval, J.F., Farn, J.L., Jenney, A.W., Lithgow, T., Wijburg, O.L., Hartland, E.L., and Strugnell, R.A. (2008). The major surface-associated saccharides of Klebsiella pneumoniae contribute to host cell association. PLoS One 3, e3817. Cortes, G., Borrell, N., de Astorza, B., Gomez, C., Sauleda, J., and Alberti, S. (2002). Molecular analysis of the contribution of the capsular polysaccharide and the lipopolysaccharide O side chain to the virulence of Klebsiella pneumoniae in a murine model of pneumonia. Infect Immun 70, 2583-2590. Couez, D., Libon, E., Wasteels, M., Derue, G., and Gilbeau, J.P. (1991). Klebsiella pneumoniae liver abscess with septic endophthalmitis. The role of computed tomography. J Belge Radiol 74, 41-44. Cryz, S.J., Jr., Furer, E., and Germanier, R. (1984). Protection against fatal Klebsiella pneumoniae burn wound sepsis by passive transfer of anticapsular polysaccharide. Infect Immun 45, 139-142. Domenico, P., Diedrich, D.L., and Straus, D.C. (1985). Extracellular polysaccharide production by Klebsiella pneumoniae and its relationship to virulence. Can J Microbiol 31, 472-478. Domenico, P., Tomas, J.M., Merino, S., Rubires, X., and Cunha, B.A. (1999). Surface antigen exposure by bismuth dimercaprol suppression of Klebsiella pneumoniae capsular polysaccharide. Infect Immun 67, 664-669. Erbing, C., Kenne, L., Lonngren, J., and Sutherland, I.W. (1976). Structural studies of the capsular polysaccharide from Klebsiella type 1. Carbohydr Res 50, 115-120. Fang, C.T., Chuang, Y.P., Shun, C.T., Chang, S.C., and Wang, J.T. (2004). A novel virulence gene in Klebsiella pneumoniae strains causing primary liver abscess and septic metastatic complications. J Exp Med 199, 697-705. Fang, C.T., Lai, S.Y., Yi, W.C., Hsueh, P.R., Liu, K.L., and Chang, S.C. (2007). Klebsiella pneumoniae genotype K1: an emerging pathogen that causes septic ocular or central nervous system complications from pyogenic liver abscess. Clin Infect Dis 45, 284-293. Fang, H.C. (2007). Identification of the role of rmpA in Klebsiella pneumoniae. In 微生 物學研究所 碩士論文 (台北國立台灣大學醫學院). Fung, C.P., Chang, F.Y., Lee, S.C., Hu, B.S., Kuo, B.I., Liu, C.Y., Ho, M., and Siu, L.K. (2002). A global emerging disease of Klebsiella pneumoniae liver abscess: is serotype K1 an important factor for complicated endophthalmitis? Gut 50, 420-424. Karama, E.M., Willermain, F., Janssens, X., Claus, M., Van den Wijngaert, S., Wang, J.T., Verougstraete, C., and Caspers, L. (2008). Endogenous endophthalmitis pyogenic liver abscess. J Infect Dis 193, 645-654. Clements, A., Gaboriaud, F., Duval, J.F., Farn, J.L., Jenney, A.W., Lithgow, T., Wijburg, O.L., Hartland, E.L., and Strugnell, R.A. (2008). The major surface-associated saccharides of Klebsiella pneumoniae contribute to host cell association. PLoS One 3, e3817. Cortes, G., Borrell, N., de Astorza, B., Gomez, C., Sauleda, J., and Alberti, S. (2002). Molecular analysis of the contribution of the capsular polysaccharide and the lipopolysaccharide O side chain to the virulence of Klebsiella pneumoniae in a murine model of pneumonia. Infect Immun 70, 2583-2590. Couez, D., Libon, E., Wasteels, M., Derue, G., and Gilbeau, J.P. (1991). Klebsiella pneumoniae liver abscess with septic endophthalmitis. The role of computed tomography. J Belge Radiol 74, 41-44. Cryz, S.J., Jr., Furer, E., and Germanier, R. (1984). Protection against fatal Klebsiella pneumoniae burn wound sepsis by passive transfer of anticapsular polysaccharide. Infect Immun 45, 139-142. Domenico, P., Diedrich, D.L., and Straus, D.C. (1985). Extracellular polysaccharide production by Klebsiella pneumoniae and its relationship to virulence. Can J Microbiol 31, 472-478. Domenico, P., Tomas, J.M., Merino, S., Rubires, X., and Cunha, B.A. (1999). Surface antigen exposure by bismuth dimercaprol suppression of Klebsiella pneumoniae capsular polysaccharide. Infect Immun 67, 664-669. Erbing, C., Kenne, L., Lonngren, J., and Sutherland, I.W. (1976). Structural studies of the capsular polysaccharide from Klebsiella type 1. Carbohydr Res 50, 115-120. Fang, C.T., Chuang, Y.P., Shun, C.T., Chang, S.C., and Wang, J.T. (2004). A novel virulence gene in Klebsiella pneumoniae strains causing primary liver abscess and septic metastatic complications. J Exp Med 199, 697-705. Fang, C.T., Lai, S.Y., Yi, W.C., Hsueh, P.R., Liu, K.L., and Chang, S.C. (2007). Klebsiella pneumoniae genotype K1: an emerging pathogen that causes septic ocular or central nervous system complications from pyogenic liver abscess. Clin Infect Dis 45, 284-293. Fang, H.C. (2007). Identification of the role of rmpA in Klebsiella pneumoniae. In 微生 物學研究所 碩士論文 (台北國立台灣大學醫學院). Fung, C.P., Chang, F.Y., Lee, S.C., Hu, B.S., Kuo, B.I., Liu, C.Y., Ho, M., and Siu, L.K. (2002). A global emerging disease of Klebsiella pneumoniae liver abscess: is serotype K1 an important factor for complicated endophthalmitis? Gut 50, 420-424. Karama, E.M., Willermain, F., Janssens, X., Claus, M., Van den Wijngaert, S., Wang, J.T., Verougstraete, C., and Caspers, L. (2008). Endogenous endophthalmitis complicating Klebsiella pneumoniae liver abscess in Europe: case report. Int Ophthalmol 28, 111-113. Kau, A.L., Martin, S.M., Lyon, W., Hayes, E., Caparon, M.G., and Hultgren, S.J. (2005). Enterococcus faecalis tropism for the kidneys in the urinary tract of C57BL/6J mice. Infect Immun 73, 2461-2468. Ko, W.C., Paterson, D.L., Sagnimeni, A.J., Hansen, D.S., Von Gottberg, A., Mohapatra, S., Casellas, J.M., Goossens, H., Mulazimoglu, L., Trenholme, G., et al. (2002). 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Genomic heterogeneity in Klebsiella pneumoniae strains is associated with primary pyogenic liver abscess and metastatic infection. J Infect Dis 192, 117-128. Mizuta, K., Ohta, M., Mori, M., Hasegawa, T., Nakashima, I., and Kato, N. (1983). Virulence for mice of Klebsiella strains belonging to the O1 group: relationship to their capsular (K) types. Infect Immun 40, 56-61. Naito, T., Kawakami, T., Tsuda, M., Ebe, T., Sekiya, S., Isonuma, H., Matsumoto, T., and Watanabe, K. (1999). [A case of endophthalmitis and abscesses in the liver and the lung caused by Klebsiella pneumoniae]. Kansenshogaku Zasshi 73, 935-938. Nassif, X., Fournier, J.M., Arondel, J., and Sansonetti, P.J. (1989a). Mucoid phenotype of Klebsiella pneumoniae is a plasmid-encoded virulence factor. Infect Immun 57, 546-552. Nassif, X., Honore, N., Vasselon, T., Cole, S.T., and Sansonetti, P.J. (1989b). Positive control of colanic acid synthesis in Escherichia coli by rmpA and rmpB, two virulence-plasmid genes of Klebsiella pneumoniae. Mol Microbiol 3, 1349-1359. Pan, Y.J., Fang, H.C., Yang, H.C., Lin, T.L., Hsieh, P.F., Tsai, F.C., Keynan, Y., and Wang, J.T. (2008). Capsular polysaccharide synthesis regions in Klebsiella pneumoniae serotype K57 and a new capsular serotype. J Clin Microbiol 46, 2231-2240. Podschun, R., and Ullmann, U. (1998). Klebsiella spp. as nosocomial pathogens: epidemiology, taxonomy, typing methods, and pathogenicity factors. Clin Microbiol Rev 11, 589-603. Saccente, M. (1999). Klebsiella pneumoniae liver abscess, endophthalmitis, and meningitis in a man with newly recognized diabetes mellitus. Clin Infect Dis 29, 1570-1571. Sahly, H., Podschun, R., Oelschlaeger, T.A., Greiwe, M., Parolis, H., Hasty, D., Kekow, J., Ullmann, U., Ofek, I., and Sela, S. (2000). Capsule impedes adhesion to and invasion of epithelial cells by Klebsiella pneumoniae. Infect Immun 68, 6744-6749. Schembri, M.A., Dalsgaard, D., and Klemm, P. (2004). Capsule shields the function of short bacterial adhesins. J Bacteriol 186, 1249-1257. Simoons-Smit, A.M., Verweij-van Vught, A.M., and MacLaren, D.M. (1986). The role of K antigens as virulence factors in Klebsiella. J Med Microbiol 21, 133-137. 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., Wann, S.R., and Lin, H.H. (1998). Primary liver abscess due to Klebsiella pneumoniae in Taiwan. Clin Infect Dis 26, 1434-1438. Yu, W.L., Ko, W.C., Cheng, K.C., Lee, C.C., Lai, C.C., and Chuang, Y.C. (2008). Comparison of prevalence of virulence factors for Klebsiella pneumoniae liver abscesses between isolates with capsular K1/K2 and non-K1/K2 serotypes. Diagn Microbiol Infect Dis 62, 1-6. 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, 2231-2240. Podschun, R., and Ullmann, U. (1998). Klebsiella spp. as nosocomial pathogens: epidemiology, taxonomy, typing methods, and pathogenicity factors. Clin Microbiol Rev 11, 589-603. Saccente, M. (1999). Klebsiella pneumoniae liver abscess, endophthalmitis, and meningitis in a man with newly recognized diabetes mellitus. Clin Infect Dis 29, 1570-1571. Sahly, H., Podschun, R., Oelschlaeger, T.A., Greiwe, M., Parolis, H., Hasty, D., Kekow, J., Ullmann, U., Ofek, I., and Sela, S. (2000). Capsule impedes adhesion to and invasion of epithelial cells by Klebsiella pneumoniae. Infect Immun 68, 6744-6749. Schembri, M.A., Dalsgaard, D., and Klemm, P. (2004). Capsule shields the function of short bacterial adhesins. J Bacteriol 186, 1249-1257. Simoons-Smit, A.M., Verweij-van Vught, A.M., and MacLaren, D.M. (1986). The role of K antigens as virulence factors in Klebsiella. J Med Microbiol 21, 133-137. 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., Wann, S.R., and Lin, H.H. (1998). Primary liver abscess due to Klebsiella pneumoniae in Taiwan. Clin Infect Dis 26, 1434-1438. Yu, W.L., Ko, W.C., Cheng, K.C., Lee, C.C., Lai, C.C., and Chuang, Y.C. (2008). Comparison of prevalence of virulence factors for Klebsiella pneumoniae liver abscesses between isolates with capsular K1/K2 and non-K1/K2 serotypes. Diagn Microbiol Infect Dis 62, 1-6. 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, 2231-2240. Podschun, R., and Ullmann, U. (1998). Klebsiella spp. as nosocomial pathogens: epidemiology, taxonomy, typing methods, and pathogenicity factors. Clin Microbiol Rev 11, 589-603. Saccente, M. (1999). Klebsiella pneumoniae liver abscess, endophthalmitis, and meningitis in a man with newly recognized diabetes mellitus. Clin Infect Dis 29, 1570-1571. Sahly, H., Podschun, R., Oelschlaeger, T.A., Greiwe, M., Parolis, H., Hasty, D., Kekow, J., Ullmann, U., Ofek, I., and Sela, S. (2000). Capsule impedes adhesion to and invasion of epithelial cells by Klebsiella pneumoniae. Infect Immun 68, 6744-6749. Schembri, M.A., Dalsgaard, D., and Klemm, P. (2004). Capsule shields the function of short bacterial adhesins. J Bacteriol 186, 1249-1257. Simoons-Smit, A.M., Verweij-van Vught, A.M., and MacLaren, D.M. (1986). The role of K antigens as virulence factors in Klebsiella. J Med Microbiol 21, 133-137. 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., Wann, S.R., and Lin, H.H. (1998). Primary liver abscess due to Klebsiella pneumoniae in Taiwan. Clin Infect Dis 26, 1434-1438. Yu, W.L., Ko, W.C., Cheng, K.C., Lee, C.C., Lai, C.C., and Chuang, Y.C. (2008). Comparison of prevalence of virulence factors for Klebsiella pneumoniae liver abscesses between isolates with capsular K1/K2 and non-K1/K2 serotypes. Diagn Microbiol Infect Dis 62, 1-6. 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, 2231-2240. Podschun, R., and Ullmann, U. (1998). Klebsiella spp. as nosocomial pathogens: epidemiology, taxonomy, typing methods, and pathogenicity factors. Clin Microbiol Rev 11, 589-603. Saccente, M. (1999). Klebsiella pneumoniae liver abscess, endophthalmitis, and meningitis in a man with newly recognized diabetes mellitus. Clin Infect Dis 29, 1570-1571. Sahly, H., Podschun, R., Oelschlaeger, T.A., Greiwe, M., Parolis, H., Hasty, D., Kekow, J., Ullmann, U., Ofek, I., and Sela, S. (2000). Capsule impedes adhesion to and invasion of epithelial cells by Klebsiella pneumoniae. Infect Immun 68, 6744-6749. Schembri, M.A., Dalsgaard, D., and Klemm, P. (2004). Capsule shields the function of short bacterial adhesins. J Bacteriol 186, 1249-1257. Simoons-Smit, A.M., Verweij-van Vught, A.M., and MacLaren, D.M. (1986). The role of K antigens as virulence factors in Klebsiella. J Med Microbiol 21, 133-137. 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., Wann, S.R., and Lin, H.H. (1998). Primary liver abscess due to Klebsiella pneumoniae in Taiwan. Clin Infect Dis 26, 1434-1438. Yu, W.L., Ko, W.C., Cheng, K.C., Lee, C.C., Lai, C.C., and Chuang, Y.C. (2008). Comparison of prevalence of virulence factors for Klebsiella pneumoniae liver abscesses between isolates with capsular K1/K2 and non-K1/K2 serotypes. Diagn Microbiol Infect Dis 62, 1-6. 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. | |
dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/45379 | - |
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,經由社區感染,會引起典型的臨床表現—原發性肝膿瘍(community-acquired pyogenic liver abscess,PLA)合併菌血症,莢膜為菌株之主要致病因子之一。rmpA/A2基因能促進菌株的莢膜表現量進而影響黏性型態與致病力,在從台大醫院收集的引發原發性肝膿瘍菌株中rmpA/A2之比例為100%(60/60),而在非組織侵襲性菌株中為22%(7/32)。在本實驗室代表性菌株NTUH-K2044中有三套rmpA/A2基因,分析三套rmpA/A2基因在臨床菌株中之分佈,發現以質體pK2044上的rmpA及rmpA2基因分布比例較高。以NTUH-K2044建構染色體上rmpA基因剔除株(Δ chromosome rmpA)、質體pK2044上rmpA基因剔除株(Δ plasmid rmpA)與質體pK2044上截斷形式的rmpA2基因剔除株(Δ plasmid rmpA2),結果顯示Δ chromosome rmpA突變株對黏性、莢膜生合成量及其基因組表現無影響;Δ plasmid rmpA突變株能明顯降低黏性型態及莢膜生合成基因組表現;Δ plasmid rmpA2突變株之莢膜生合成基因組表現有些微上升,但不影響黏性型態及莢膜生合成量。而在以腹膜腔注射及胃內感染方式探討對小鼠致病力的結果中,Δ plasmid rmpA突變株與野生株亦無明顯差異,然而在小鼠體內競爭實驗中,則發現Δ plasmid rmpA突變株在肝臟及脾臟的散佈生長情形明顯較野生株強。另外,從三株基因剔除株中分析三套rmpA/A2基因的表現量結果得知,只有在失去質體 pK2044 rmpA基因時,會降低染色體rmpA及質體pK2044 rmpA2的表現量;染色體rmpA基因對質體 pK2044 rmpA及rmpA2基因的表現量影響不大;質體 pK2044 上截斷的rmpA2則有些微抑制另兩套rmpA基因的表現量。染色體rmpA基因與質體 pK2044 rmpA基因序列相同度為92%,但只有質體 pK2044 rmpA基因會影響莢膜生合成表現,將野生株中的質體 pK2044 rmpA置換為染色體rmpA後,莢膜基因組表現量有明顯下降。因此,由本實驗結果可知,在NTUH-K2044菌株內三套rmpA/A2中,只有破壞質體 pK2044rmpA會降低莢膜表現量及黏性,並且增強菌株於小鼠體內器官中散佈生長的情形;而造成染色體rmpA及質體 pK2044 rmpA兩者在調控黏性功能上的不同源於兩者之基因序列不同。 | zh_TW |
dc.description.abstract | There are three copies of rmpA/A2 (regulator of the mucoid phenotype A) genes located in the chromosome or large plasmid of Klebsiella pneumoniae NTUH-K2044 strain. This gene has been reported to be involved in the biosynthesis of capsular polysaccharide. A copy of rmpA is located on the chromosome. A rmpA and a truncated rmpA2 are located at the pK2044 plasmid of NTUH-K2044. In order to study the specific biological function of these three rmpA/A2 genes, we generated three rmpA/A2 single deletion mutants. We found that the deletion of plasmid rmpA resulted in decreases of K1 CPS-related gene expression and colony mucoviscosity. Comparing with the wild-type strain, the chromosome rmpA deletion mutant showed no significant difference in K1 CPS-related gene expression and colony mucoviscosity, whereas the plasmid rmpA2 mutant had an increase in the K1 CPS-related gene expression but not in the K1 CPS synthesis. In in vivo virulence analysis, we used intraperitoneal inoculation infection in BALB/c mice model to perform in vivo competition assay. The results revealed that colonization ability of plasmid rmpA mutant was stronger than that of wild type in liver, and spleen. This implied that the decrease of the K1-CPS synthesis of K. pneumoniae in Δplasmid rmpA mutant might contribute to facilitate in vivo colonization of K. pneumoniae. In addition, chromosome rmpA and plasmid rmpA2 mRNA expression was decreased in the plasmid rmpA2 mutant compared than the wild-type strain. Chromosome rmpA gene and plasmid rmpA gene shared a 92% identity on DNA sequence. However, only plasmid rmpA was capable of up-regulating the expression of cps gene. While plasmid rmpA was replaced by chromosome rmpA in NTUH-K2044, the expression of cps genes decreased.
We concluded that plasmid rmpA had the ability to enhance CPS synthesis, but chromosome rmpA and plasmid rmpA2 had no influence on CPS synthesis. Moreover, destruction of plasmid rmpA promoted bacterial colonization in liver and spleen.The distinct ability of regulating cps region between chromosome rmpA and plasmid rmpA is the result of their different sequences. | en |
dc.description.provenance | Made available in DSpace on 2021-06-15T04:17:02Z (GMT). No. of bitstreams: 1 ntu-98-R96445125-1.pdf: 1270780 bytes, checksum: cfb20d2bac6ea7a3e7b4b374e3cf0e9b (MD5) Previous issue date: 2009 | en |
dc.description.tableofcontents | 目錄
口試委員會審定書 ........................................................................................................... I 致謝 ................................................................................................................................. II 中文摘要 ......................................................................................................................... III 英文摘要 .......................................................................................................................... V 第一章、緒論 ................................................................................................................... 1 第二章 材料與方法 ......................................................................................................... 6 2.1細菌菌株與質體 .....................................................................................................6 2.2聚合酶連鎖反應(polymerase chain reaction, PCR) ..........................................6 2.3建構克雷伯氏肺炎桿菌rmpA/A2基因剔除株 .....................................................7 2.4萃取克雷伯氏肺炎桿菌基因體DNA(genomic DNA) ....................................7 2.5南方轉漬法(Sourthern blot) ..............................................................................8 2.6萃取克雷伯氏肺炎桿菌total RNA ........................................................................9 2.7反轉錄定量聚合酶鏈鎖反應(RT-qPCR) .........................................................9 2.8拉絲測試(String test) .......................................................................................10 2.9低速離心試驗 .......................................................................................................11 2.10濃硫酸測定醣類試驗 .........................................................................................11 2.11莢膜血清型分型(Serotyping) ........................................................................11 2.12 Δ plasmid rmpA突變株之染色體上互補菌株(chromosome complementation) ................................................................................................................................... 12 2.13血清抗性試驗(Serum resistance assay) .........................................................12 2.14動物實驗 .............................................................................................................13 2.15建構異位互補質體 .............................................................................................14 第三章、結果 ................................................................................................................. 15 3.1觀察三套rmpA/A2基因在臨床菌株中的分佈 ...................................................15 3.2在克雷伯氏肺炎桿菌NTUH-K2044菌株中,建構三株突變株:Δ chromosome rmpA,Δ plasmid rmpA及Δ plasmid rmpA2 ............................................................16 3.3分析在三株rmpA/A2基因剔除株中莢膜生合成基因組的表現 .......................17 3.4觀察三株rmpA/A2基因剔除株的黏性、莢膜生合成量及K1血清型 ............18 3.5建構Δ plasmid rmpA染色體上補回菌株(chromosome complementation)並分析其特性 .....................................................................................................................19 3.6 探討NTUH-K2044與Δ plasmid rmpA突變株對血清抗性及對小鼠的致病力 ....................................................................................................................................20 3.7比較野生株、三株rmpA/A2基因剔除株、Δ plasmid rmpA染色體上補回菌株中三套rmpA/A2 RNA表現量 ....................................................................................22 3.8比較NTUH-K2044野生菌株中染色體 rmpA及質體 pK2044 rmpA之表現量 ....................................................................................................................................23 3.9建構一株將野生株的質體pK2044 rmpA基因置換成染色體rmpA基因序列的突變株,分析其對莢膜生合成量與其基因表現量的影響 .....................................24 第四章、討論 ................................................................................................................. 26 表目錄 表一、研究使用之菌株及載體 .....................................................................................33 表二、實驗中使用到的引子 .........................................................................................34 表三、以PCR檢測PLA與non-invasive strains之三套rmpA/A2基因及其血清型的分佈 ................................................................................................................................36 表四、以拉絲測驗及沉澱離心試驗測定NTUH-K2044野生株、三株rmpA/A2基因剔除株、及Δ plasmid rmpA染色體上互補菌株之黏性型態 .....................................37 表五、以腹膜腔注射的方式,比較NTUH-K2044野生株及Δ plasmid rmpA突變株對於小鼠的致病力 .........................................................................................................38 表六、以體內競爭實驗(in vivo competition assay),利用PlacZ基因剔除株作為野生株代表,檢視Δplasmid rmpA突變株在小鼠中散佈生長(colonization)情形 ............39 圖目錄 圖一、利用Southern blot 鑑定Δ chromosome rmpA,Δ plasmid rmpA突變株中,染色體rmpA基因及質體pK2044 rmpA基因是否存在。 ............................................. 40 圖二、利用RT-qPCR來比較在NTUH-K2044野生株、三株rmpA/A2基因剔除株以及Δ plasmid rmpA染色體互補菌株中galF(A)、wzc(B)基因的RNA表現量。 ........................................................................................................................................41 圖三、利用濃硫酸定量糖類試驗比較在NTUH-K2044野生株、三株rmpA/A2基因剔除株、以及Δ plasmid rmpA染色體上互補菌株的莢膜生合成量。 .....................42 圖四、利用雙向免疫擴散法來偵測三株rmpA/A2基因剔除株的K1莢膜多醣體。 ........................................................................................................................................43 圖五、利用RT-qPCR分析Δ plasmid rmpA染色體上互補菌株中質體pK2044 rmpA基因表現量。 .................................................................................................................44 圖六、利用血清抗性試驗來比較NTUH-K2044野生株與Δ plasmid rmpA突變株之血清抗性。 .....................................................................................................................45 圖七、以胃內感染方式,比較NTUH-K2044野生株及Δ plasmid rmpA突變株對於小鼠的致病力。 .............................................................................................................46 圖八、利用RT-qPCR 分析在NTUH-K2044野生株、三株rmpA/A2基因剔除株、及Δ plasmid rmpA染色體上互補菌株中的染色體rmpA(A)、質體pK2044 rmpA(B)、與質體pK2044 rmpA2(C)基因的RNA表現量。 ....................................47 圖九、建構染色體rmpA與質體pK2044 rmpA載體的RT-qPCR的標準曲線。 ...48 圖十、利用RT-qPCR 分析在NTUH-K2044野生株中染色體rmpA與質體pK2044 rmpA基因的RNA表現量。 .........................................................................................49 圖十一、比較NTUH-K2044野生株與PrmpA→CrmpA突變株內莢膜生合成基因galF(A)、wzc(B)的RNA表現量與莢膜生合成量(C)。 ....................................50 | |
dc.language.iso | zh-TW | |
dc.title | 克雷伯氏肺炎桿菌中三套rmpA/A2基因之功能分析 | zh_TW |
dc.title | Functional characterization of three rmpA/A2 loci in Klebsiella pneumoniae | en |
dc.type | Thesis | |
dc.date.schoolyear | 98-1 | |
dc.description.degree | 碩士 | |
dc.contributor.oralexamcommittee | 陳美如(Mei-Ru Chen),林稚容(Tzu-Lung Lin) | |
dc.subject.keyword | 克雷伯氏肺炎桿菌,rmpA/A2基因,莢膜,cps gene cluster, | zh_TW |
dc.subject.keyword | Klebsiella pneumoniae,rmpA/A2,capsule,,cps gene cluster, | en |
dc.relation.page | 54 | |
dc.rights.note | 有償授權 | |
dc.date.accepted | 2009-12-28 | |
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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