Prototyping of Bacillus megaterium genetic elements through automated cell-free characterization and Bayesian modelling

Automation and factorial experimental design together with cell-free in vitro transcription-translation systems offers a new route to the precise characterization of regulatory components. This now presents a new opportunity to illuminate the genetic circuitry from arcane microbial chassis, which are difficult to assess in vivo. One such host, Bacillus megaterium, is a giant microbe with industrial potential as a producer of recombinant proteins at gram per litre scale. Herein, we establish a B. megaterium cell-free platform and characterize a refactored xylose-repressor circuit using acoustic liquid handling robotics to simultaneously monitor 324 reactions in vitro. To accurately describe t