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| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 李佳音 | |
| dc.contributor.author | Ya-Wen Chang | en |
| dc.contributor.author | 張雅雯 | zh_TW |
| dc.date.accessioned | 2021-06-16T17:21:41Z | - |
| dc.date.available | 2017-08-27 | |
| dc.date.copyright | 2012-08-27 | |
| dc.date.issued | 2012 | |
| dc.date.submitted | 2012-08-16 | |
| dc.identifier.citation | 張絜雯 (2009). 胞外絲胺酸蛋白酶對腸炎弧菌生物膜生成及遊走能力之影響. 國立台灣大學農業化學系碩士論文.
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| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/63876 | - |
| dc.description.abstract | 腸炎弧菌 ( Vibrio parahaemolyticus )以磷脂醯乙醇胺( Phosphatidylethanolamine, PE ) 為主要的膜磷脂質成分。PE合成路徑藉由磷脂醯絲胺酸合成酶 ( Phosphatodylserine synthase, Pss )以及磷脂醯絲胺酸脫羧酶 ( Phosphatidylserine decarboxylase, Psd )合成PE。本研究比較一株缺乏熱穩定性溶血素( tdh ) 以及類熱穩定性溶血素(trh )的腸炎弧菌臨床菌株，Vibrio parahaemolyticus No.93 ( VP93 ) 和此株腸炎弧菌pss突變株 ( Δpss )其生理變化現象，觀察剔除pss基因造成的影響。以薄層層析法分析兩菌株的磷脂質，發現Δpss –VP93的PE僅從野生株的72%降至63%，雖然Δpss –VP93有大量的PE殘存但仍有生長上的缺陷，包含在鹼性蛋白腖含鹽培養基(APSB)中生長速率下降了28%、在胰化蛋白大豆含鹽培養基(TSB3)中的菌體生成量只有野生株的一半、以及在固態培養基上的菌落較小。膜磷脂質成分的改變也影響菌體對於外來物質通透性的改變，像是對疏水性的抗生素氯黴素的抗性由最小抑制濃度10 μg/ mL上升至40 μg/ mL，同樣的疏水性抗生素紅黴素最小抑制濃度由5 μg/ mL上升至40 μg/ mL。以及對兩性離子界面活性劑CHAPS (3-[3-(膽醯胺丙基)二甲氨基]丙磺酸内鹽) 的耐受性增加、對陰離子界面活性劑SDS(十二烷基硫酸鈉) 的耐受性下降。腸炎弧菌pss突變株，Δpss –VP93生物膜的合成量與野生株VP93不同，無論在25℃、30℃ 或35℃下，TSB3或是APSB培養基中，其生物膜的合成量都比VP93高且具有顯著差異，但在APSB培養基中若額外添加20 mM MgCl2，則可抑制Δpss –VP93高生物膜合成量的此一表型。由以上結果得知腸炎弧菌在剔除pss基因後，造成膜磷脂質成份改變，影響了細胞的生長、細胞膜特性與生物膜的生合成。 | zh_TW |
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| dc.description.tableofcontents | 誌謝 ii
中文摘要 iv 英文摘要 v 目錄 vi 表次 ix 圖次 x 附錄圖次 xi 縮寫表 xii 壹、前言 1 第一節 腸炎弧菌 ( Vibrio parahaemolyticus ) 1 一、概論 1 二、性狀 1 三、毒力因子 2 第二節 膜脂質功能與成分 2 一、細胞膜脂質 2 二、細菌的膜磷脂質成分 3 三、二價金屬離子對細胞膜穩定性的幫助 4 四、細胞膜特性改變造成表型差異 4 第三節 磷脂醯絲胺酸合成酶與磷脂醯絲胺酸脫羧酶 5 一、路徑 5 二、磷脂醯絲胺酸合成酶的分類 6 三、原核生物中的磷脂醯乙醇胺及磷脂醯膽鹼的致病性功能 6 四、真核生物中的磷脂醯乙醇胺的致病性功能 7 五、磷脂醯乙醇胺及磷脂醯絲胺酸對真核細胞壁的結構的影響 7 第四節 生物膜 7 一、生物膜 7 二、生物膜形成 8 三、生物膜結構 9 第五節 細胞膜通透性 10 一、細胞膜組成份改變對抗性的影響 10 二、細胞膜組成份改變對膜特性的影響 10 三、細胞膜組成份改變菌體通透性 10 第六節、研究目的與源起 11 貳、實驗材料與方法 12 第一節 實驗材料 12 一、菌株、質體與引子 12 二、培養基 12 三、藥品試劑 12 四、商業套組 13 五、儀器 13 第二節 實驗方法 14 一、DNA技術 14 二、建構腸炎弧菌 psd突變株 18 三、建構腸炎弧菌 Δpss補償株 21 四、腸炎弧菌野生株與突變株生理特性分析 22 五、抗生素抗性測試 ( Minimum Inhibitory Concentration, MIC assay ) 23 六、膜特性分析 ( Cell Inhibition Assay ) 24 七、生物膜生合成 25 八、統計分析 ( Statistic Analysis ) 25 參、實驗結果 26 一、腸炎弧菌野生株與pss突變株生長曲線與速率 26 二、腸炎弧菌野生株與pss突變株菌落表型差異 26 三、腸炎弧菌野生株與pss突變株磷脂質成分分析 27 四、腸炎弧菌pss突變株對抗生素耐受性的改變 27 五、腸炎弧菌突變株對界面活性劑的耐受程度 28 六、腸炎弧菌野生株與pss突變株生物膜生合成能力 28 一、綜合比較 28 二、不同溫度下生物膜生合成 29 三、不同培養基下生物膜生合成 29 四、不同鎂離子濃度生物膜生合成 29 七、建構腸炎弧菌psd 缺陷株( Δpsd-VP93 ) 29 八、建構腸炎弧菌pss 補償株 ( Δpss-VP93/ Ppss ) 30 肆、討論 32 一、磷脂醯乙醇胺 ( PE ) 仍存在於腸炎弧菌pss突變株 32 二、腸炎弧菌pss突變株對氯黴素以及紅黴素抗性的提升 32 三、磷脂質組成的改變造成腸炎弧菌膜特性的改變 32 四、腸炎弧菌pss突變株具有較高的生物膜生合成能力 33 五、腸炎弧菌pss突變株不具有細胞分裂的缺陷 33 六、腸炎弧菌pss基因的角色 34 伍、結論 35 陸、參考文獻 36 | |
| dc.language.iso | zh-TW | |
| dc.subject | 腸炎弧菌 | zh_TW |
| dc.subject | 磷脂醯絲胺酸合成酶 | zh_TW |
| dc.subject | 生物膜 | zh_TW |
| dc.subject | biofilm | en |
| dc.subject | Vibrio parahaemolyticus | en |
| dc.subject | phosphatidylserine synthase | en |
| dc.title | 腸炎弧菌磷脂醯絲胺酸合成酶對細胞膜特性
與生物膜生合成的影響 | zh_TW |
| dc.title | The phosphatidylserine synthase affects the membrane property and biofilm formation
of Vibrio parahaemolyticus | en |
| dc.type | Thesis | |
| dc.date.schoolyear | 100-2 | |
| dc.description.degree | 碩士 | |
| dc.contributor.oralexamcommittee | 陳建先,余美萱 | |
| dc.subject.keyword | 腸炎弧菌,磷脂醯絲胺酸合成酶,生物膜, | zh_TW |
| dc.subject.keyword | Vibrio parahaemolyticus,phosphatidylserine synthase,biofilm, | en |
| dc.relation.page | 77 | |
| dc.rights.note | 有償授權 | |
| dc.date.accepted | 2012-08-17 | |
| dc.contributor.author-college | 生物資源暨農學院 | zh_TW |
| dc.contributor.author-dept | 農業化學研究所 | zh_TW |
| 顯示於系所單位： | 農業化學系 | |
文件中的檔案：
| 檔案 | 大小 | 格式 | |
|---|---|---|---|
| ntu-101-1.pdf 未授權公開取用 | 1.58 MB | Adobe PDF |
系統中的文件，除了特別指名其著作權條款之外，均受到著作權保護，並且保留所有的權利。