Speaker: Prof. Jack Ya-Tang Yang
Affiliation: Department of Electrical Engineering, National Tsing Hua Univerisity, Hsinchu, Taiwan
Gene circuits have been dynamically characterized by using chemical effectors molecules, which has been limited by diffusion of effector across the cellular membrane. Optical method for creating on-demand protein signals in live cells would bypass limitations and, in principle, enable the dynamical characterization of virtually any gene circuit that responds to changes in protein concentration. Current standard protocol to characterize such optogenetic circuit using flow cytometer is tedious, labor intensive and cumbersome. In this work, we will go over the bioreactor development in my group and such efforts have culminated in a bioreactor for optogenetic characterization of light sensing E. coli. We develop an integrated bioreactor that uses optogenetic control for quantitative, up/down control of gene expression and monitors the measurement of the relevant parameters during the microbial growth. The optical density, fluorescence detection for green fluorescence protein reporter as well as the input stimuli is provided by light emitting diode(LED) at multiplexed wavelength. The light sensing E. Coli harbors a synthetic two component system(TCS) circuit from CcaSCcaR system used in the chromatic adaption of cyanobacteria synechocystis PCC 6803. We have also characterized evolutionary stability of such TCS by serial dilution transfer. Such a tool will be vital for bacterial optogenetic circuit characterization.