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完整後設資料紀錄
DC 欄位 | 值 | 語言 |
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dc.contributor.advisor | 王錦堂(Jin-Town Wang) | |
dc.contributor.author | Hsiao-Hsuan Lin | en |
dc.contributor.author | 林筱萱 | zh_TW |
dc.date.accessioned | 2021-06-15T06:46:01Z | - |
dc.date.available | 2021-12-31 | |
dc.date.copyright | 2011-10-07 | |
dc.date.issued | 2011 | |
dc.date.submitted | 2011-06-23 | |
dc.identifier.citation | Adams, M.H., and B.H. Park. 1956. An enzyme produced by a phage-host cell system. II. The properties of the polysaccharide depolymerase. Virology 2:719-736.
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Clarke, B.R., F. Esumeh, and I.S. Roberts. 2000. Cloning, expression, and purification of the K5 capsular polysaccharide lyase (KflA) from coliphage K5A: evidence for two distinct K5 lyase enzymes. J Bacteriol 182:3761-3766. Drulis-Kawa, Z., P. Mackiewicz, A. Kesik-Szeloch, E. Maciaszczyk-Dziubinska, B. Weber-Dabrowska, A. Dorotkiewicz-Jach, D. Augustyniak, G. Majkowska-Skrobek, T. Bocer, J. Empel, and A.M. Kropinski. 2011. Isolation and characterisation of KP34--a novel phiKMV-like bacteriophage for Klebsiella pneumoniae. Appl Microbiol Biotechnol 90:1333-1345. Durlakowa, I., Z. Lachowicz, and S. Slopek. 1967. Serologic characterization of Klebsiella bacilli on the basis of properties of the capsular antigens. Arch Immunol Ther Exp (Warsz) 15:497-504. Dutton, G.G., H. Parolis, J.P. Joseleau, and M.F. Marais. 1986. The use of bacteriophage depolymerization in the structural investigation of the capsular polysaccharide from Klebsiella serotype K3. Carbohydr Res 149:411-423. DW, S.J.a.R. 2001. Molecular Cloning: A Laboratory Manual (Third). York, New Fang, C.T., S.Y. Lai, W.C. Yi, P.R. Hsueh, K.L. Liu, and S.C. Chang. 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. Fung, C.P., F.Y. Chang, S.C. Lee, B.S. Hu, B.I. Kuo, C.Y. Liu, M. Ho, and L.K. Siu. 2002. A global emerging disease of Klebsiella pneumoniae liver abscess: is serotype K1 an important factor for complicated endophthalmitis? Gut 50:420-424. Fung, C.P., B.S. Hu, F.Y. Chang, S.C. Lee, B.I. Kuo, M. Ho, L.K. Siu, and C.Y. Liu. 2000. A 5-year study of the seroepidemiology of Klebsiella pneumoniae: high prevalence of capsular serotype K1 in Taiwan and implication for vaccine efficacy. J Infect Dis 181:2075-2079. Gaston, M.A., B.A. Ayling-Smith, and T.L. Pitt. 1987. New bacteriophage typing scheme for subdivision of the frequent capsular serotypes of Klebsiella spp. J Clin Microbiol 25:1228-1232. Hackland, P.L., H. Parolis, and L.A. Parolis. 1988. A structural investigation of the capsular polysaccharide of Klebsiella K69. Carbohydr Res 172:209-216. Hyman, P., and S.T. Abedon. 2010. Bacteriophage host range and bacterial resistance. Adv Appl Microbiol 70:217-248. Jenney, A.W., A. Clements, J.L. Farn, O.L. Wijburg, A. McGlinchey, D.W. Spelman, T.L. Pitt, M.E. Kaufmann, L. Liolios, M.B. Moloney, S.L. Wesselingh, and R.A. Strugnell. 2006. Seroepidemiology of Klebsiella pneumoniae in an Australian Tertiary Hospital and its implications for vaccine development. J Clin Microbiol 44:102-107. Ko, W.C., D.L. Paterson, A.J. Sagnimeni, D.S. Hansen, A. Von Gottberg, S. Mohapatra, J.M. Casellas, H. Goossens, L. Mulazimoglu, G. Trenholme, K.P. Klugman, J.G. McCormack, and V.L. Yu. 2002. Community-acquired Klebsiella pneumoniae bacteremia: global differences in clinical patterns. Emerg Infect Dis 8:160-166. Krwawicz, J., A. Czajkowska, M. Felczak, and I. Pietrzykowska. 2003. UV- and MMS-induced mutagenesis of lambdaO(am)8 phage under nonpermissive conditions for phage DNA replication. Acta Biochim Pol 50:921-939. Kumari, S., K. Harjai, and S. Chhibber. 2010. Isolation and characterization of Klebsiella pneumoniae specific bacteriophages from sewage samples. Folia Microbiol (Praha) 55:221-227. Labrie, S.J., J.E. Samson, and S. Moineau. 2010. Bacteriophage resistance mechanisms. Nat Rev Microbiol 8:317-327. Lindberg, B., F. Lindh, J. Lonngren, and I.W. Sutherland. 1979. Structural studies of the capsular polysaccharide of Klebsiella type 30. Carbohydr Res 76:281-284. Link, A.J., D. Phillips, and G.M. Church. 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., D.L. Cheng, and C.L. Lin. 1986. Klebsiella pneumoniae liver abscess associated with septic endophthalmitis. Arch Intern Med 146:1913-1916. Palfreyman, J.M. 1978. Klebsiella serotyping by counter-current immunoelectrophoresis. J Hyg (Lond) 81:219-225. Pan, Y.J., H.C. Fang, H.C. Yang, T.L. Lin, P.F. Hsieh, F.C. Tsai, Y. Keynan, and J.T. Wang. 2008. Capsular polysaccharide synthesis regions in Klebsiella pneumoniae serotype K57 and a new capsular serotype. J Clin Microbiol 46:2231-2240. Park, B.H. 1956. An enzyme produced by a phage-host cell system. I. The properties of a Klebsiella phage. Virology 2:711-718. Pelkonen, S., J. Aalto, and J. Finne. 1992. Differential activities of bacteriophage depolymerase on bacterial polysaccharide: binding is essential but degradation is inhibitory in phage infection of K1-defective Escherichia coli. J Bacteriol 174:7757-7761. Pieroni, P. 1996. Application of Bacteriophage Typing to Klebsiella pneumoniae. In Saskatchewan, Canada. Pieroni, P., R.P. Rennie, B. Ziola, and H.G. Deneer. 1994. The use of bacteriophages to differentiate serologically cross-reactive isolates of Klebsiella pneumoniae. J Med Microbiol 41:423-429. Podschun, R., and U. Ullmann. 1998. Klebsiella spp. as nosocomial pathogens: epidemiology, taxonomy, typing methods, and pathogenicity factors. Clin Microbiol Rev 11:589-603. Qiagen. 1999. Qiagen Lambda Hamdbook. Rakhuba, D.V., E.I. Kolomiets, E.S. Dey, and G.I. Novik. 2010. Bacteriophage receptors, mechanisms of phage adsorption and penetration into host cell. Pol J Microbiol 59:145-155. Ravenscroft, N., L.A. Parolis, and H. Parolis. 1994. Bacteriophage degradation of Klebsiella K30 capsular polysaccharide. An NMR investigation of the 3,4-pyruvated galactose-containing repeating oligosaccharide. Carbohydr Res 254:333-340. Rieger-Hug, D., and S. Stirm. 1981. Comparative study of host capsule depolymerases associated with Klebsiella bacteriophages. Virology 113:363-378. Sahly, H., and R. Podschun. 1997. Clinical, bacteriological, and serological aspects of Klebsiella infections and their spondylarthropathic sequelae. Clin Diagn Lab Immunol 4:393-399. Scholl, D., S. Rogers, S. Adhya, and C.R. Merril. 2001. Bacteriophage K1-5 encodes two different tail fiber proteins, allowing it to infect and replicate on both K1 and K5 strains of Escherichia coli. J Virol 75:2509-2515. Sechter, I., F. Mestre, and D.S. Hansen. 2000. Twenty-three years of Klebsiella phage typing: a review of phage typing of 12 clusters of nosocomial infections, and a comparison of phage typing with K serotyping. Clin Microbiol Infect 6:233-238. Tsay, R.W., L.K. Siu, C.P. Fung, and F.Y. Chang. 2002. Characteristics of bacteremia between community-acquired and nosocomial Klebsiella pneumoniae infection: risk factor for mortality and the impact of capsular serotypes as a herald for community-acquired infection. Arch Intern Med 162:1021-1027. | |
dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/48091 | - |
dc.description.abstract | 克雷伯氏肺炎桿菌是造成院內與社區感染的常見菌株,莢膜為其重要的致病因子之一,莢膜又被稱為K抗原(K antigen),莢膜分型一直以來都是流行病學了解菌株與疾病之間的唯一方法,也可能有助於疫苗的研發。傳統以來一直都是利用血清法來進行莢膜分型,然而血清分型法敏感度與專一性都不佳,因此先前有研究利用噬菌體來進行克雷伯氏肺炎桿菌部分莢膜分型,發現可感染克雷伯氏肺炎桿菌的噬菌體,尾端通常帶有莢膜醣類分解酵素(Capsule depolymerase),然而先前研究並無核酸序列或病毒檢體留存。因此本實驗室重新研究使用噬菌體所帶有的莢膜醣類分解酵素來區分莢膜型的可能,我們在分離對莢膜具有專一性噬菌體過程中,發現有許多噬菌體能感染多種莢膜型的克雷伯氏肺炎桿菌。因此本論文研究目的希望找出噬菌體能感染多種莢膜型的原因。實驗結果發現,從水源分離出來能夠感染K5莢膜型的噬菌體,共有四株ΦK5-1、ΦK5-2、ΦK5-3(A44)、ΦK5-4(A44)。而其中ΦK5-2與ΦK5-4(A44)並不具專一性,能感染多種莢膜型。此外,透過高通量定序,完成了ΦK5-2與ΦK5-4(A44)基因體定序,經過基因序列的比對,發現ΦK5-2與ΦK5-4(A44),帶有多種莢膜醣類分解酵素使噬菌體能夠感染多種莢膜型。從這兩株噬菌體,共得到了四種可能的酵素基因序列,經由選殖表現載體進行蛋白質表現與純化。實驗結果證實這些基因產物為K8莢膜醣類分解酵素K5-4(A44)_ORF23、K5莢膜醣類分解酵素K5-4(A44)_ORF24與K30/K69莢膜醣類分解酵素K5-2_ORF26。另外,利用化學藥劑產生的突變株Φ2-K5-2,證實了失去莢膜醣類分解酵素,會影響噬菌體的宿主範圍。而這些莢膜醣類分解酵素也具有專一性,利用這些莢膜醣類分解酵素進行分型,其敏感度比噬菌體本身效果來得好。未來期望能夠找出所有莢膜醣類分解酵素基因,建立完成出一套新的克雷伯氏肺炎桿菌莢膜分型系統。 | zh_TW |
dc.description.abstract | Klebsiella pneumoniae is a common cause of community-acquired and nosocomial infections. It has polysaccharide capsule that is a major pathogenicity factor. Capsule was also called K antigen. Determination of the K type has long been the preferred method for the investigation of epidemiological relationships among strains and was also possibly helpful to the vaccine development. In the past, serotyping can be employed to determine the K type. Moreover, the anti-sera had limited specificity and sensitivity in the previous reports. Therefore, previous studies reported that use of bacteriophages can determine part of capsule types. They found that bacteriophages infected K. pneumoniae often carried the capsule depolymerase in the tail. However, these phages were no longer available and there was no sequencing result in previous studies. Therefore, our lab tried to study capsule depolymerase which come from capsular-type-specific bacteriophages and established a new method for capsular typing. Capsular-type-specific bacteriophages for K. pneumonia were isolated in our lab. But we also found that many bacteriophages can infect more than one capsular types of K. pneumoniae. Therefore, the aim of this research is to determine why the bacteriophage can infect more than one capsular types of K. pneumoniae. We isolate four bacteriophages which could infect K. pneumoniae strains with K5 capsule type from untreated water, ΦK5-1, Φ K5-2, ΦK5-3(A44) and ΦK5-4(A44). ΦK5-2 and ΦK5-4(A44) infected more than one capsular type strains. By high-throughput DNA sequencing, we completed genome sequence of ΦK5-2 and ΦK5-4(A44). By comparison with NCBI BLAST databases, we found bacteriophage encodes two different glycosidases, allowing it to infect two capsule type strains. From these bacteriophages, we got four glycosidases sequence. We expressed and purified these glycosidases to demonstrate their function. In our results, K5-4(A44)_ORF23 was a glycosidase to digest K8 capsule. K5-4(A44)_ORF24 was a glycosidase to digest K5 capsule. K5-2_ORF26 was a glycosidase to digest K30 and K69 capsules. By mutagenesis, we found that the mutant Φ2-K5-2 which lost glycosidase activity changed its host range. Capsular typing by these glycosidases was more specific and sensitive than by bacteriophages. In the future, we hope a glycosidases typing system covered all capsular types will be established. | en |
dc.description.provenance | Made available in DSpace on 2021-06-15T06:46:01Z (GMT). No. of bitstreams: 1 ntu-100-R98445102-1.pdf: 2618470 bytes, checksum: 3adcf0fe738f1ba93869359ad8ee4f33 (MD5) Previous issue date: 2011 | en |
dc.description.tableofcontents | 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 第二章、材料與方法.........................................5 1. 實驗菌株.........................................5 2. 質體與抗生素.....................................5 3. 尋找及分離克雷伯氏肺炎桿菌莢膜型K5的噬菌體.......5 4. 塗點試驗(spot test) .............................7 5. 噬菌體溶菌斑效價分析(plaque assay) ..............7 6. 噬菌體增殖(phage amplification) .................8 7. 噬菌體純化與DNA萃取..............................8 8. 限制性核酸內切酶實驗.............................10 9. 噬菌體DNA高通量定序.............................10 10. 細菌吸附試驗....................................10 11. 噬菌體的序列分析................................10 12. 隨機突變試驗(Random mutagenesis)................11 13. 聚合酶鍊反應PCR(polymerase chain reaction)......12 14. 建構克雷伯氏肺炎桿菌莢膜型K5莢膜合成基因突變株..13 15. 噬菌體的莢膜醣類分解酵素蛋白質表現與純化........14 16. 十二烷基硫酸鈉-聚丙烯酰胺凝膠電泳(SDS-PAGE) ....16 17. 西方轉漬(Western blot) .........................18 第三章、實驗結果..........................................20 1. 尋找及分離克雷伯氏肺炎桿菌K5莢膜型之噬菌體......20 2. 萃取噬菌體之基因體..............................20 3. 噬菌體宿主範圍及其感染能力......................20 4. K5噬菌體感染宿主與宿主莢膜存在有關..............21 5. 隨機突變改變噬菌體宿主範圍......................21 6. 噬菌體基因體定序分析............................22 7. 莢膜醣類分解酵素的蛋白質純化....................23 8. 莢膜醣類分解酵素的功能測試......................23 9. 比較莢膜醣類分解酵素與噬菌體對於同莢膜型不同菌株的敏感度測試................................................24 第四章、討論與總結........................................25 第五章、參考文獻..........................................59 | |
dc.language.iso | zh-TW | |
dc.title | 克雷伯氏肺炎桿菌K5莢膜型之噬菌體的醣解酵素活性與宿主專一性 | zh_TW |
dc.title | Glycosidase activity and host specificity of Bacteriophage K5 of Klebsiella pneumoniae | en |
dc.type | Thesis | |
dc.date.schoolyear | 99-2 | |
dc.description.degree | 碩士 | |
dc.contributor.oralexamcommittee | 葉秀慧,楊宏志,林稚容 | |
dc.subject.keyword | 克雷伯氏肺炎桿菌,噬菌體,莢膜分解酵素, | zh_TW |
dc.subject.keyword | Klebsiella pneumonia,bacteriophage,capsule depolymerase, | en |
dc.relation.page | 62 | |
dc.rights.note | 有償授權 | |
dc.date.accepted | 2011-06-23 | |
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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