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Mini Photobioreactors for in Vivo Real-Time Characterization and Evolutionary Tuning of Bacterial Optogenetic Circuit
journal contribution
posted on 2017-05-22, 00:00 authored by Hsinkai Wang, Ya-Tang YangThe
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 in vivo real-time optogenetic
characterization of E. coli 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.