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| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 楊?伸(Chii-Shen Yang) | |
| dc.contributor.author | Ho Wang | en |
| dc.contributor.author | 王赫 | zh_TW |
| dc.date.accessioned | 2021-06-17T06:17:18Z | - |
| dc.date.available | 2019-08-23 | |
| dc.date.copyright | 2018-08-23 | |
| dc.date.issued | 2018 | |
| dc.date.submitted | 2018-08-21 | |
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The Rate of Osmotic Downshock Determines the Survival Probability of Bacterial Mechanosensitive Channel Mutants. Journal of Bacteriology 197, 231-237, (2015). | |
| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/71973 | - |
| dc.description.abstract | 長久以來，科學界著重研究動物細胞上的離子通道與幫浦，然而真核生物離子通道的結構與功能之成果，大多係建立於原核生物離子通道與幫浦研究基礎上。透過了解微生物離子通道與幫浦的生化與生理性質，將有助於更進一步分子層次上的探討。有別於常用於通道研究的非洲爪蟾 (Xenopus laevis) 卵母細胞，大腸桿菌 (Escherichia coli) 最常被用來表達和研究微生物視紫質 (microbial rhodopsin，MRho) 中一種感光蛋白質—光驅動離子幫浦之離子傳輸功能。但許多因素可能影響細胞膜，進而改變其上離子通道與幫浦之結構與功能；滲透壓便是其中一種。本研究於大腸桿菌表達不同物種的離子型微生物視紫質，並與大腸桿菌細胞本身，進行了於不同滲透壓環境下，全細胞光電流測試 (Whole cell photocurrent measurement)。結果發現，改變外部滲透壓，影響了表達於大腸桿菌上微生物視紫質的傳輸離子活性值。此外，上述之滲透壓改變，對表達不同種類的微生物視紫質 (光驅動離子幫浦) 的大腸桿菌，得到不同影響的活性值。另外透過光學顯微鏡觀察大腸桿菌在不同滲透壓環境下的細胞型態，藉以輔助和分析光電流測試結果之可靠性。本研究以新的角度，探討以大腸桿菌系統研究微生物視紫質的離子傳輸活性和所在環境之影響，未來，更可考慮藉由直接量測以此系統表達之目標蛋白質的離子電導訊號 (ion conductivity) 來了解光驅動離子的傳輸行為。 | zh_TW |
| dc.description.abstract | Studies of ion channels and pumps have been focused on animal-centric approach in the past, and indeed gained convincing achievements; while the progress of resolving structures and activities of eukaryotic ion channels are still mainly contributed by bacterial experimental models. Other than Xenopus oocytes, Escherichia coli is one of the often-adopted bacterium to express ion pumping microbial rhodopsin. Several factors, including osmotic pressure changes, could influence the physiology and structure of the membrane and further affect performance of expressed ion channels and pumps. This study investigates the ion transportation of ion-type microbial rhodopsins expressed in E. coli, Measuremtents of the light-driven photocurrents of microbial rhodopsins expressed on E. coli membrane under different osmotic environments were conducted. The results show that the external osmolarity affected the ion transportation behaviors of microbial rhodopsins expressed in E. coli. Such osmolarity changes affected differently at different expressed microbial rhodopsins. Observations of the E. coli morphological changes under different osmolarity environments supported the results of photocurrent measurements. We believe such approach provides new insights of using E. coli system to study ion transportation signals of microbial rhodopsins. Also, we propose directly measuring of the ion conductivity on the target protein expressed E. coli cells to investigate ion transportation activity. | en |
| dc.description.provenance | Made available in DSpace on 2021-06-17T06:17:18Z (GMT). No. of bitstreams: 1 ntu-107-R05b22043-1.pdf: 4829277 bytes, checksum: 97dc7de5b99240235f20819b75d65467 (MD5) Previous issue date: 2018 | en |
| dc.description.tableofcontents | 目錄 i
圖目錄 iv 表目錄 vi 摘要 vii Abstract viii 第一章 緒論 1 第一節 嗜鹽古生菌 1 第二節 微生物視紫質 (Microbial Rhodopsin) 2 I -2.1動物與微生物視紫質 2 I -2.2離子幫浦型視紫質 (Ion-pumping Microbial Rhodopsin) 3 I -2.3離子幫浦型視紫質的結構 4 第三節 微生物離子通道 7 I -3.1 微生物離子通道的種類與功能 7 I -3.2 離子通道與幫浦的應用 8 第四節 研究離子通道與幫浦的方法 9 I -4.1膜上離子通道之測量 9 I -4.2非洲爪蟾卵母細胞用於離子通道研究 11 第五節 滲透壓與離子傳輸之關聯 12 第二章 材料與方法 17 第一節 實驗材料 17 II -1.1菌種 17 II -1.2質體 17 II -1.3藥品 17 第二節 實驗儀器與設備 19 II -2.1核酸電泳 19 II -2.2離心機 19 II -2.3離子幫浦實驗用儀器 19 II -2.4光電流測量用儀器 19 II -2.5 細胞型態觀察用器材 20 II -2.6 電導度計 20 II -2.7其他 20 第三節 實驗方法 21 II -3.1 DNA建構 21 II -3.2光驅動離子幫浦活性測試 24 II -3.3全細胞光電流訊號量測 (Whole cell Photocurrent measurement) 25 II -3.4電導度量測 25 II -3.5大腸桿菌細胞型態觀察 25 第三章 結果與討論 27 第一節 大腸桿菌在不同滲透壓下的細胞形態 27 第二節 大腸桿菌於不同滲透壓下之光電流訊號 30 第三節 不同滲透壓下氫視紫質離子傳輸之影響 33 III -3.1 氫視紫質離子傳輸受外在滲透壓所影響 33 III -3.2 不同離子種類貢獻之滲透壓影響氫視紫質離子傳輸行為 34 第四節 不同滲透壓下氯視紫質離子傳輸之影響 35 III -4.1 氯視紫質之離子傳輸行為受外在滲透壓影響 35 III -4.2 無氯環境下氯視紫質離子傳輸行為 36 第五節 滲透壓對不同菌種之氯視紫紅質離子傳輸之影響 37 III -5.1 不同滲透壓下對 HwHR 之氯視紫紅質離子傳輸之影響 37 III -5.2 不同滲透壓下對 HmHR 之氯視紫紅質離子傳輸之影響 38 第六節 增加光刺激時間對離子幫浦型微生物視紫質離子傳輸之影響 39 III -6.1 氫視紫質在加長照光時間下的離子傳輸行為 39 III -6.2 氯視紫質在加長照光時間下的離子傳輸行為 40 第七節 光電流訊號量化分析 41 III -7.1 照光時段 41 III -7.2 後照光時段 42 第八節 微生物視紫質在不同滲透壓下之電導度量測 43 III -8.1 氫視紫質在不同滲透壓下受光刺激之電導度變化 45 III -8.2 氯視紫質在不同滲透壓下受光刺激之電導度變化 46 第九節 外在滲透壓與大腸桿菌系統離子傳輸關係之模型 48 第四章 結論 51 第五章 未來方向 53 第一節 細胞形態與滲透壓 53 第二節 大腸桿菌滲透壓調節系統研究 55 第三節 滲透壓即時改變與偵測 56 第四節 ITO 系統之優化 57 參考文獻 58 | |
| dc.language.iso | zh-TW | |
| dc.subject | 滲透壓 | zh_TW |
| dc.subject | 離子傳輸 | zh_TW |
| dc.subject | 離子通道與幫浦 | zh_TW |
| dc.subject | 微生物視紫質 | zh_TW |
| dc.subject | microbial rhodopsin | en |
| dc.subject | ion channels and pumps | en |
| dc.subject | ion transportation | en |
| dc.subject | osmotic pressure | en |
| dc.title | 於大腸桿菌探討微生物視紫質在不同滲透壓環境之光驅動離子傳輸 | zh_TW |
| dc.title | Probing Light-driven Ion Transportation of Microbial Rhodopsin under Different Osmotic Environments Using E. coli | en |
| dc.type | Thesis | |
| dc.date.schoolyear | 106-2 | |
| dc.description.degree | 碩士 | |
| dc.contributor.oralexamcommittee | 李昆達(Kung-Ta Lee),吳?承(Hsuan-Chen Wu),梁博煌(Po-Huang Liang),楊長豪(Chang-Hao Yang) | |
| dc.subject.keyword | 微生物視紫質,離子通道與幫浦,離子傳輸,滲透壓, | zh_TW |
| dc.subject.keyword | microbial rhodopsin,ion channels and pumps,ion transportation,osmotic pressure, | en |
| dc.relation.page | 61 | |
| dc.identifier.doi | 10.6342/NTU201804071 | |
| dc.rights.note | 有償授權 | |
| dc.date.accepted | 2018-08-21 | |
| dc.contributor.author-college | 生命科學院 | zh_TW |
| dc.contributor.author-dept | 生化科技學系 | zh_TW |
| 顯示於系所單位： | 生化科技學系 | |
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