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http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/25272完整後設資料紀錄
| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 劉俊民 | |
| dc.contributor.author | Chao-Chun Fu | en |
| dc.contributor.author | 傅超群 | zh_TW |
| dc.date.accessioned | 2021-06-08T06:07:18Z | - |
| dc.date.copyright | 2007-07-23 | |
| dc.date.issued | 2007 | |
| dc.date.submitted | 2007-07-19 | |
| dc.identifier.citation | 陳素漪. 2002. 嗜水性產氣單胞桿菌Aeromonas hydrophila 轉形方法及分泌系統相關基因之選殖. 國立台灣大學農業化學研究所碩士論文.
翁晨耀. 2005. 嗜水性產氣單胞桿菌Aeromonas hydrophila 質體之探討與表現載體之建構. 國立台灣大學微生物與生化學研究所碩士論文. 李銘亮. 2002. 微生物生理學, 第一版., 台北：藝軒. 柯勇. 2003. 現代微生物學, 第一版, 第 13、14 章., 台北：藝軒. Actis, L. A., Tolmasky, M. E., and Crosa, J. H. 1999. Bacterial plasmids: replication of extrachromosomal genetic elements encoding resistance to antimicrobial compounds. Front. Biosci., 4:D43-62. Bhattacharya S. K., Dudey A. K. 1995. Metabolic burden as reflected by maintenance coefficient of recombinant Escherichia coli overexpressing target gene. Biotechnol. Lett., 17:1155–1160. Bouet, J. Y., Rech, J., Egloff, S., Biek, D. P., and Lane D. 2005. Probing plasmid partition with centromere-based incompatibility. Mol. Microbiol., 55(2):511-25. Boyd, J., Williams, J., Curtis, B., Kozera, C., Singh, R.,and Reith, M. 2003. Three small, cryptic plasmids from Aeromonas salmonicida subsp. salmonicida A449. Plasmid, 50(2):131-144. Chattoraj, D. K. 2000. Control of plasmid DNA replication by iterons: no longer paradoxical. Mol. Microbiol., 37(3):467-476. Cheah, U. E., Weigand, W. A., and Stark, B. C. 1987. Effects of recombinant plasmid size on cellular processes in Escherichia coli. Plasmid, 18:127-134. Clewell, D. B., Francia, M. V., Flannagan, S. E., and An, F. Y. 2002. Enterococcal plasmid transfer: sex pheromones, transfer origins, relaxases, and the Staphylococcus aureus issue. Plasmid, 48(3):193-201. del Solar, G. and Espinosa, M. 2000. Plasmid copy number control: an ever-growing story. Mol. Microbial., 37:492-500. del Solar, G., Giraldo, R., Ruiz-Echevarría, M. J., Espinosa, M., and Díaz-Orejas R. 1998. Replication and control of circular bacterial plasmids. Microbiol. Mol. Biol. Rev., 62:434-464. Gerhardt, P., Murray, R. G. E., Wood, W. A., Krieg, N. R. 1994. Methods for general and molecular bacteriology, p.383-384, Washington DC : American Society for Microbiology. Gotfredsen, M. and Gerdes, K. 1998. The Escherichia coli relBE genes belong to a new toxin-antitxoxin gene family. Mol. microbial., 29(4):1065-1076. Guerry, P. and Colwell, R. R. 1977. Isolation of cryptic plasmid deoxyribonucleic acid from Kanagawa-positive Strains of Vibrio parahaemolyticus. Infect. Immun., 16(1):328-334. Hooper, D. C., Wolfson, J. S., McHugh, G. L., Swartz, M. D., TUNG, C., and Swartz, M. N. 1984. Elimination of plasmid pMG110 from Escherichia coli by novobiocin and other inhibitors of DNA gyrase. Antimicrob. Agents. Chemother., 25(5): 586-590. Hayes, F. 2003. Toxins-Antitoxins: Plasmid Maintenance, Programmed Cell Death, and Cell Cycle Arrest Science, 301(5639):1496-1499. Kües, U. and Stahl, U. 1989. Replication of plasmids in gram-negative bacteria. Microbiol. Rev., 53(4): 491–516. Larry, S. and Wendy, C. 2003. Molecular genetics of bacteria, 2nd ed., Washington DC: ASM Press published. Mansi El-Mansia, Andersona, K. J., Inchea, C. A., Knowlesa, L. K. and Plattb, D. J. 2000. Isolation and curing of the Klebsiella pneumoniae large indigenous plasmid using sodium dodecyl sulphate. Res. Microbiol., 151:201-208. Meijer, W. J. J., Venema, G. and Bron, S. 1995. Characterization of single strand origins of cryptic rolling-circle plasmids from Bacillus subtilis. Nucleic acids Res., 23:612-619. Moscoso, M., del Solar, G., and Espinosa, M. 1995. Specific nicking-closing activity of the initiator of replication protein RepB of plasmid pMV158 on supercoiled or single-stranded DNA. J. Biol. Chem., 270:3772-3779 Novick, R. P. 1987. Plasmid incompatibility. Microbiol. Rev., 51(4):381–395. Paulsson, J. and Chattoraj, D. K. 2006. Origin inactivation in bacterial DNA replication control. Mol. Microbial., 61:9-15. Pillidge, C. J., Cambourn, W. M., and Pearce, L. E. 1996. Nucleotide sequence and analysis of pWC1, A pC194-type rolling circle replicon in Lactococcus lactis. Plasmid, 35:131-140. Ruiz-Barba J. L., Piard J. C., and Jimenez-Diaz R. 1991. Plasmid profiles and curing of plasmids in Lactobacillus plantarum strains isolated from green olive fermentations. J. Appl. Bacteriol., 71(5):417-421. Saleem, A. K. 2005. Plasmid rolling-circle replication: highlights of two decades of research. Plasmid, 53: 126-136. Scott, J. R. 1984. Regulation of Plasmid Replication. Microbiol. Rev., 48:1-23. Son, R., Rusul, G., Sahilah, A. M., Zainuri, A., Raha, A. R., Salmah, I. 1997. Antibiotic resistance and plasmid profile of Aeromonas hydrophila isolates from cultured fish, Telapia (Telapia mossambica). Lett. Appl. Microbiol., 24:479-482 Sukupolvi, S. and O'Connor, C. D. 1990. TraT lipoprotein, a plasmid-specified mediator of interactions between gram-negative bacteria and their environment. Microbiol. Rev., 54(4):331-341. Wang, Z., Yuan, Z., and Ulrich R. H. 2004. Processing of plasmid DNA with ColE1-like replication origin. Plasmid, 51:149-161. | |
| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/25272 | - |
| dc.description.abstract | 嗜水性產氣單胞桿菌為革蘭氏陰性、兼性厭氧性菌，廣泛的存在於各類水域之中，一般被視為伺機性病原菌，對於人類及動物均有致病力，並為重要之水產食品病原菌。但目前為止本菌仍然缺乏適當的載體系統可研究其基因功能，所以本研究的目標是期望由 Aeromonas 屬分離內生質體，分析其質體相關之複製等性質，以建構成具有複製區域之選殖載體，提供本菌分子生物研究之利用。本研究中由實驗室所保存之 Aeromonas 環境及臨床株中測定其中存在之質體，其中有 6 株帶有質體，菌株分別為 A. hydrophila 25、A. hydrophila 35、A. hydrophila 132、A. hydrophila 163、A. hydrophila 191、A. hydrophila 1028，從中挑選A. hydrophila 35、A. hydrophila 163 株之質體 pAH35、pAH163 進行全基因組 DNA 定序，將所得序列以 WU-BlastN 與核酸資料庫進行比對，在 pAH35 上找到與存在 E. coli 之 ColE1 質體之 RNA II 及 RNA I 相似之序列，顯示其可能為 ColE1-type 之質體。另外 pAH35 上亦找到與 E. coli ColE2-type 質體類似之 RNA I 及 Rep 蛋白質，顯示其亦可能為 ColE2-type 之質體，但目前尚未明瞭 pAH35 以何種機制複製。pAH163之序列比對出與複製相關之 Rep 蛋白質之 coding region，因此有可能是以 rolling circle 的方式複製。以 SDS、EtBr 及提高培養溫度之方式進行質體剔除後，由 A. hydrophila 35及 A. hydrophila 163分別得到 1 株 (AH35c1) 及 4 株 (AH163c1~4) 質體剔除株，分析其生理特性大致與野生株相近。為探討複製起始區之確實位置及計算質體之備份數，分別再以野生株及質體剔除株計算質體之備份數，pAH163 之備份數約為 500 左右，屬於高備份數之質體，而 pAH35 之質體備份數大約在 50~70 之間。另以將 pAH35、pAH163 之不同片段接於不含複製起始區之 E. coli 載體，建構含質體片段之重組質體，並以抗生素之抗藥性基因作為篩選標記以尋找複製起始區，但尚無明確結果。 | zh_TW |
| dc.description.abstract | Aeromonas hydrophila, a gram negative, facultatively anaerobic freshwater bacterium, occurring widely in aquatic environment, is an opportunistic pathogen of a variety of aquatic and terrestrial animals, including humans. As there is no suitable cloning system to study the gene function in Aeromonas spp. The purpose of this work is to construct vectors able to replicate in Aeromonas which contain replication origin from indigenous plasmids of Aeromonas hydrophila. After examining several clinical and environmental strains, six strains, A. hydrophila 25, A. hydrophila 35, A. hydrophila 132, A. hydrophila 163, A. hydrophila 191 and A. hydrophila 1028 carry plasmids were found. Plasmids pAH35 and pAH163 were isolated from A. hydrophila 35 and A.hydrophila 163 respectively and then were determined using WU-BlastN in EMBL database. Sequences similar to those of RNAII and RNAI were found in pAH35, suggesting pAH35 might be a ColE1-type plasmids. However, sequences similar to those of the RNAI and Rep protein of ColE2-type plasmids were also found in pAH35, suggesting pAH35 also might be a ColE2-type plasmid. Right now, the replication mechanism of pAH35 is unknown. In pAH163, we found a region encluding a replication related protein, Rep, which is needed in rolling circle replication. We used SDS, EtBr ,and higher incubation temperature in order to cure the plasmids of A. hydrophila 35 and A. hydrophila 163. One AH35 curing stain (AH35c1) and four curing stains of AH163 (AH163c1~4) were obtained. The biological character of the curing strains almost the same with the wild-type. We determined the copy number of the plasmids with wild-type and plasmid curing strains. The copy number of pAH163 is about 500 copies, therefore pAH163 belongs to high copy number plasmid. In the other hand, the copy number of pAH35 is about 50 to 70 copies. Using different fragment of pAH35 and pAH163 fused with E. coli vector to find the replication origin is still proceeding. | en |
| dc.description.provenance | Made available in DSpace on 2021-06-08T06:07:18Z (GMT). No. of bitstreams: 1 ntu-96-R93b47107-1.pdf: 3526269 bytes, checksum: e0b36c821db442a74602bdb519078cc9 (MD5) Previous issue date: 2007 | en |
| dc.description.tableofcontents | 誌謝 i
中文摘要 ii 英文摘要 iii 目錄 v 表目錄 x 圖目錄 xi 縮寫表 xiv 第一章 前言 1 1.1質體介紹 1 1.1.1細菌質體的一般性質及類型 1 1.1.1.1接合性與非接合性質體 1 1.1.1.2單一備份與多備份質體 2 1.1.1.3質體的不相容群 2 1.1.1.4廣與窄宿主域的質體 2 1.1.2質體的功能 3 1.1.2.1致育因子 3 1.1.2.2抗藥性質體 3 1.1.2.3產大腸桿菌素的質體 4 1.1.2.4分解利用性質體 4 1.1.2.5農桿菌的腫瘤誘導質體 4 1.1.3質體複製的調控 5 1.1.3.1質體的複製 5 1.1.3.2質體備份數之調控 6 1.2研究動機與目的 14 1.3實驗架構 14 第二章 材料 15 2.1菌株 15 2.2質體 15 2.3培養基 15 2.3.1 LB medium 15 2.3.2 LA medium 19 2.3.3 Skim milk palte 19 2.3.4 T.C.B.S. Cholera medium 19 2.4藥品 19 2.5儀器 21 第三章 研究方法 22 3.1細菌培養與測定 22 3.1.1生長曲線測定 22 3.1.2菌株特性分析 22 3.1.2.1 Skim milk plate assay 22 3.2質體剔除實驗 23 3.2.1剔除劑濃度選擇 23 3.2.2質體剔除－SDS、EtBr 23 3.2.3質體剔除－heat 24 3.3 DNA操作 24 3.3.1染色體 DNA 的抽取 24 3.3.2小規模的質體製備 26 3.3.3限制酶作用 28 3.3.4 DNA 片段的回收---酒精沉澱法 28 3.3.5 DNA 片段的回收---Viogene DNA extraction kit 29 3.3.6去除DNA鏈的 5' 端磷酸基 30 3.3.7接合反應 31 3.3.8細胞轉形 CaCl2 法與受容細胞的製備 32 3.3.8.1受容細胞 (competent cell) 的製備 32 3.3.8.2細胞轉形 CaCl2 法 33 3.3.9電轉形法 (electroporation) 與受容細胞的製備 33 3.3.9.1受容細胞的製備 33 3.3.9.2電轉形法 (electroporation) 34 3.3.10質體快速檢驗法 35 3.3.11 DNA 電泳 36 3.4蛋白質操作 37 3.4.1總蛋白質製備 37 3.4.2 Bradford 蛋白質定量法 38 3.4.3蛋白質膠體電泳 39 3.4.4膠片染色法及脫色法 41 第四章 結果與討論 43 4.1 Aeromonas hydrophila中質體之檢測 43 4.1.1質體檢測 43 4.1.2質體限制酶切位 45 4.2 Aeromonas hydrophila 內生質體之剔除 50 4.2.1質體剔除 (plasmid curring) 50 4.2.2 A. hydrophila 質體剔除株之生理特性 52 4.2.2.1質體與寄主細胞之生長 58 4.2.2.2質體與寄主細胞生長時期 58 4.2.2.3質體與胞外分解酵素 61 4.2.2.4質體與抗生物質抗性 61 4.2.2.5質體與胞內蛋白質 67 4.2.3質體備份數 (Plasmid copy number) 68 4.3 Aeromonas hydrophila中質體之序列分析 76 4.3.1核酸序列定序 (DNA sequencing) 76 4.3.2核酸序列分析 80 4.4質體複製區域之確認 96 第五章 結論 100 參考文獻 102 附錄 106 | |
| dc.language.iso | zh-TW | |
| dc.title | 嗜水性產氣單胞桿菌 Aeromonas hydrophila 質體 pAH163、pAH35 之性質分析 | zh_TW |
| dc.title | Characterization of plasmid pAH163 and pAH35 from Aeromonas hydrophila | en |
| dc.type | Thesis | |
| dc.date.schoolyear | 95-2 | |
| dc.description.degree | 碩士 | |
| dc.contributor.oralexamcommittee | 張敏政,黃健雄 | |
| dc.subject.keyword | 嗜水性產氣單胞桿菌,質體,質體剔除,質體備份數,定序, | zh_TW |
| dc.subject.keyword | Aeromonas hydrophila,plasmid,plasmid curing,copy number,DNA sequencing, | en |
| dc.relation.page | 141 | |
| dc.rights.note | 未授權 | |
| dc.date.accepted | 2007-07-19 | |
| dc.contributor.author-college | 生命科學院 | zh_TW |
| dc.contributor.author-dept | 微生物與生化學研究所 | zh_TW |
| 顯示於系所單位： | 微生物學科所 | |
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