Mini Photobioreactors for <i>in Vivo</i> Real-Time Characterization and Evolutionary Tuning of Bacterial Optogenetic Circuit
Published on 2018-02-10T00:04:40Z (GMT) by
The current standard protocols for characterizing the optogenetic circuit of bacterial cells using flow cytometry in light tubes and light exposure of culture plates are tedious, labor-intensive, and cumbersome. In this work, we engineer a bioreactor with working volume of ∼10 mL for <i>in vivo</i> real-time optogenetic characterization of <i>E. coli</i> with a CcaS–CcaR light-sensing system. In the bioreactor, optical density measurements, reporter protein fluorescence detection, and light input stimuli are provided by four light-emitting diode sources and two photodetectors. Once calibrated, the device can cultivate microbial cells and record their growth and gene expression without human intervention. We measure gene expression during cell growth with different organic substrates (glucose, succinate, acetate, pyruvate) as carbon sources in minimal medium and demonstrate evolutionary tuning of the optogenetic circuit by serial dilution passages.
Cite this collection
Wang, Hsinkai; Yang, Ya-Tang (2018): Mini Photobioreactors for in Vivo Real-Time Characterization
and Evolutionary Tuning of Bacterial Optogenetic Circuit. ACS Publications.