廖秀娟2006-07-252018-06-292006-07-252018-06-2920032003-07-31http://ntur.lib.ntu.edu.tw//handle/246246/10741生物偵測器（biosensors）是指sensing component (生化或生理的變化) 和transducing element (當環境改變時，會產生可測量的訊號) 之結合。對於基因改造（genetically engineered） 細胞的生物偵測器，可基因重組sensing element 和reporter gene，此重組的質體(recombinant plasmid)，會對外來污染物反應產生一種可量測的訊號。Sensing element 通常是由regulatory protein 和promoter 所組成，這些sensing element 對污染物的專一性決定此生物偵測器的選擇 性，而reporter protein 則是決定此生物偵測器的敏感性及偵測極限。 本研究乃研發以綠色螢光蛋白（green fluorescent protein，GFP）作為reporter protein 的專 一性有機污染物的生物偵測系統。針對環境中重要的有機污染物，例如：benzene 及其衍生物， 發展具專一性有機污染物的生物偵測系統。此種以GFP 作為reporter protein 的生物偵測器， 其優點是它具有自動發出螢光(auto-fluorescence)的特性，其發光反應不需要添加任何cofactors 或受質，因此，此種生物偵測器可提供迅速甚至即時（real time）環境中有機污染物資訊，此 種以GFP 作為reporter protein 生物偵測系統，將可克服luc，lux，及lacZ 作為reporter gene 的缺點。 本計畫已成功的建立適合原核生物之GFP-based 的質體，此質體將做為有機污染物生物 偵測器之vector。研究結果顯示此質體不具背景螢光，並具有高度之穩定性。另外建構benzene 生物偵測器之sensing elements 之一的promoter 已成功的分離。A biosensor is composed of a biological sensing component, which recognizes chemical or physical changes, couple to a transducing element that produces a measurable signal in response to the environmental insult. Genetically engineered cell-based biosensors can elicit a response in the presence of an environmental insult by coupling the sensing element to a reporter gene fusion, which upon expression produces a readily measurable signal. The sensing element is often composed of regulatory proteins and promoter. The specificity of these sensing elements for the environmental insult confers selectivity to the system, while the reporter protein determines the systems’s sensitivity and detection limits. This project studied the construction, laboratory characterization, and environmental testing of genetically engineered GFP-based bacterial biosensors that detect specific organic pollutants (e.g., benzene). The reporter plasmid in which expression of gfp gene for jellyfish Aequorea victoria green fluorescence protein (GFP) will be placed directly under the control of the specific promoter. The primary advantage of GFP as a reporter protein is its autofluorescence, and therefore, its use does not require the addition of cofactors or exogenous substracts to produce fluorescence. Therefore, such GFP-based biosensors might provide even real time monitoring environmental pollutants. We have successfully constructed a prokaryotic GFP-based vector. This vector is used for developing the benzene biosensor. The results indicated that the GFP-based vector has almost no fluorescence background and high stability. In addition, we have successfully isolated and amplified one of the promoter fragments that are essential for establishing the benzene biosensor.application/pdf179902 bytesapplication/pdfzh-TW國立臺灣大學生物環境系統工程學系暨研究所生物偵測器（biosensor）綠色螢光蛋白（GFP）reporter protein有機污染物biosensorsgreen fluorescence protein (GFP)organic pollutants[SDGs]SDG11reporthttp://ntur.lib.ntu.edu.tw/bitstream/246246/10741/1/912211E002096.pdf