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微型光生物反應器應用於活體細胞的即時表現與細菌光基因電路演化調控
Thesis

微型光生物反應器應用於活體細胞的即時表現與細菌光基因電路演化調控

王信凱
Masters, 國立清華大學, 電子工程研究所
2016

Abstract

生物反應器 演化 自動化 基因電路 螢光偵測 bioreactor evolve automated gene circuit fluorescent detection
Current standard protocol to characterize such optogenetic circuit of bacterial cells using flow cytometry in light tubes and well plates for light exposure is tedious, labor intensive and cumbersome. In this work, we engineer a bioreactor of working volume ~10 ml for in vivo, real time optogenetic characterization of E. coli with a CCaS-CCaR light sensing system. The optical density, fluorescence detection for reporter protein as well as the light input stimuli are provided by four light emitting diode sources and two photodetectors. Once calibrated, the device can cultivate the cells and record both the microbial growth and gene expression without human intervention. We measure gene expression for cell growth under different organic substrates(glucose, succinate, acetate, and pyruvate) as carbon source in minimal medium and demonstrate evolutionary tuning of optogenetic circuit by serial dilution passage.

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