TY - JOUR T1 - Prototyping of <em>Bacillus megaterium</em> genetic elements through automated cell-free characterization and Bayesian modelling JF - bioRxiv DO - 10.1101/071100 SP - 071100 AU - Simon J. Moore AU - James T. MacDonald AU - Sarah Weinecke AU - Nicolas Kylilis AU - Karen M. Polizzi AU - Rebekka Biedendieck AU - Paul S. Freemont Y1 - 2016/01/01 UR - http://biorxiv.org/content/early/2016/08/23/071100.abstract N2 - 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 the system, we have applied a Bayesian statistical approach to infer model parameters by simultaneously using information from multiple experimental conditions. These developments now open up a new approach for the rapid and accurate characterization of genetic circuitry using cell-free reactions from unusual microbial cell chasses for bespoke applications. ER -