葉怡均Yeh, Yi-Chun蔡思慈Tsai, Ssu-Tzu2023-12-082027-07-052023-12-082022https://etds.lib.ntnu.edu.tw/thesis/detail/bb130171ba8b44405a028149905fcd71/http://rportal.lib.ntnu.edu.tw/handle/20.500.12235/120901全細胞生物傳感器是一種將活細胞作為生物識別元件的生物傳感器，可用於檢測特定有毒物質或代謝物等，具有對環境友善、低成本和可攜帶性等優點。本論文主要基於邏輯閘概念去設計出不同的全細胞生物感測器：(1)在大腸桿菌中比較不同合成基因的汞離子感測器；(2)在耐金屬貪銅菌中開發銅金離子感測器。第一部分，根據邏輯閘概念，我們分別設計四種經過重組基因修飾的全細胞生物傳感器。在汞離子(Hg2+)和異丙基β-d-1-硫代半乳糖苷(IPTG)分子的控制下，分別比較了四對基因，包括LuxRI、HrpRS、Spy Tag/Catcher和E4/K4捲曲螺旋。此外，我們還研究了這些全細胞生物傳感器在各種pH水平下及其金屬干擾。第二部分，根據第一部份結果我們選擇成功表達的三個系統(LuxRI、HrpRS、Spy Tag/Catcher)，移到耐金屬貪銅菌(Cupriavidus metallidurans)中建立基於邏輯閘概念之銅及金離子感測器。Whole-cell biosensors are a type of biosensor that incorporates living cells as a biorecognition element, which can be used to detect specific toxic substances or metabolites. They have many advantages of being environmentally friendly, low-cost, and portable. This thesis mainly designs different whole-cell biosensors based on the concept of the logic gate: (1) Comparing mercury whole-cell biosensors combined with different synthetic genes in Escherichia coli; (2) Developing copper and gold ions whole-cell biosensors in Cupriavidus metallidurans.In the first part, according to the logic gate concept, we designed four whole-cell biosensors that were modified by recombinant genes, separately. Four pairs of genes were compared respectively including LuxRI, HrpRS, Spy Tag/Catcher, and E4/K4 coiled-coil under the control of mercury ions (Hg2+) and isopropyl β-d-1-thiogalactopyranoside (IPTG) molecules. Moreover, we also investigated these whole-cell biosensors under various pH levels and their metal interference.In the second part, according to the results of the first part, we selected three systems (LuxR/LuxI, HrpR/HrpRS, Spy Tag/Catcher) optimized from the first part, and expressed them in Cupriavidus metallidurans to detect copper and gold ions based on the logic gate concept.全細胞生物感測器重金屬調控系統邏輯閘重組蛋白耐金屬貪銅菌大腸桿菌銅離子汞離子金離子Whole-Cell BiosensorsMetalsRegulatory SystemsLogic GateRecombinant ProteinsCupriavidus metalliduransE. coliCopper ionsMercury ionsGold ionsetd